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Determination of serum or plasma progesterone (P4) concentrations is important to recognize pregnant and non-pregnant ewes, and also to predict the number of carried lambs. The 2 most common methodologies for the detection of plasma P4 are radioimmunoassay (RIA) and enzyme immunoassay (EIA). RIA is very expensive, and not all laboratories are equipped to perform this test; EIA is commercially available for human use, but only a few companies produce species-specific kits, which are expensive. We verified for ovine plasma a less expensive and easily available ELISA kit (DiaMetra) designed to quantify P4 in humans. Pools of ovine and human plasma were used to compare repeatability, accuracy, sensitivity, and stability of P4 measured by the DiaMetra kit. Repeatability data were within 15%, and accuracy values were ~90% for both plasma matrices. Stability data showed a loss of

Vaccination was the main approach to control spreading of FMD disease in Egypt. This study was carried out for evaluation of vaccination by governmental and prevailing local vaccines in cattle sera in menofia governorate during the period from October 2018 to May 2019. A total 130 serum samples were collected to evaluate the antibodies against FMDV. 30 samples were used as control, 50 samples collected from animals received local governmental vaccine and 50 samples from animals received local prevailing vaccine. 49 out of 100 serum samples (37.7%) were positive and 81 out of 130 serum samples (62.3%) were negative for NSP. Results were (0.0), 3(3.7%) pre-vaccination and 27(33.33%), 18(22.22%) 1st MPV- 25(30.86%), 18(22.22%) 2nd MPV-15(18.52%), 13(16.05%) 3rd MPV and (0.0), 9(11.11%) 4th MPV for serotype A. For serotype (O): 9(11.1%), 10(12.4%) pre-vaccination- 36(44.44%), 28(34.56%) 1st MPV-33(40.74%), 27 (33.33%) 2nd MPV-24(29.63%), 20(24.7%) 3rd MPV-14(17.3%), 12(14.80%) 4th MPV. For serotype (SAT2): 4(4.9%), 24(4.93%) pre-vaccination-30 (37.03%), 21(25.93%) 1st MPV-27(33.33%), 21(25.93%) 2nd MPV- 18 (22.22%), 14(17.3%) 3rd MPV-3 (3.71%), 4(4.93%) 4th MPV. In Conclusion. FMD controlling requires vaccines that will supply disparity of infected from vaccinated animals.

Background: Echinococcosis caused by Echinococcus granulosus is a zoonotic disease of public health significance, but there have been few studies of the infection in dogs in Nigeria. This study aimed to establish the seroprevalence of E. granulosus in dogs in four Local Government Area of Plateau State, Nigeria.Methods: A total of 179 dog sera were examined for the presence of E. granulosus antibody using an ELISA kit between May to October 2017.Results: Eleven of the sera (6.1%) were seropositive with a prevalence of 7.4% and 2.3% for dogs less than 2 years and those above 2 years of age respectively. More male dogs (7.5%) than females (4.1%) were seropositive. Seroprevalence was associated with 2 years dogs (OR=3.33, p=0.46) and male than female dogs (OR=1.90, p=0.53) but association were not significant. Dogs from Bassa had the highest seroprevalence (15.2%), followed by Mangu (5.9%), Kanke (4.8%) and Jos South (2.0%). Odds ratio showed an association which was not significant between Bassa (OR=3.3), Kanke (OR=2.50).Conclusion: This study shows that E. granulosus infection is present in dogs in some parts of Plateau State. More studies should be done to ascertain echinococcus infection in dogs in Nigeria to aid the formulation of control programmes to forestall its public health impacts. Annals of Medical Laboratory Science (2021) 1(2), 9 - 17

Tumour protein 53 (p53) plays an important role in the instruction of the cell cycle. In a variety of transformed cell lines, tumour protein is expressed in high amounts, and it is believed to contribute to transformation and malignancy. This research aimed to detect the anti-p53 antibodies in sera of patients with various malignant tumours and to evaluate the sensitivity and specificity of an enzyme-linked immunosorbent assay (ELISA). A case-control study was conducted on samples from 49 patients with various types of malignant tumours at Sultanah Bahiyah Hospital, Alor Setar, Kedah, Malaysia, and 32 healthy control cases with non‐malignant disease collected from Universiti Sains Malaysia clinic, Penang, Malaysia. The antibodies against p53 protein in the serum samples were analysed using the commercial ELISA kit, Calbiochem® p53- ELISAPLUS. The results showed that the rate of anti-p53 antibodies in patients with various malignant tumours was 13 out of 49 (26.5 %), compared with only 2 out of 32 (6.25%) in healthy controls (p < 0.001). The sensitivity of this kit reached 28.6% and the specificity was 93.8%. In conclusion, these results suggest that the anti-p53 antibodies can be detected in different sera of malignant tumour patients and the ELISA kit is highly specific; nevertheless, its discrimination power is not perfect because of its low sensitivity to determine the anti-p53 antibodies.

This study was designed to detect the persistent infection with bovine viral diarrhea virus (PI- BVDV) among 114 bulls in Aswan Governorate. Sera samples were examined by ELISA and real time polymerase chain reaction (PCR) to detect BVDV antibodies and BVDV nucleic acid, respectively. The percent of infection was 23.7% (27/ 114) and 76.3% (87/ 114) were negative by ELISA technique, and 3.4% (3/ 87) for BVDV gene detection by real time PCR. Sensitivity of 89%and specificity of 96 % was reported using the commercially available ELISA kit for samples obtained from PI animals.

Mycoplasma gallisepticum (M. gallisepticum) is the primary agent of chronic respiratory disease causing important economic losses in the poultry industry. Compared to antibodies, aptamers used to diagnose M. gallisepticum have many advantages, such as being chemically, animal-free produced and easily modifiable without affecting their affinity. Herein, a single-stranded DNA (ssDNA) aptamer Apt-236 which can specifically bind to PvpA protein of M. gallisepticum with a Kd of 1.30 ± 0.18 nM was selected successfully. An indirect blocking ELAA (ib-ELAA) for M. gallisepticum antibodies detection was also developed using Apt-236, in which M. gallisepticum antibodies would block the binding-position of aptamers. Therefor positive sera would prevent color development whereas negative sera will allow a strong color reaction. The ib-ELAA was consistent with other three widely used assays in terms of the growth and decline of the antibody response to M. gallisepticum, and showed substantial agreement with the results obtained using a commercial ELISA kit in clinical chicken sera samples. Therefore, the ib-ELAA developed in this study was a new format for aptamer application and would be an alternative method for the surveillance of M. gallisepticum.

The cross-sectional seroprevalence study of the peste des petits ruminants (PPR) in sheep and goats was carried out in the Southern Peninsular region of India to ascertain the prevalence of PPR virus (PPRV) antibodies at the epidemiological units (epi-units) level in the small ruminant population. The serum samples were collected from various epi-units (villages) in the different states and union territory (UT) in Southern Peninsular region using a stratified random sampling methodology from August 2017 to March 2018. A total of 6643 serum samples [sheep (n = 2785) and goats (n = 3858)] were collected from 360 epi-units and were screened by PPR competitive ELISA kit for the detection of PPRV antibodies. The results revealed that the seroprevalence of PPR in small ruminants in Telangana, Andhra Pradesh, Karnataka, Tamil Nadu, and Kerala states, and Puducherry UT was 87.0%, 66.4%, 64.3%, 47.8%, 11.4%, and 50.4%, respectively in the studied region. Further, the results of the chi-squared test revealed that the PPRV antibodies across different states and UT in the region were associated (sheep-χ(2) = 218.8, p

