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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%.

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.

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.

People with some occupational or recreational activities, such as hunters and veterinarians, may have increased risk to be infected by the hepatitis E virus (HEV). The aim of the present study was to establish whether forestry workers could be considered at a higher risk of HEV infection than a control group. One hundred and fifty sera from forestry workers and a control group of 85 sera were analysed by anti-HEV IgG antibodies detection using a commercial ELISA kit. The anti-HEV IgG seroprevalence was 14% for forestry workers and 9.4% for the control group. Comparing the risk of HEV infection in the two groups, there was no difference in the odds ratio. However, the seroprevalence in older subjects was higher in the forestry workers than in the control group. Two sera from forestry workers were also positive for anti-HEV IgM, and, in one of them, HEV-RNA was detected. Our findings showed an increase of seroprevalence with age, which is likely to reflect cumulative exposure to HEV over time. The occupation of forestry workers did not seem to be associated with a higher risk of HEV infection. The study provided new insights into the risk of acquiring HEV in occupational exposure workers with open-air activities.

Lyme disease (LD), caused by infection with Borrelia burgdorferi, is the most common tick-borne infection in many regions of Eurasia. Antibody detection is the most frequently used laboratory test, favoring a two-step serodiagnostic algorithm, immunoenzymatic detection of antibodies to C6 has been shown to perform similarly to a standard two-step workflow. The aim of this study was the performance evaluation of the C6 Lyme ELISA kit compared to a standard two-step algorithm in three laboratories located in the northeastern region of Italy which cater to areas with different LD epidemiology. A total of 804 samples were tested, of which 695 gave concordant results between C6 testing and routine workflow (564 negative, 131 positive). Wherever available, clinical presentation and additional laboratory tests were analyzed to solve discrepancies. The C6 based method showed a good concordance with the standard two-step algorithm (Cohen&rsquo, s &kappa, = 0.619), however, the distribution of discrepancies seems to point towards a slightly lower specificity of C6 testing, which is supported by literature and could impact on patient management. The C6 ELISA, therefore, is not an ideal stand-alone test, however, if integrated into a two-step algorithm, it might play a part in achieving a sensitive, specific laboratory diagnosis of LD.

Background: Exposure to ETS (environmental tobacco smoke) is one of the most toxic environmental exposures. Objective: To investigate the association of ETS with physiological, biochemical, and psychological indicators, as well as with urine antioxidant capacity (AC) and oxidative damage to lipids in a pilot sample of healthy pregnant women. Methods: Exposure to ETS was investigated via a validated questionnaire, and urine cotinine and the marker of oxidative damage to lipids via 8-isoprostane concentrations using an ELISA kit. Urine AC was determined by the spectrophotometric Trolox-equivalent antioxidant capacity (TEAC) method. From a sample of pregnant women (n = 319, average age 30.84 &plusmn, 5.09 years) in 80, the levels of cotinine and oxidative stress markers were analyzed. Results: Among the 80 pregnant women, 5% (7.4% confirmed by cotinine) reported being current smokers and 25% reported passive smoking in the household (18.8% confirmed by cotinine). The Kappa was 0.78 for smokers and 0.22 for ETS-exposed nonsmokers. Pregnant women in the ETS-exposed group had significantly reduced AC compared to both the nonsmoker (ETS&minus, ) and the smoker groups (p &lt, 0.05). Nonsmokers had significantly lower levels of 8-isoprostane than smokers (p &lt, 0.01) and ETS-exposed nonsmokers (p &lt, 0.05). Correlations between urine levels of cotinine and AC were positive in ETS-exposed nonsmokers. Conclusion: A harmful association of active and passive smoking and oxidative stress parameters among pregnant women has been indicated.

A total of 328 agricultural product samples highly suspected to be contaminated, from flour companies, feed companies, and livestock farms throughout China, were surveyed for deoxynivalenol (DON) contamination using a self-assembly enzyme-linked immunosorbent assay (ELISA) kit. An ELISA kit for DON was developed with a 4.9 ng mL&minus, 1 limit of detection (LOD) in working buffer and a 200 ng g&minus, 1 LOD in authentic samples. The DON contamination detection rate was 88.7%, concentrations ranged from 200.9 to 6480.6 ng g&minus, 1, and the highest DON contamination was found in distillers&rsquo, dried grains with solubles with an average of 3204.5 ng g&minus, 1. Wheat bran and wheat were found to be the most commonly contaminated samples, and the corn meal samples had the lowest average DON level. This ELISA kit is a powerful alternative method for the rapid, sensitive, specific, accurate, and high-throughput determination of DON and can meet the maximum requirement levels. This survey suggests that DON contamination in the Chinese market is serious, and the contamination risk deserves attention. Essential preventive measures should be implemented to ensure food safety and human health.

