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Context: Microsporidiosis is an emerging disease in both immunocompetent and immunocompromised patients. Diagnosis of human microsporidiosis is challenging. Parasitological tests using staining methods have limitations for detecting spores in stool samples. The performance of molecular methods for diagnosing human microsporidiosis is satisfactory, but these methods are expensive and time-consuming. Serodiagnosis tests for the detection of antibodies have insufficient specificity. Detecting microsporidia antigens seems to be an appropriate method for diagnosing human microsporidiosis. Therefore, the present study reviewed serological approaches to assess the current status of these methods for diagnosing human microsporidiosis. Evidence Acquisition: All related published articles were searched. The search terms were “microsporidiosis”, “microsporidiasis”, “human microsporidiosis”, and “human microsporidiasis”, combined with the search terms “diagnosis”, “serodiagnosis”, “immunodiagnosis”, “antigen detection”, and “antibody detection”. Data were extracted from the articles that met our eligibility criteria. Immunodiagnosis studies in experimentally infected animals were excluded. Results: Few studies have evaluated the performance of serological tests for diagnosing human microsporidiosis. The IgG response against spore wall protein or polar tube antigens remains at least three years after infection and shows more cross-reactivity with other species. Therefore, the western blot should be conducted as a confirmatory method. Serodiagnosis tests using monoclonal antibodies against microsporidia showed less cross-reactivity and more efficacy compared to polyclonal antibodies. Conclusions: Indirect fluorescent antibody test based on the monoclonal antibody is highly effective in diagnosing microsporidia species in clinical samples.

Microsporidia belong to the intracellular spore-like pathogen, that can cause infection in invertebrates and vertebrates, including humans. Encephalitozoon spp. and Enterocytozoon bieneusi, are important causes of chronic diarrhea, especially in patients with HIV/AIDS. Therefore, in this study, modified trichrome staining (MTS) and nested polymerase chain reaction (nested PCR) methods were used for the diagnosis of common intestinal microsporidia in faecal samples of patients with HIV/AIDS in Zahedan, southeastern Iran, for the first time. Stool samples were collected from 50 HIV/AIDS-infected patients with gastrointestinal symptoms whose infections were confirmed by serology test. Prepared smears from each stool sample were stained using the MTS method. Nested PCR was used to amplify 440 bp and 629 bp fragments of 16S rRNA genes in E. bieneusi and Encephalitozoon spp., respectively. Based on the MTS method and the nested PCR, 8 (16%) and 12 (24%) stool samples were positive, respectively. According to the results of nested PCR, eight, three, and one case were infected with E. bieneusi, Encephalitozoon spp., and both of them, respectively. Findings indicated microsporidiosis in HIV/AIDS-infected patients in Zahedan is an important health problem. Therefore, this opportunistic microorganism in HIV/AIDS-infected patients should be diagnosed using sensitive and accurate methods.

Hydatidosis or cystic echinococcosis (CE) is one of the most common zoonosis diseases. Iran is one of the endemic regions in terms of this disease. For the first time, the present study was conducted to determine the seroprevalence of hydatid cyst in Zahedan rural areas due to the importance of human CE and lack of information in this region. The present study was performed on 551 people referred to seven rural health centers in Zahedan during 2019-2020. Serum samples were collected and analyzed by indirect ELISA method using recombinant antigen B subunit B8/1. Results were analyzed by SPSS (version 22) software and Chi-square test. The CE seroprevalence was 4%. The most positive cases were in the age group of 10-30 years. The highest infection was reported in homemakers. A significant relationship (P-value0.05) was only reported between the seropositivity to hydatid cyst and the presence of dogs in the environment. The present study's findings indicated human hydatid cyst in rural areas of Zahedan is a health problem; moreover, the control and prevention principles and analysis of various epidemiological aspects of this disease should be considered.