El objetivo fue evaluar la frecuencia serológica individual y de rebaño, así como detectar los factores de riesgo de C. abortus, en siete estados donde se realiza la producción ovina en México. Se efectuó un estudio multifactorial, transversal y estratificado, con análisis de 5,321 muestras serológicas en 323 rebaños de 61 municipios. Se realizó ELISA con un kit comercial. Los factores de riesgo asociados con la enfermedad se determinaron mediante cuestionarios y análisis estadísticos con una prueba de Ji cuadrada con un intervalo de confianza del 95%. Del total de 5, 231 muestras de suero que fueron colectadas, 581 (10.92 %) dieron positivas a la prueba de ELISA. Los resultados por estado de sueros positivos fueron: Tlaxcala 13.08% (73/558); Sonora 12.45 % (102/819); Chihuahua 11.56 % (107/925); Hidalgo 11.34 % (97/855); Chiapas 10.15 % (60/591); Querétaro 9.69 % (79/815); Estado de México 7.09 % (63/758). La frecuencia de hatos seropositivos, fue de 43.34 % (140/323). Los resultados al ser agrupados por estado fueron los siguientes: Hidalgo 67.39 % (31/46), Querétaro 67.18 % (43/64); Sonora 40.92 % (19/47); Tlaxcala 33.33 % (12/36); Chiapas 31.57 % (12/38); Estado de México 25.45 % (14/55) y Chihuahua 24.32 % (9/37). Los principales factores de riesgo que favorecen la presencia del aborto enzoótico de las ovejas, fueron que estuviesen gestantes, que tuviesen una edad entre los 37 a los 48 meses y que el sistema de producción fuese intensivo. Estos estudios serológicos identificaron la presencia del aborto enzoótico de las ovejas en México, así como algunos de los factores de riesgo que inciden para favorecer su presencia.

Paratuberculosis (Ptb) is a widespread chronic infection caused by Mycobacterium avium subsp. paratuberculosis (Map) that affects both domestic and wild ruminants. Most of the studies focused on cattle while, the prevalence of Ptb in sheep in different regions of the world is not well investigated. This study aimed to address this gap of knowledge by screening adult female sheep for paratuberculosis antibodies in different geographical regions of Tunisia. A total number of 338 female sheep from 15 small to middle‐sized, extensively managed sheep farms in six regions across Tunisia were sampled. Animals were clinically examined before blood sampling. Sera were tested for the presence of anti‐Mycobacterium avium subsp. paratuberculosis antibodies using a commercial ELISA kit. Six farms out of 15 comprised at least one seropositive animal and 11 female sheep out of 338 tested animals (3.25%; 95% CI = [1.83–5.73]) were seropositive to M. avium subsp. paratuberculosis. The seroprevalence was significantly lower in 5‐year‐old females (p = .04) and animals that do not graze (p = .02). Due to its huge economic and social impacts, paratuberculosis represents a health problem in Tunisia and several other countries Further investigations are needed to rank sheep Ptb in Tunisia among other diseases and to assess the main risk factors using a larger nation‐wide survey., Six farms out of 15 comprised at least one seropositive animal and 11 female sheep out of 338 tested animals (3.25%; 95% CI = [1.83–5.73]) were seropositive to M. avium subsp. paratuberculosis, The seroprevalence was significantly lower in 5‐year old and older females (p = .04) and animals that do not graze (p = .02). This findings confirm that Paratuberculosis is circulating in sheep herds in Tunisia.

We examined anti-SARS-CoV-2 IgG and IgM antibodies in 45 serum samples from 26 patients with COVID-19, who were admitted in our hospital by using three different ELISA kits. All patients had pneumonia at admission, and 7 patients required mechanical ventilator support and grouped in severe case. Anti-SARS-CoV-2 IgG and IgM antibodies turned to be partially positive between the 6th and 10th days, more than 84% positive between the 11th and 15th days, and 100% after the 16th day. One ELISA kit revealed poorer sensitivity for anti-SARS-CoV-2 IgM antibody. Negative conversion of IgM antibody was not observed in the 30th day in our cohort. All three ELISA kits showed no false positive reaction for negative serum samples. Between severe and moderate cases, there was no significant difference in the trends of anti-SARS-CoV-2 IgG and IgM antibody.

The human T-cell lymphotropic virus types 1 and 2 (HTLV 1/2) are well known causes of adult T cell leukemia lymphoma. Both viruses were established to be transmitted through various mode including sexual contact and blood transfusion. This study was aimed to determine the seroprevalence of HTLV-1/2 antibodies among blood donors in Public Health Laboratory. During the period of August 2019 to October 2019 a total of 394 blood samples were obtained from blood donors visiting the Blood bank of National Public Health Laboratory, both males and females were included. The blood samples were analyzed for the presence of anti - HTLV-1,2 by a commercially available enzyme-linked immune-sorbent assay following the instructions of the manufacturer. The study participants were included 361 (91.6%) males and 33 (8.4%) females. The result show that of the 394 blood donors, four (4) were found to be seropositive for HTLV-1 antibodies giving a prevalence of 1.02%. While all samples were negative for the HTLV-2 antibodies. Among HTLV-1 positive cases 3 were male (75%) while only one female (25%) was found to be seropositive for HTLV-1 antibodies. None of the married donors was found to be seropositive for HTLV-1. We conclude that the seroprevalence of HTLV 1/2 were matched to the internationally estimated prevalence among blood donors at Blood Bank of national health laboratory and the majority of the cases were male under 40 years old. Further studies should be done with inclusion of more samples and using more sensitive technique like Western blot or PCR.

A cross sectional study was conducted during the period 2014-2015, to estimate the prevalence and assess contribution of some risk factors for the occurrence of enterotoxaemia in sheep, goats, cattle and camels in the Kingdom Saudi Arabia. A total of 1593 animals from 476 herds were investigated. ELISA tests were conducted using a commercial kit for the detection of different toxins produced by Clostridium perfringens causing the disease. The toxicogenic typing of C. perfringens in clinical samples by ELISA kit, revealed that the predominant types were C. perfringens type A (67.2%) followed by type D (16.4%), then type B (13.4%) and type C (3%). Further confirmation of representative C. perfringens isolated from enterotoxaemia positive animals was performed using polymerase chain reaction. The overall prevalence of enterotoxaemia disease at the animal and herd level was 27.2% and 26.47% respectively. There were significant differences in the prevalence of enterotoxaemia in animals between the different regions in KSA, animal species and months of the year. The highest prevalence was detected in Aljouf region (41.7 %), followed by Hail region (40.9%), Qassim (37.8%), Jazan (31.1%), the eastern region (26.1%) and Riyadh region (10.4%). The prevalence was the highest cattle (64.3%), followed by goats (29.9%), camels (21.5%) and sheep (21.4%). Risk factors such as regions (χ2 = 89.65, p = 0.000), months of the year (χ2 = 76.65, p = 0.000), animal species (χ2 = 50.81, p = 0.000), disease presence by years (χ2 = 29.75, p = 0.000), herds prevalence (χ2 = 1443.6, p = 0.000) showed statistically significant association with enterotoxaemia disease. In the multivariate analysis, species of animals (OR = 4.88, p = 0.000) was found to be the most statistically significant risk factor associated with enterotoxaemia. Cattle were found to be approximately five times more likely to have infection with enterotoxaemia (OR = 4.88, CI = 2.55-9.37, p=0.0001) compared to other animal species. High overall prevalence of enterotoxaemia cases in all animal species occurred during the period August – December of the year 2015 with the highest prevalence in September (50%).