This study was conducted to develop a self-assembled direct competitive enzyme-linked immunosorbent assay (dcELISA) kit for the detection of deoxynivalenol (DON) in food and feed grains. Based on the preparation of anti-DON monoclonal antibodies, we established a standard curve with dcELISA and optimized the detection conditions. The performance of the kit was evaluated by comparison with high-performance liquid chromatography (HPLC). The minimum detection limit of DON with the kit was 0.62 ng/mL, the linear range was from 1.0 to 113.24 ng/mL and the half-maximal inhibition concentration (IC50) was 6.61 ng/mL in the working buffer, there was a limit of detection (LOD) of 62 ng/g, and the detection range was from 100 to 11324 ng/g in authentic agricultural samples. We examined four samples of wheat bran, wheat flour, corn flour and corn for DON recovery. The average recovery was in the range of 77.1% to 107.0%, and the relative standard deviation (RSD) ranged from 4.2% to 11.9%. In addition, the kit has the advantages of high specificity, good stability, a long effective life and negligible sample matrix interference. Finally, wheat samples from farms in the six provinces of Henan, Anhui, Hebei, Shandong, Jiangsu and Gansu in China were analyzed by the kit. A total of 30 samples were randomly checked (five samples in each province), and the results were in good agreement with the standardized HPLC method. These tests showed that the dcELISA kit had good performance and met relevant technical requirements, and it had the characteristics of accuracy, reliability, convenience and high-throughput screening for DON detection. Therefore, the developed kit is suitable for rapid screening of DON in marketed products.

Simple Summary Although cortisol is usually considered the main reference for the assessment of stress, in some animal species it has been shown that prolactin can be used as a biomarker of both acute and chronic stress. Behavioural parameters can also be used to assess the state of welfare and stress. This study was aimed at evaluating the possible relationship between serum prolactin, serum cortisol and behavioural signs of stress in domestic dogs. To reduce the possible influence of some factors, the study was performed on a homogeneous sample formed by 40 castrated male Spanish Greyhound dogs housed in a dog shelter. The weak negative correlation found between serum cortisol and prolactin values agrees with results obtained in other studies, indicating that prolactin response might be an alternative to cortisol response. Abstract Prolactin has been recently regarded as a potential biomarker of both acute and chronic stress in several species. Since only few studies until now have focussed on domestic dogs, this study was aimed at evaluating whether prolactin, cortisol and stress behaviour correlated with each other in sheltered dogs. Both cortisol and prolactin analysis were performed in serum samples through a hormone-specific ELISA kit. For each dog, a stress score was calculated by summing the number of occurrences of stress-related behaviours. The presence/absence of fear during the time spent in the collection room was also scored for each individual. Results revealed a weak negative correlation between cortisol and prolactin levels. Neither of the hormones was correlated with the stress score, nor did their values seem to be influenced by showing fear in the collection room. The weak negative correlation found between cortisol and prolactin values agrees with results obtained in other studies, indicating that prolactin response might be an alternative to cortisol response. This, together with the high serum prolactin levels compared to those reported by other authors for healthy domestic dogs, may indicate that prolactin might be a good biomarker of chronic stress, and although further studies are needed to better understand the potential role of prolactin in the evaluation of canine welfare.

Enzyme-linked immunosorbent assays (ELISA) show extensive application in immunoassays, to detect and monitor protein biomarkers in clinical diagnosis. Nevertheless, the time required and its multiple steps limit its application. We take advantage of a polyethyleneimine (PEI) gold nanoparticle (GNP) coated microwell plate to perform blocking-free ELISA, in which no nonspecific protein adsorption appears on the GNP layer. If the PEI-GNP coated microwell plate and immobilization of captured antibodies on the plate are prepared in advance, such as using an ELISA kit, the whole ELISA process can be finished in less than 2 h. Meanwhile, we have ensured that the GNP layer can preserve the precision and good linearity of ELISA without causing negative effects on the plate.

Bone loss and osteoporosis is a serious health problem worldwide. The impact of osteoporosis is far greater than many other serious health problems, such as breast and prostate cancers. Statistically, one in three women and one in five men over 50 years of age will experience osteoporotic fractures in their life. In this paper, the design and development of a portable IoT-based sensing system for early detection of bone loss have been presented. The CTx-I biomarker was measured in serum samples as a marker of bone resorption. A planar interdigital sensor was used to evaluate the changes in impedance by any variation in the level of CTx-I. Artificial antibodies were used to introduce selectivity to the sensor for CTx-I molecule. Artificial antibodies for CTx-I molecules were created using molecular imprinted polymer (MIP) technique in order to increase the stability of the system and reduce the production cost and complexity of the assay procedure. Real serum samples collected from sheep blood were tested and the result validation was done by using an ELISA kit. The PoC device was able to detect CTx-I concentration as low as 0.09 ng/mL. It exhibited an excellent linear behavior in the range of 0.1–2.5 ng/mL, which covers the normal reference ranges required for bone loss detection. Future possibilities to develop a smart toilet for simultaneous measurement of different bone turnover biomarkers was also discussed.