Toxoplasmosis is one of the protozoan diseases caused byA total of 1,324 blood samples of patients referred to the health care centers were evaluated using the IgG Toxoplasma ELISA Kit, between October 2019 and August 2021. The obtained data were analyzed through univariable and multivariable regression models.The seropositivity ofThe results of this study indicated significant seropositivity to

Background: Malaria is one of the most serious parasitic diseases and the major health issue in tropical and subtropical countries. Objectives: This study aimed to investigate the type of infection of female Anopheles mosquitoes to sporozoites of Plasmodium species (Plasmodium vivax and Plasmodium falciparum), using nested polymerase chain reaction (PCR) in southeastern Iran. Methods: To determine the infections and Plasmodium species using the nested PCR in Sistan and Baluchestan Province (southeastern Iran), 360 female Anopheles mosquitoes were collected from the control villages of Chabahar and Konarak counties and examined by entomology check stations affiliated with Zahedan University of Medical Sciences in the second half of 2018. The mosquitos were caught manually from indoor places (humans and animals) as well as natural and artificial sites, i.e., pit-shelters. After DNA extraction, the molecular analysis was performed by nested PCR. Results: Of 200 species collected from Daregas, Owraki, and Bahu Kalat in Chabahar, four Anopheles stephensi (2%) were infected with P. vivax, and of 160 samples collected from the rural districts of Konarak, including Zarabad, Heyan, Islamabad, and Joholu, two An. stephensi (1.25%) were infected with P. falciparum. In this study, no mixed infection of Plasmodiumvivax and Plasmodium falciparum were observed. Conclusions: The present study showed that the only species infected with Plasmodium in the aforementioned regions was An. stephensi, which was the most frequent species of Anopheles among other species as well. The nested PCR could be the most suitable method for detecting Plasmodium parasites in malaria vectors, among other diagnostic methods.

Background: In the current study, we described the epidemiological features, clinical presentation, diagnosis and management of patients with suspicion of fascioliasis in Kohgiluyeh and Boyer-Ahmad Province in southwest of Iran. Methods: Overall, 56 patients with suspicion of fascioliasis, based on their clinical signs and symptoms that referred to Clinic of Internal Medicine in Yasuj city, from 2014 to 2016 were enrolled. Demographic data, history of eating aquatic local plants, the chief complains, and laboratory findings were recorded for each patient. Stool samples were obtained from each case for detection of Fasciola eggs. Moreover, blood samples were taken from each patient and evaluated for detection of anti-Fasciola antibodies by an indirect ELISA. Patients who defined as having fascioliasis were treated with triclabendazole and were followed for at least three months for clinical improvement. Results: Serological test was positive in 5 patients. Of these 5 cases, three cases had a history of ingesting raw aquatic vegetables. The main clinical signs and symptoms in positive cases were; abdominal pain (60%), epigastric pain (40%), anemia (60%), and dermal pruritus (20%). Hypereosinophilia was seen in all of 5 positive cases. No Fasciola egg was found in stool specimens of any of the patients. The fascioliasis cases were treated by triclabendazole and clinical symptoms disappeared in all of 5 cases. Conclusion: Our observation further confirmed Yasuj district as a human endemic area for fascioliasis in Iran. The study also highlighted the importance of clinical features together with eosinophilia, as key parameters, in the diagnosis of human fascioliasis. Clinicians need to be aware of this disease and should keep in mind fascioliasis when hypereosinophilia present in patients in such endemic areas.

Toxoplasma gondii is zoonotic protozoa that spread throughout the world. The aim of current study was determination of the prevalence of T. gondii infection in patient undergoing chemotherapy for malignancies using ELISA and two different molecular tests including Nested-PCR and LAMP techniques. This study was carried out in Ali Ibn Aboutaleb hospital in Zahedan cities, southeast of Iran. Totally, 154 serum samples from patient were test for detection of IgG and IgM antibodies against T. gondii by ELISA test. All of the samples were also test by Loop amplification (LAMP) and Nested-PCR with RE gene. Totally, 39.4% of the sample sera from cases were positive for IgG against T. gondii. After using RE-Nested-PCR technique, 29.8% sample sera were positively. After using LAMP technique, 33.1% of the samples were found to be positive. Based on our result, the RE-LAMP technique was more effective than RE-nested-PCR method in the detection of T. gondii DNA.