Objective To explore whether auricular concha eletroacupuncture (ACEA) is effective in regulating lipid-lipoprotein metabolism in rats submitted to cold stress. Methods Thirty-six adult male Sprague–Dawley rats were placed in four groups(9 rats in each group), the rats in three groups of which were submitted to cold stress for fourteen days, the last one of which was a control group. After the cold stress process, in those three groups, the rats of one group were with no treatment (stress only), two were treated with either ACEA or auricular margin eletroacupuncture (AMEA) repeated for fourteen days. On the 14th day, all the rats were sacrificed after all experimental procedure for blood sampling. Blood glucose, triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) levels were tested by using the collected serum. Plasma lecithin-cholesterol acyltransferase (LCAT) was measured with ELISA kit. Results ACEA down-regulated the TG level (P Conclusion ACEA played an important role in regulating lipid-lipoprotein metabolism in rats submitted to cold stress.

Consumption of game in Germany has increased during the past 10 years. Wild boar (Sus scrofa), roe deer (Capreolus capreolus) and red deer (Cervus elaphus) are the most frequently hunted and consumed game animals in Germany, yet information on the occurrence of zoonotic pathogens in these animal species is scarce. To better estimate the public health risk emanating from handling and consumption of game, this study investigated seroprevalences of Toxoplasma gondii in game hunted in the German federal state of Brandenburg during two hunting seasons from 2017 to 2019. Toxoplasma gondii-specific antibodies were detected in 24.4% (44/180, 95% CI: 18.4%-31.4%) of wild boar, 12.8% (16/125, 95% CI: 7.5%-20%) of roe deer and 6.4% (3/47, 95% CI: 1.3%-17.5%) of red deer using a commercial ELISA kit. Seroprevalences were similar in the two hunting seasons. Correlation between sex and seropositivity could not be observed. A rise in seroprevalence was seen with increasing age in all studied game species. Observed seroprevalences suggest that T. gondii is endemic in the sylvatic environment in the German federal state of Brandenburg and imply that game could represent a relevant source for human T. gondii infection.

Introduction: Cryptococcus neoformans is an encapsulated fungal pathogen that causes life-threatening meningoencephalitis in immunocompromised patients. This yeast secretes several potent immunogenic proteins by secretory vesicular mechanisms, such as HSP70 chaperone. Methods: The PCR-amplified HSP70 gene was cloned into a PET-28a(+) expression vector. The purified recombinant HSP70 (rHSP70) was evaluated by western blotting using an anti-His Tag-HRP antibody and then used for immunization of a rabbit. The serum of the immunized rabbit was tested against the whole lysate of C. neoformans in ELISA. Results: The antibodies in the rabbit’s serum recognized lysate of C. neoformans yeast. The highest antibody levels were achieved after the third booster injection. Conclusion: The rHSP70 showed to be a reliable candidate for the designing and development of an ELISA kit for early detection of cryptococcosis, and to screen a large number of specimens.

Sepsis is a cause of non-cardiac death in the hospital. Early and rapid diagnosis of septic patients is a challenge toincrease the expectancy of life. IL-6 and TNF-α are groups of pro inflammatory cytokines that initiate an initial inflammatoryresponse. Procalcitonin is a specific marker of bacterial infection. This study aimed to analyze the correlation of serumcytokine IL-6, TNF-α, procalcitonin and leukocyte count in suspected sepsis patients. This was a cross-sectionalobservational study consisting of 45 patients with suspected sepsis with procalcitonin level > 0.5 ng/mL. Procalcitonin levelwas measured with Enzyme-Linked Fluorescent Assay (ELFA) (VIDAS), IL-6 and TNF-α levels were measured with theU-CyTech Human Elisa kit (Bioscience, INC) and leukocyte counts were measured with SYSMEX-XN 1000. It was found in thisstudy that IL-6 levels ranged in 0 pg/mL – 73.29 ng/mL (mean 29.43 ng/mL), TNF-α levels were 0 pg/mL – 390.5 pg/mL (mean27.62 pg/mL), and the mean value of leukocytes was 20,139/μL. There was no correlation between leukocyte counts with IL-6(p=0.798 and r=0.040), TNF-α (p=0.304 and r = -0.160), and procalcitonin (p=0.323 and r = 0.154). There was no correlationbetween IL-6 levels with TNF-α levels (p=0.871 and r = -0.025), and procalcitonin levels (p = 0.466 and r = 0.112). There wasa weak negative correlation between TNF-α level and procalcitonin levels (p=0.006 and r = -0.403) and there was a weaknegative correlation between procalcitonin and TNF-α levels in suspected sepsis patients.

Objective The objective of the research was to determine the giving of Holothuria scabra on the level of Docoxahexaenoic Acid (DHA) of breast milk with preterm chilbirth. Methods The research was a quasi experimental study with control group design. The samples were postpartum mothers with preterm childbirth consisted of 40 people divided into two groups. Postpartum mothers given Holothuria scabra capsule were intervention group and the postpartum mothers not given Holothuria scabra were control group. The sample of mother's milk of pretest of each group was taken in the seventh day and seven day later the sample was taken 3 cc. Intervention was done for 7 days per sample with 3 times dosage consisted of 2 capsules per day. The sample of breast milk was examined with Human DHA ELISA kit. The data were analyzed using Mann–Whitney test and Wilcoxon test. Results The results of the research indicated the difference of DHA level between intervention group and control group with a value of p = value 0.006. After the intervention, the average increase was 187.02 μg/mL for intervention group and 7.05 μg/mL for control group with a value of 1151.04 μg/mL.

Allergic responses are mainly caused by IgE, which is often located on the cell surface. The current diagnostic method detects both allergen-specific IgE and total IgE levels, but a number of allergic patients have a normal serum IgE level, which is a poor clinical correlate for allergy. Here, we developed a simple method to detect the level of cell-bound IgE by dissociating it from blood cells with lactic acid. Dissociated cell-bound IgE and plasma IgE levels were detected using the same ELISA kit at the same time. We established two clinical cohorts: an allergic patient group and a healthy participant group. In general, cell-bound IgE correlated well with plasma IgE; however, some patients exhibited high cell-bound IgE levels but low plasma IgE levels. We recommended 350 ng/mL peripheral blood total IgE (cell-bound IgE + plasma IgE) as the cut-off value for allergy diagnosis. Using this indicator, 90.32% of our allergic patients were correctly diagnosed. The peripheral blood total IgE level is a promising clinical diagnostic indicator in allergic patients and will provide more guidance for allergy diagnosis and therapeutic evaluation.

Rift Valley fever (RVF) is an emerging mosquito-borne zoonosis that threatens public health and animal agriculture in the endemic areas causing devastating epizootics characterized by abortion storms and high mortalities, especially in newborn animals. A cross-sectional study was conducted to determine the seroprevalence and investigate risk factors associated with exposure to the virus in camels slaughtered in Maiduguri abattoir, Borno state of Nigeria. Camels presented for slaughtered were sampled and data on age, sex, source or origin, utility, presence of post mortem lesions, body weights and body condition score were collected. Blood samples were collected and sera were harvested and stored at -20°C until tested. The sera were tested using a commercial ELISA kit based on the manufacturer's instructions. The overall seroprevalence in the study was 20.7% (95% CI, 13.6 – 30.0). The analysis showed no significant differences between presence of antibodies and variables that included the sex of camels (c2=0.015, df = 1, p = 0.904) and presence of post mortem lesion on the carcass (c2=0.009, df = 1, p = 0.925). There were significant differences between presence of antibodies and three variables that included the age (c2= 4.89, df = 1, p = 0.027), the source (c2= 7.077, df = 2, p = 0.029) and the main utility (c2= 8.057, df = 3, p = 0.045) of the camels. It was concluded that camels presented for slaughter at the Maiduguri abattoir have evidence of exposure to the RVF virus and may be means of transmission of the virus. Regular monitoring and control of transboundary animal movements were recommended in the study area.