Parasitological methods are not helpful for the diagnosis of fascioliasis in acute and invasive periods of the disease. Detection of coproantigens seems to be a suitable alternative approach in the diagnosis of fascioliasis. The present study aimed to develop a reliable antigen detection system, using monoclonal antibodies raised against excretory-secretory (ES) antigen of Fasciola hepatica, for the diagnosis of fascioliasis. Fasciola adult worms were collected from the bile ducts of infected animals. Species of the fluke was determined by polymerase chain reaction-restriction fragment length polymorphism (RFLP-PCR). ES antigen of F. hepatica was prepared. For production of monoclonal antibodies, mice were immunized with ES antigens of F. hepatica. Spleen cells from the immunized mice were fused with NS-1 myeloma cells, using polyethylene glycol. Hybridoma cells secreting specific antibody were expanded and cloned by limiting dilution. Moreover, polyclonal antibody was produced against F. hepatica ES antigen in rabbits. A capture enzyme-linked immunosorbent assay (ELISA) system, using produced monoclonal antibody, was designed and stool samples of infected animals along with control samples were tested by the system. The capture ELISA detected the coproantigen in 27 of 30 (90%) parasitologically confirmed fascioliasis cases, while 4 of 39 (10.25%) samples infected with other parasitic infections showed a positive reaction in this system. No positive reactivity was found with healthy control samples. Accordingly, sensitivity of 90% and specificity of 94.2% were obtained for the capture ELISA system. The results were compared with those obtained with commercial BIO-X ELISA, and a very good (kappa = 0.9) agreement was found between the commercial kit and the developed capture ELISA. Findings of this study showed that the produced monoclonal antibody has appropriate performance for the detection of Fasciola coproantigen in stool samples and can be appropriately used for the diagnosis of fascioliasis.

Latex agglutination test (KAtex) has been used in the last two decades for the diagnosis of visceral leishmaniasis (VL) in different VL-endemic areas. Here, we present a meta-analysis of studies which evaluated the KAtex for the diagnosis of VL to find out its overall diagnostic performance. A database search was performed on PubMed, Scopus, ISI Web of Science, Iranmedex and Google Scholar. The search of databases found 57 papers, of which 17 articles fulfilled our eligibility criteria. Meta-analysis of diagnostic accuracy (MADA) and Hierarchical Summary Receiver Operating Curve (HSROC) packages were used to do the meta-analysis and to obtain pooled estimates of sensitivity and specificity. Fixed effect bivariate analysis was conducted, using Mantel-Haenszel estimator, to measure the performance and diagnosis odds ratio (DOR) of the test. Heterogeneity of the test results was assessed by Chi-squared test. The sensitivity of individual studies ranged from 39.8 to 100%, and the specificity ranged from 64 to100%. The combined sensitivity and specificity estimates of KAtex were 77% (95% CI, 70-83%), and 97% (95% CI, 93-97%), respectively. Comparing the performance of the test by region suggests a significant difference where the lowest and highest sensitivities are reported from Nepal/Tunisia and Europe/Middle East respectively (p < 0.05). On the other hand, the lowest and highest rates of specificity were reported from Sudan and America/Middle East respectively. The overall specificity of KAtex is satisfactory. However, KAtex suffers from low sensitivity and this shortcoming should be improved. The test provides a rapid and simple diagnosis of VL and improvement of its sensitivity deserve further studies.