Background: An apicomplexan protozoon Neospora caninum, causative agent of neosporosis, is recognized as one of the most common and important cause of sporadic and endemic bovine abortion and reduced reproductivity in dairy and beef cattle worldwide. The present study was conducted to investigate the relationship between N. caninum seroprevalence and infertility problems in 400 cows in Burdur city, Turkey. Methods: Blood samples were collected from vena jugularis into sterile serum tubes from 49 aborted, 58 infertil, 48 pregnant and 245 healthy cows for the findings of reproductive anamnesis during a period of March 2010 to March 2011. Sera samples were analyzed by competitive ELISA kit. Results: The seroprevalences were 7.7%, 6.4% and 4.2% in 2-4, ≤2 and ≥4 age groups respectively and no statistically significance observed between age groups. Seropositivity rates were 5.7%, 5.1%, 4.5%, 3.6% in Holstein, Montofon, cross-breeds and Simental breeds respectively. Seroprevalence differences was not statistically significant among cattle breeds. Antibodies to N. caninum were found in rates of 16.3%, 6.9%, 6.3%, 2.4% in aborted, infertile, pregnant and healthy cows respectively and there was a significant difference (P

Multiple serotypes and topotypes of foot-and-mouth disease virus (FMDV) circulate in endemic areas, posing considerable impacts locally. In addition, introductions into new areas are of great concern. Indeed, in recent years, multiple FMDV outbreaks, caused by topotypes that have escaped from their original areas, have been recorded in various parts of the world. In both cases, rapid and accurate diagnosis, including the identification of the serotype and topotype causing the given outbreaks, plays an important role in the implementation of the most effective and appropriate measures to control the spread of the disease. In the present study, we describe the performance of a range of diagnostic and typing tools for FMDV on a panel of vesicular samples collected in northern Tanzania (East Africa, EA) during 2012–2018. Specifically, we tested these samples with a real-time RT-PCR targeting 3D sequence for pan-FMDV detection; an FMDV monoclonal antibody-based antigen (Ag) detection and serotyping ELISA kit; virus isolation (VI) on LFBKαVβ6 cell line; and a panel of four topotype-specific real-time RT-PCRs, specifically tailored for circulating strains in EA. The 3D real-time RT-PCR showed the highest diagnostic sensitivity, but it lacked typing capacity. Ag-ELISA detected and typed FMDV in 71% of sample homogenates, while VI combined with Ag-ELISA for typing showed an efficiency of 82%. The panel of topotype-specific real-time RT-PCRs identified and typed FMDV in 93% of samples. However, the SAT1 real-time RT-PCR had the highest (20%) failure rate. Briefly, topotype-specific real-time RT-PCRs had the highest serotyping capacity for EA FMDVs, although four assays were required, while the Ag-ELISA, which was less sensitive, was the most user-friendly, hence suitable for any laboratory level. In conclusion, when the four compared tests were used in combination, both the diagnostic and serotyping performances approached 100%.

Immunoglobulins are the primary protective products in human milk and are responsible for transferring maternal pathogen memory to the infant, providing protection by binding to recognized pathogens and inhibiting virulence. To better understand potentially protective/anti-infective compounds in human milk, the establishment of human milk–tailored analytical approaches is crucial, as most contemporary analytical methods have been optimized for plasma or serum. One of the most prominent immunoglobulins in human milk is secretory immunoglobulin A (sIgA), which may be relevant for the protection of breastfed infants from harmful pathogens. Advanced sIgA detection methods can help monitor the immune status and development of the mother-infant dyad. We therefore developed an enzyme-linked immunosorbent assay (ELISA) sIgA method for the quantitative analysis of IgA plus secretory component (SC), validated with sIgA standards and substantiated by mass spectrometry (MS)–based proteomics. A very strong correlation was observed between the MS-detected IgA1 and the human milk–specific sIgA ELISA (r = 0.82). Overall, the MS data indicate that the developed human milk sIgA ELISA does not differentiate between sIgA1 and sIgA2 and is, therefore, a reflection of total sIgA. Furthermore, our MS data and the human milk–derived sIgA ELISA data are better correlated than data derived from a standard serum IgA ELISA kit (relative to MS IgA1 r = 0.82 and r = 0.42, respectively). We therefore propose our human milk–specific sIgA ELISA as an ideal quantitative indicator of total sIgA with advantages over current serum IgA ELISA kits. Supplementary Information The online version contains supplementary material available at 10.1007/s00216-021-03468-4.

Aim of the work: The aim of this study was to evaluate Vimentin, Fetuin-A, and lipoprotein(a) level in the sera of Acute myocardial Infarction (AMI) patients and to determine its relation with disease activity and severity. Patients and methods: Fifty one adult AMI patients (26 females and 27 males) and 41 healthy subjects (23 males and 18 females) matching age serving as the control group, were included in this study. Five milliliters of blood were collected from patients and controls, then centrifuged and the serum was separated and frozen as aliquots at -20 co until use. Measurement of the serum Vimentin, Fetuin-A, and lipoprotein(a) level in AMI patients and controls was done using the ELISA Kit. Results: Serum level of vimentin, and Lp(a)were significantly increased in AMI patient as compared to control group. Serum fetuin-A, albumin, and TP level in RA patient was significantly lower than control group. Hence present study concluded that vimentin, fetuin-A, and Lp(a) could be a useful marker for disease activity in AMI.

The reported incidence of pertussis in European countries varies considerably. We aimed to study specific Bordetella pertussis seroprevalence in Europe by measuring serum IgG antibody levels to pertussis toxin (anti-PT IgG). Fourteen national laboratories participated in this study including Belgium, Denmark, Finland, Greece, Hungary, Italy, Lithuania, Malta, Norway, Poland, Portugal, Romania, Spain, and Sweden. Each country collected approximately 250 samples (N = 7903) from the age groups 20-29 years (N = 3976) and 30-39 years (N = 3927) during 2010-2013. Samples were anonymous residual sera from diagnostic laboratories and were analyzed at the national laboratories by a Swedish reference method, a commercial ELISA kit, or were sent to Sweden for analysis. The median anti-PT IgG concentrations ranged from 4 to 13.6 IU/mL. The proportion of samples with anti-PT IgG ≥100 IU/mL, indicating a recent infection ranged from 0.2% (Hungary) to 5.7% (Portugal). The highest proportion of sera with anti-PT IgG levels between 50 and

Background Toxocariasis is a zoonotic disease in human with worldwide distribution. Recently a relationship between toxocariasis and some autoimmune diseases has been considered. We aimed to investigate the seroprevalence of toxocariasis among multiple sclerosis (MS) and rheumatoid arthritis (RA) patients in comparison with healthy persons. Materials and methods In the present study, 96 blood samples from MS patients, 50 blood samples from RA patients and 100 blood samples from healthy persons were collected. Collected samples were examined to detect anti-Toxocara antibodies using commercial ELISA kit. Results The most of the participants in present study were females (69.1%). The average age of MS patient, RA patients and healthy persons were 37.5, 47.6 and 42.1 years old, respectively. Anti-Toxocara antibody was detected in serum of 8 out of 96 (8.3%) MS patients and 3 out of 100 (3%) healthy persons while anti-Toxocara antibodies were not detected in serum of RA cases. Conclusions This study indicates that there was no significant correlation between IgG antibodies against Toxocara and variables such as gender and age. Based on the statistical analysis, there was no significant difference in Toxocara seropositivity between our control population with MS and RA patients.

This cross-sectional study aimed to study animal, farm, and within-farm seroprevalence of C. burnetii and to identify associated risk factors in goat and sheep farm in northern Jordan. Questionnaire was developed to collect information about risk factors and farms management practices. Blood samples from 730, ≥ 1-year-old females (goat n = 250; sheep n = 480) were randomly collected from 20 goat herds and 40 sheep flocks. IDEXX ELISA Kit was used to detect C. burnetii antibodies. The overall goat and sheep seroprevalence level was 32.5% (237/730) and was significantly higher in goats (43.3%, 108/250; 95% CI 37-49.6) than sheep (27%, 129/480; 95% CI 29.1-36.2) (χ2 test, p ≤ 0.001). Eighty percent (16/20) of goat herds and 60% (24/40) of sheep flocks had at least one seropositive animal (p ≥ 0.05). The average within goat herds and sheep flock seroprevalence were 36.4% (ranged: 0-91%) and 23.4% (ranged: 0-82%), respectively. Multivariate logistic regression model revealed that seroprevalence increased 1.79 times in goat herds compared with sheep flocks, 3.2 times more in farms containing ≥ 100 animals, and 1.7 times higher in farms with their animals that were ≥ 2 years of age than in farms with their animals that are < 2 years of age. In addition, seroprevalence significantly increased 1.52 times in farms loaning bucks or rams during breeding season and 1.63 times in farms containing cats on premises (p ≤ 0.05). Farm biosecurity measures are essential to prevent introduction and minimize transmission of C. burnetii infection to humans and animals.

Pregnancy is a special condition where a female lodges one or more young ones within her uterus. It is maintained by various endocrine physiology and metabolic changes between maternal and fetal circumstances. Space dine secreted by corpus luteum increases extraordinarily throughout the pregnancy, estrogen increases rapidly during first trimester and prolonged lifespan of corpus luteum and small quantity of estrogen prevents prostaglandins pulsatic secretion. We reviewed available literature to evaluate the accuracy of progesterone ELISA test kit in pregnancy diagnosis in farm animals. We found varying accuracy in global and Nepalese context but found that this is an important tool for early pregnancy diagnosis and infertility monitoringwith high accuracy which contributes to increase economic efficiency of a farm. This kit has been used for qualitative and quantitative study of progesterone to understand the reproductive status of animals. Therefore, its use is increasing in globally and in Nepal. This kit has been used at various breeding centers, and livestock service centers of Nepal. However, wider level use is still difficult due to lack of infrastructuresand subsistence farming.

We present the comparative study of plasma chromogranin A (CgA) levels measured in 232 patients with different types of neuroendocrine tumors (NETs) and 66 healthy individuals using enzyme immunoassay Chromogranin A ELISA kit (Dako). CgA levels were significantly higher in patients with all types of NETs than in healthy subjects. CgA secretion were highly variable. The study demonstrate high diagnostic sensitivity of CgA, which was 80.6 % in overall group of NETs with specificity 98.5 %. We confirmed the prognostic significance of CgA, in this case the high basal levels of CgA (above 100 U/l) were significantly associated with less favorable prognosis of progression free survival after different types of treatment. The data confirms high efficiency of CgA as biomarker, which measurement enhances the accuracy of diagnosis and prognosis of NETs.

The selective properties of a solution of oligonucleotide specific to IL-6 on the concentration of IL-6 in mixed saliva of patients with oral inflammatory processes were studied using SDS-PAGE by electrophoresis and enzyme immunoassay. The application of these methods showed that in the mixed saliva of patients after rinsing with a solution of an oligonucleotide specific for IL-6, the amount of IL-6 decreases. The ELISA Kit and 20% SDS-PAGE showed the highest sensitivity to determine the concentration of IL-6 in saliva, which should be considered in clinical laboratory practice.

Background and Objectives: Alveolar and cystic echinococcosis (AE and CE) are caused by the larval stages of Echinococcus multilocularis and Echinococcus granulosus, respectively. This study aimed to determine the prevalence of human AE and CE diseases among the tribes, livestock breeders, and farmers in Moghan plain, northwest of Iran. Materials and Methods: Screening of hydatid cystic fluid antigen (HCF-Ag) was done by enzyme-linked immunosorbent (ELISA) kit. Briefly, HCF-Ag was collected, and the prevalence of the disease was evaluated using specific antigens for AE and CE (Ag-5, Ag-B, and Em2+) and questionnaires. A total of 2453 serum samples were randomly collected from normal populations of five different areas of Moghan plain. The serology was evaluated using ELISA kit and specific antigens for AE and CE. Results: Of the 2453 serum samples, 36 were positive for AE (1.46%). The infection rate was higher in males than females (1.94% vs. 0.94%). The age group of 4–19 years had the lowest and 40–59 years had the highest infection rate (0.2% vs. 2.5%). Among CE-positive serum samples, 178 samples were positive to Ag-5 (7.3%) and 167 samples were positive to Ag-B (6.8%). Conclusion: Our findings showed that the highest infection rates were in the Borran and Eivazlou regions, and the lowest level of infection was for Parsabad area of Moghan plain. In areas with higher contamination, the risk factors associated with CE and AE were the water quality, the method of washing vegetables, and occupational and hydatid disease knowledge of the inhabitants.

Toxoplasmosis, caused by T. gondii, is an important zoonosis worldwide. In Ghana, information on the disease in humans abounds but scanty in animals. This study was therefore conducted to estimate the seroprevalence of T. gondii infection sheep and goats sampled from the Kumasi Abattoir in Ashanti Region, Ghana. A total of 347 serum samples collected from 170 sheep and 177 goats were analyzed for the presence of T. gondii antibodies using a commercial ELISA kit. Results of this study estimated the seroprevalence of 23.7% in goats an, 35.9% in sheep. In sheep, 24 (35.82%) out of a total of 67 male samples were positive and 37 (35.92%) out of a total of 104 female samples were positive while in goats, 6 (8.2%) bucks out of a total of 73 were positive while 36 (34.6%) does out of a total of 104 were positive. There was a significant difference in the rate of seropositivity of female goats (p-value 0.01). This study confirms the existence of T. gondii infection in small ruminants in Ghana and it showed that sheep and dogs are more at risk to T. gondii infection hence meat from such animals could be a potential risk to public health if consumed raw or undercooked.

Serum specimens obtained from 680 individuals were examined to measure the amounts of pepsinogens 1 and 2, anti-CagA antibodies, and anti-Helicobacter pylori antibodies. We examined sera obtained from 610 Mongolian individuals living in the capital city, Ulaanbaatar. Seventy serum specimens were collected from Japanese people who were health-screened: These were stored at the gastroenterology laboratory of Jichi Medical University. The sera of the Japanese people were used as a control specimen. Two enzyme-linked immunosorbent assay (ELISA) kits, an E-plate ELISA kit from Eiken Chemical Co., Ltd. (Tokyo, Japan), and a Biohit ELISA kit from Biohit Oyj (Helsinki, Finland), were used for the detection of anti-H. pylori IgG antibodies in the sera of the 610 Mongolian people. An ELISA kit EIA-4138 from DRG Instruments GmbH (Germany) was used for the detection of anti-CagA IgG antibodies in the serum specimens. Serum pepsinogens were detected by an ELISA kit from Biohit Oyj. Of the 610 serum specimens, 385 specimens tested positive for the detection of anti-H. pylori antibodies using the two ELISA kits, and 47 tested negative. For the detection of anti-H. pylori antibodies by the Biohit ELISA kit, 560 and 50 specimens were positive and negative, respectively. The ratio of serum pepsinogen 1/2 was statistically lower (p &lt, 0.0001) in the H. pylori-positive (560 specimens) than in the H. pylori-negative (50 specimens) specimens. However, the levels of serum pepsinogen 1 had no statistical significance (p = 0.465) between the specimens of the H. pylori-positive and -negative specimens. The ratio of serum pepsinogen 1/2 was 6.74 &plusmn, 0.12 in the H. pylori-positive specimens, whereas the ratio of serum pepsinogen 1/2 was 12.69 &plusmn, 1.02 in the H. pylori-negative specimens. This study demonstrated the high prevalence of H. pylori infection in Mongolian people, including young generations, and the people infected with H. pylori possessed low pepsinogen 1/2 ratios, indicating atrophic gastritis. The serological examinations by the two ELISA kits did not consistently reflect the prevalence of H. pylori infection in Mongolian people.

Aims/Introduction It is reported that interfering substances in the blood might influence the value for measurement of active glucagon‐like peptide‐1 (GLP‐1) in human plasma. Solid phase extraction (SPE) pretreatment is recommended to reduce their influence, but it requires a lot of cost and time. However, there is little investigation about causative inhibitory substances and about methods that can replace solid phase extraction. In the present study, we aimed to seek the candidate of the substances that might interfere with an active GLP‐1 enzyme‐linked immunosorbent assay (ELISA). Materials and Methods Two kinds of active GLP‐1 ELISA kits using different antibodies, plural extraction carriers and elution solutions were used to evaluate the SPE method. Active GLP‐1 concentration was compared with or without SPE, and with or without a heterophilic blocking tube. Results Active GLP‐1 values were often higher without SPE compared with those with SPE pretreatment. This difference was eliminated by pretreatment with a heterophilic blocking tube or ELISA kits that did not use a mouse monoclonal antibody, and was independent of SPE. Conclusions Substances absorbed to a heterophilic blocking tube carrier might interfere with an active GLP‐1 immunoassay. Solid‐phase extraction treatment is required for measurement of active GLP‐1 by an ELISA kit affected by heterophilic antibodies.

The goal of research was development of competitive ELISA kit for detection of genetically modified plants resistant to glyphosate. As the source for gene isolation we have used leaves of GM soybean RoundUp Ready, line 40-3-2 (Monsanto). Using polymerase chain reaction the gene encoding the enzyme 5-enolpiruvilshykimat-3-phosphate synthase (CP4 EPSPS) was isolated, which provides resistance to the herbicide glyphosate. Isolated gene has been cloned into plasmid vector pET24(a), creating genetic construct pET-CP4EPSPS. E.coli (BL 21(DE3)) has been transformed by genetic construct and over expressed after induction by 0.5 mM IPTG. Recombinant enzyme CP4 EPSPS with molecular weight 45 kD was purified as inclusion bodies and used as antigen for 96-well polystyrene plates immobilization. Monoclonal antibodies CP4 EPSPS mAb3 (Artron BioResearch Inc., Canada) was used for synthesis of horseradish peroxidase conjugate and receiving diagnostic reagent. We have shown that in the competitive ELISA we can detect at least 0.1% of GM plants resistant to glyphosate, on the example of soybean (line RR 40-3-2), moreover we can also detect rapeseed (line GT73) and sugar beet (line H7-1).

Celiac disease and nonceliac gluten sensitivity are provoked by the consumption of gluten from wheat, barley, rye, and related grains. Affected individuals are advised to adhere to gluten-free diets. Recently, gluten-free foods have become a marketing trend with gluten-free options both in packaged foods and in restaurants and food service establishments. Pasta is one of the primary gluten-containing foods in diets in North America and Europe. Gluten-free pasta formulations are commercially available. In restaurants, multiple pasta dishes are often prepared simultaneously in large pots with multiple compartments and shared cooking water. The objective of this study was to determine whether gluten transfer occurs between traditional and gluten-free pasta when cooked simultaneously in the same water. Pasta was boiled in a commercial, four-compartment, 20-qt (18.9-L) cooking pot containing three batches of traditional penne pasta and one batch of gluten-free penne pasta. The amount of pasta (dry weight) was either 52 g (recommended serving size) or 140 g (typical restaurant portion). Five consecutive batches of pasta were boiled, and cooking water and gluten-free pasta were sampled at completion of cooking. Water and gluten-free pasta samples were tested for gluten with the Neogen Veratox for Gliadin enzyme-linked immunosorbent assay kit. Gluten concentrations were low (20 ppm) in both water and gluten-free pasta samples through five 52-g batches. Gluten concentrations in the 52-g gluten-free pasta samples slowly increased through five batches but were never20 ppm. During cooking of the 140-g gluten-free pasta samples, the gluten concentrations in the cooking water increased with each batch to50 and80 ppm after the fourth and fifth batches, respectively. The gluten concentrations in the 140-g gluten-free pasta samples approached 20 ppm by the fourth batch and reached nearly 40 ppm after the fifth batch. Although gluten transfer does not occur at a high rate, gluten-free pasta should be prepared in a separate cooking vessel in restaurant and food service establishments.

Daud D, Dewa MSAM, Mahbob ENM, Razak WRWA. 2021. Short Communication: Phytochemical diversity and bioactivity of Malaysian Premna cordifolia (Lamiaceae). Biodiversitas 22: 3245-3248. The aims of the present study are to determine the chelation potential and protective effect of Premna cordifolia leaves ethanolic extract in lead intoxicated mice. Twenty-four male mice were divided into four groups with six animals each. Group 1 was healthy mice and received 2 ml/kg bwt distilled water. Group 2 was healthy mice and supplemented with 200 mg/kg bwt P. cordifolia leaves ethanolic extract. Group 3 was lead intoxicated mice and received 2 ml/kg bwt distilled water. Group 4 was lead intoxicated mice and supplemented with 200 mg/kg bwt P. cordifolia leaves ethanolic extract. All treatments were given for 14 days. Chelation activity was determined by measuring the concentrations of lead in the liver, kidney and testis by using atomic absorption spectroscopy. Sperm quality was tested according to World Health Organization manual and testosterone was estimated by using ELISA kit. Spectroscopic data showed that P. cordifolia leaves ethanolic extract was able to lower lead concentrations in liver, kidney and testis of lead intoxicated mice. Following a decrease in lead concentrations, sperm quality and testosterone levels were restored in lead intoxicated mice supplemented with P. cordifolia leaves ethanolic extract. In conclusion, P. cordifolia leaves ethanolic extract has the potential to be developed as an alternative remedy for treating lead poisoning effects.

This study was conducted to investigate the reproductive pattern of bitches by estimation the optimumtime of ovulation and kisspeptin level. The study included 245 bitches (different breeds), from March/2019to the end of February/2020.Canine ovulation timing kits was applied in order to determine the ovulationtime. For estimation of Kisspeptin, blood collection of 80 samples allocated approximately 7 samples foreach monthby using ELISA kit. The resultsrevealed the time of ovulation ranged from 2 to 5 days after thebeginning of the estrus phase.The results showed that concentration of kisspeptin during different phases ofthe estrous cycle was (257.01±11.08, 262.93±9.15, 227.98±14.91 and 244.89±11.99) pg/mL for Proestrus,Estrus, Diestrus, and Anestrus respectively with no significant (p>0.05) differences among different estrouscycle phase. The concentration of kisspeptin in which distributed according to the season showed that itwas (218.26±11.92, 254.12±10.09, 246.00±10.22, and 287.11±9.00) pg/ml for Spring, Summer, Autumn,and Winter, respectively and there was a significant difference (p

Goat Warble Fly Infestation (GWFI) is also known as subcutaneous myiasis caused by Przhevalskiana silenus (Diptera: Oestridae). It is widely distributed in tropical and sub-tropical areas of the world. In goats, WFI is usually detected through conventional procedure which underestimated the infestation. The current study was designed to determine the serodiagonsis of GWFI (through IDEXX Hypodermosis serum antibody test) and also aimed to investigate its seroepizootiological profile in Pothwar region, Pakistan from 2013-14. The results showed that average seropositivity (ELISA kit) of GWFI was 18.5% whereas, it was 11% by using conventional procedure (Palpation method) depicting a significant difference (p0.05). Higher seropositivity (30.8%) was observed in Jhelum district as compared to e Attock district (6%). The L1 larvae were found in September, while nodules start appearing in October to December and last until the end of February. The month wise peaks of optical density (OD) was higher in December which gradually decrease along with the end of winter season. The prevalence of GWFI revealed no significant difference among three host breeds (Jattal, Beetal and Tedy). According to the results, high infestation rate (28%) was observed in young animals of age group1 year as compared to old animals (2 years). Topographically, hilly areas (33%) provide favourable climatic conditions for the propagating of larval stages. Sex difference showed no significant difference. The seroprevalence varied significantly with respect to age, month, districts and topographical location. The current study proved that serologic diagnosis (commercial ELISA kit) as more sensitive and accurate for timely diagnosis of GWFI than traditional method. The information on the epizootiology of P. silenus in goats of Pothwar region would help in devising effective control strategies.

The present study describes the development of a truncated recombinant peste des petits ruminants virus (PPRV) nucleoprotein (rPPRV-NPN) and its polyclonal antibodies-based immuno-diagnostic assay, Avidin-Biotin (AB) recombinant nucleoprotein competitive ELISA (ABrC-ELISA) for the detection of PPRV antibodies in the sheep and goats. The PPRV N-terminal immunogenic region (1-266 aa) of nucleoprotein (NPN) coding sequence was amplified and cloned into the pETite vector. The rPPRV-NPN with a molecular weight of ∼ 30 kDa was expressed in E. coli, purified, and characterized by SDS-PAGE and immunoblot using standard PPRV specific sera. The Ni-NTA affinity-purified rPPRV-NPN as coating antigen and its hyperimmune serum as competitive antibodies raised in guinea pigs were evaluated as diagnostic reagents in ABrC-ELISA using the known standard panel of sera. The threshold (cut-off) Percentage Inhibition (PI) value was determined as 45 (mean ± 3 SD) based on the reactivity of the known sheep and goats sera to PPRV antibodies [negative (n = 140) and positive (n = 98)] and the assay had a sensitivity of 97 % (95 % Confidence Interval (CI): 91.3-99.4 %) and specificity of 100 % (95 % CI: 97.4-100 %) with an excellent Area under curve (AUC) of 0.997 (95 % CI: 0.99-1.0). On evaluation of diagnostic performance of the assay using the sheep and goats sera (n = 391) from vaccinated, infected, and non-vaccinated animals, the ABrC-ELISA showed the relative diagnostic sensitivity of 95.88 % (95 % CI: 92.56-98.01 %) & 98.77 % (95 % CI: 96.43-99.74 %) and diagnostic specificity of 97.97 % (95 % CI: 94.19-99.58 %) & 90.54 % (95 % CI: 84.64-94.73 %) against indigenous PPR competitive ELISA kit & IDvet Screen® PPR Competition kit, respectively. The study showed that ABrC-ELISA is rapid, sensitive, and specific and can be a better alternative assay for the detection of the PPRV antibodies in the sera of small ruminants for serosurveillance / seromonitoring of PPR not only at the eradication and post-eradication phases in the disease-controlled endemic countries but also in the PPR non-endemic countries.

Se determinó la relación entre los factores socioeconómicos-epidemiológicos y la seroprevalencia de toxoplasmosis en gestantes atendidas en los Hospitales “Felipe Arriola” y “César Garayar” de la ciudad de Iquitos, Perú en el 2009. El estudio fue descriptivo y transversal en 355 gestantes de 15 a 45 años de edad que asistieron a su control pre-natal, seleccionadas intencionalmente. Se aplicó una encuesta} socioeconómica-epidemiológica sobre datos personales, económicos, sanidad de la vivienda, cohabitación con gatos y otros animales, y hábitos alimenticios. Se determinaron la IgG e IgM anti Toxoplasma gondii utilizando un kit comercial de ELISA. Para el análisis estadístico se utilizó la prueba X2 de Mantel-Heanszel. La seroprevalencia general de toxoplasmosis fue de 97,6% en las gestantes del Hospital “Felipe Arriola” y 97,4 % en las del Hospital “Cesar Garayar”. La frecuencia de anticuerpos IgG fue 94,5% y 86,8% en gestantes de los hospitales “Felipe Arriola” y “César Garayar”, respectivamente. Se registraron bajas frecuencias de anticuerpos IgG e IgM, 3,1% en el Hospital “Felipe Arriola” y 10,5% en Hospital “César Garayar”. Los factores socioeconómicosepidemiológicos relacionados significativamente con la seroprevalencia de toxoplasmosis fueron: ingesta de frutas y verduras sin lavar (p=0,002) en gestantes del Hospital “Felipe Arriola”, y vivir en vivienda rústica (p=0,010) y cohabitar con otros animales (p=0,02) en gestantes del Hospital “César Garayar”. Se concluye que la seroprevalencia de toxoplasmosis fue alta en las gestantes de ambos hospitales de Iquitos, Perú el año 2009.

An instance of hormone assay method flaw is reported. In this journal Chronobiology International, two papers appeared in which an ELISA method for human serum or plasma was utilized for blood serum of horse and sheep, respectively. From our testing, it is resulted that such method does not work at all for equine, sheep and other animal species. The use of commercial hormone assay kits for heterologous species always needs a careful validation procedure. First, the same hormone molecule by different species could not share enough homology to be regognized by and react with antibodies utilized in the method. Furthermore, even with a full overlap of the molecules, possible interferences by other components of the sample (matrix effect) have to be considered.

Plasma IL-17A detection in Langerhans Cell Histiocytosis (LCH) is currently a source of debate. Indeed, 500-P07G (PeproTech) and 41802 (R&D Systems) anti-IL-17A antibodies have been suspected to recognize nonspecific proteins. To resolve this discrepancy, we set up two new ELISAs by using 41802 or neutralizing eBio64CAP17 (eBioscience) capture monoclonal antibodies that we compared to the commercial PeproTech ELISA kit. The three ELISAs, called E_500-P07G, E_41802 and E_eBio64CAP17, differ in their anti-IL-17A capture antibodies: either polyclonal, monoclonal or neutralizing monoclonal antibodies, respectively. Here, we show that these ELISAs had a similar capacity to specifically detect recombinant or native human IL-17A. However, a significantly lower plasma IL-17A detection was obtained with E_41802 compared to the two other ELISAs. Both E_500-P07G and E_eBio64CAP17 showed similar results. Consequently, we propose that the use of E_500-P07G and E_eBio64CAP17 may ensure more accurate and reliable results in the context of LCH studies. The highest plasma IL-17A levels in LCH patients compared to controls detected by both E_500-P07G and E_eBio64CAP17 ELISAs led us to propose these latter as reference techniques to investigate IL-17A as a potential new biomarker in LCH.•The customization of a new E_eBio64CAP17 ELISA is suitable to detect human IL-17A.•E_eBio64CAP17 ELISA protocol differs only in the anti-IL-17A capture antibody compared to the commercial E_500-P07G PeproTech kit.•Data generated using the E_eBio64CAP17 ELISA are consistent with the PeproTech kit., Graphical abstract Image, graphical abstract

T-2 is a common mycotoxin contaminating cereal crops. Chronic consumption of food contaminated with T-2 toxin can lead to death, so simple and accurate detection methods in food and feed are necessary. In this paper, we establish a highly sensitive and accurate method for detecting T-2 toxin using AlphaLISA. The system consists of acceptor beads labeled with T-2-bovine serum albumin (BSA), streptavidin-labeled donor beads and biotinylated T-2 antibodies. T-2 in the sample matrix competes with T-2-BSA for antibodies. Adding biotinylated antibodies to the test well followed by T-2 and T-2-BSA acceptor beads yielded a detection range of 0.03–500 ng/mL. The half-maximal inhibitory concentration was 2.28 ng/mL and the coefficient of variation was

Among &ldquo, extra-criteria&rdquo, antiphospholipid antibodies (aPL), anti-phosphatidylserine/prothrombin (aPS/PT) antibodies, are considered a part of risk assessment strategies when investigating patients suspected of having antiphospholipid syndrome (APS). aPL detection is currently performed by solid-phase assays to identify anti-cardiolipin (aCL), anti-&beta, 2glycoprotein I (a&beta, 2GPI) and aPS/PT antibodies, but new techniques are emerging. Among these, particle-based multi-analyte technology (PMAT), which allows the full automation and simultaneous digital detection of autoantibodies and proteins, including IgG, IgA and IgM isotypes of aCL, a&beta, 2GPI and aPS/PT. The aim of this study was to investigate the agreement of aPS/PT testing between enzyme-linked immunosorbent assay (ELISA) and the PMAT platform. A total of 94 patients were enrolled in the study, including 71 patients with confirmed APS and 23 &ldquo, aPL carriers&rdquo, aPS/PT IgG showed a moderate binomial agreement between ELISA and PMAT (k = 0.57, 95% CI 0.45&ndash, 0.75), and aPS/PT IgM showed a moderate agreement (k = 0.60, 95% CI 0.45&ndash, 0.75). Moreover, when considering the continuous agreement, both aPS/PT IgG and IgM showed a statistically significant correlation between ELISA and PMAT (Spearman&rsquo, s correlation = 0.69, p &lt, 0.001 and 0.72, p &lt, 0.001, respectively). This study demonstrates that PMAT technology is a reliable method for aPS/PT IgG and IgM testing when compared to the available commercial ELISA kit.

Lumpy skin disease (LSD) is an endemic infectious viral disease of cattle in African and Middle East countries. A cross-sectional study was conducted to determine the seroprevalence of LSD among cattle in Northern Egypt and assessed the associated risk factors with LSD infection. A total of 1000 sera samples were collected from cattle and examined serologically with commercial ELISA kit. Overall, the true seroprevalence of LSD among cattle in Egypt was 19.5% (n = 180). The true seroprevalence rate was varied significantly between different geographical studied areas, and the highest level was observed in Kafr El-Sheikh (26.7%) and Gharbia (23.7%) governorates. Moreover, the risk of getting LSD infection was increased among Holstein breed (OR = 4.586; 95%CI, 1.83–11.48) and adult cattle (OR = 2.498; 95%CI, 1.17–5.32) during summer season (OR = 7.303, 95%CI: 3.97–13.42). Furthermore, communal grazing (OR = 1.546; 95%CI, 0.91–2.60), communal water points (OR = 3.283; 95%CI, 2.11–5.09), introduction of new animal (OR = 2.216; 95%CI, 1.32–3.71), and contact with other animals (OR = 3.401; 95%CI, 1.62–7.10) were identified as significant risk factors for the occurrence of LSDV infection in cattle. Moreover, the present study revealed that no significant (P > 0.05) association between sex or type of herd and occurrence of LSD infection.

Detection of IgA antibody against Mycobacterium avium complex (MAC) glycopeptidolipid (GPL) has recently been shown to improve the diagnosis of MAC pulmonary disease but has yet to be tested in disseminated Non-tuberculous mycobacteria (NTM) infection. In this study, we address the diagnostic efficacies of an anti-GPL-core ELISA kit in disseminated lymphadenopathy patients positive for NTM culture and anti-IFN-γ autoantibodies. The study was conducted in a tertiary referral center in northeastern Thailand and patients with NTM, tuberculosis, melioidosis, and control subjects were enrolled. Plasma immunoglobulin A (IgA) and G (IgG) antibodies against GPL-core were detected in the subjects and the specificity and sensitivity of the assay was assessed. Anti-GPL-core IgA and IgG levels were significantly higher in NTM patients than other groups (p < 0.0001). Diagnostic efficacy for NTM patients using anti-GPL-core IgA cut-off value of 0.352 U/ml showed good sensitivity (91.18%) and intermediate specificity (70.15%). Using a cut-off value of 4.140 AU/ml for anti-GPL-core IgG showed the same sensitivity (91.18%) with increased specificity (89.55%) and an 81.58% positive predictive value. Most patients with moderate levels (4.140–7.955 AU/ml) of anti-GPL-core IgG had rapidly growing mycobacteria (RGM) infection. Taken together, the detection of anti-GPL-core antibodies could provide a novel option for the diagnosis and management of disseminated NTM infected patients.

Identifying Hepatitis E virus (HEV)-positive pig farms is important to implement surveillance programs for this emerging zoonotic agent. The aim of this study was to evaluate the use of serosanguineous fluids obtained as part of castration practice (processing fluids (PFs)) to detect anti-HEV antibodies in newborn piglets. Ninety-five paired serum and PF samples were collected from piglets of 29 different litters and tested with a commercial ELISA kit. A significant positive correlation (Spearman’s rho: 0.600, p &lt, 0.01) was found between anti-HEV antibodies in serum and PF samples. In 26 out of 29 litters (89.7%), there was at least one positive piglet in the serum. Sixteen litters out of 29 (55.2%) were also positive in PFs. To simulate the use of PF as pooled samples, the limit of detection of the ELISA was assessed mixing the PF sample with strong, medium, medium-weak and weak ELISA titres with 3, 4, 5 and 6 negative PF samples. Our results suggest that it is still possible to identify a positive PF pool when at least one individual PF sample with medium or strong antibody levels is mixed with 5 or 6 individual negative PF samples. The detection of anti-HEV maternal-derived antibodies in PF confirms a past exposure of sows to the virus. PF may represent a rapid, noninvasive and economical tool to identify HEV-positive farms.

Dengue virus (DENV) and Zika virus (ZIKV) belong to the flavivirus genus and are antigenically closely related. They also share the same mosquito vector and can cause similar symptoms upon infection. However, DENV and ZIKV infections lead to different clinical sequelae and treatments, therefore, clinicians need rapid and accurate diagnostics capable of distinguishing between the two diseases. Methods: We employed the immuno-magnetic assay technology on a microfluidic cartridge (ViroTrack Sero Zika IgG/IgM) for diagnosis of ZIKV infection based on the aggregation of magnetic nanoparticles. We carried out three serological studies including samples from the Dominican Republic, USA, and Nicaragua, aimed at detecting ZIKV-specific IgG and IgM using the ViroTrack Sero Zika IgG/IgM test. Results: The seroconversion results were comparable with ZIKV IgG and IgM reactivity measured by the commercial ZIKV ELISA kit. The sensitivity and specificity for both ZIKV IgG and IgM tested by the ViroTrack Sero Zika IgG/IgM was approximately 98% and 93%, respectively. Conclusion: Serological detection of ZIKV infection by the new ViroTrack Sero Zika IgG/IgM test shows promising performance and limited cross-reactivity with DENV.

Background Detection of Clostridium perfringens enterotoxin (CPE) is critical for disease surveillance; however, commercial testing kits produce contrasting results. Methods We examined the cause of the differing results from a reversed passive latex agglutination (RPLA) assay (PET-RPLA Toxin Detection Kit) and an enzyme-linked immunosorbent assay (C. perfringens Enterotoxin ELISA Kit) using 73 human norovirus-positive fecal samples from gastroenteritis patients across 22 episodes in Japan. Results CPE was detected in 39/73 samples using the RPLA method; however, ELISA-based examination of 10 RPLA-positive samples produced negative results. Moreover, cpe was not detected in any of the RPLA-positive (n = 32) or -negative (n = 5) samples, and C. perfringens was only isolated from one RPLA-positive sample. Conclusions An ELISA-based testing approach may be more reliable than RPLA assays for CPE detection from human fecal samples. These findings may also be applicable to the detection of other foodborne diseases.