Background: Rodents are considered as reservoirs of various zoonotic diseases including helminthic infections. The current study aimed to evaluate the prevalence of helminth infections in rodents, in Boyer-Ahmad district, Southwestern Iran. Methods: Overall, 52 rodents were captured from various areas of the district by Sherman live traps. The animals were then euthanized and dissected. During necropsy, each organ was examined macroscopically for presence of any cyst or visible parasite. The gastrointestinal tract was removed and their contents were evaluated for larva or adult worms. Trichinella larvae in the rodents’ muscles were investigated by both digestion and pathological methods. Results: Twenty-eight (53.8%) of the trapped rodents were male. The rodents were including 25 (48.1%) Meriones persicus, 1(1.9%) Calomyscus bailwardi, 1 (1.9%) Arvicola terresterris, 7 (13.5%) Rattus rattus, 8 (15.4%) R. norvegicus, and 10 (19.2%) Apodemus sylvaticus. Of them, 38 (73.0%) were infected with at least one helminth. Collected rodents were infected with Hymenolepis diminuta (50%), Hymenolepis nana fraterna (28.8%), Skrjabinotaenia sp. (15.4%), Anoplocephalidae sp. (15.4%), Cysticercus fasciolaris (5.8%), Trichuris muris (36.5%), Aspiculuris tetraptera (15.4%), Syphacia sp. (5.7%), Rictularia sp. (15.4%), Trichostrongylus sp. (3.8%), and Gongylonema sp. (3.8%). M. persicus was the most (84%) infected rodent, yet the differences between rodent genus and helminth infectivity were not statistically significant (P>0.05). Conclusion: The rodents in Boyer-Ahmad district are infected with different helminths infections that some of them are recognized as threat to human health.

Serological assays have been extensively evaluated for diagnosis of visceral leishmaniasis (VL) and considered as a routine method for diagnosis of VL while these methods are not properly evaluated for diagnosis of cutaneous leishmaniasis (CL). This study aimed to assess the performance of indirect immunofluorescent-antibody test (IFA) and enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of cutaneous leishmaniasis in Iran. Sixty-one sera samples from parasitologically confirmed CL patients and 50 sera from healthy controls along with 50 sera from non-CL patients were collected. Antigen was prepared from promastigotes and amastigotes ofLeishmania major. IFA was used to detect anti-LeishmaniaIgG while ELISA was used to detect anti-LeishmaniaIgM, total IgG, or IgG subclasses (IgG1 and 4). ELISA, for detection of total IgG and IgM, showed sensitivity of 83.6% and 84.7% and specificity of 62.7% and 54.6%, respectively. Sensitivity and specificity of ELISA for detecting IgG1 and IgG4 were 64%, 75% and 85%, 49%, respectively. Sensitivity and specificity of IFA were 91.6% and 81%.Conclusion. Findings of this study demonstrated that serological test, especially IFA, can be used for proper diagnosis of CL.

Background: Toxoplasma gondii is a cosmopolitan protozoan, which infects the human body and many warm-blooded animals including wild boars. Wild boars have an important role in maintenance and transmission of T. gondii and may pose a potential risk to humans and wildlife health. Objectives: The current study was conducted to find out the seroprevalence and genotypes of T. gondii in wild boars in southwest of Iran. Methods: Twenty five adult wild boars were hunted, in 2013. Blood and tissue samples (muscle, tongue, liver, and brain) were collected from each animal. Blood samples were assessed for antibodies against T. gondii by the modified agglutination test. DNA was extracted from the tissues and PCR-amplified, targeting a 529 bp DNA fragment of T. gondii. The PCR products were purified and sequenced. Results: Anti-T. gondii antibodies were detected in sera of 4 out of the 25 (16%) wild boars. There was no gender or age-related differences in seroprevalence of T. gondii in the studied animals. PCR detected the 529 bp band of T. gondii in brain samples of 5 out of 25 (20%) of animals. Molecular evaluation confirmed the presence of type III (4 cases) and type I (1 case) of T. gondii in the studied wild boars. Conclusions: Findings of the study demonstrated that the T. gondii infection is common in wild boars in southwest Iran and could represent a significant health risk for animals, hunters, and local residents who may consume wild boar meat.

This study presents the first molecular and serological evaluation of Echinococcus granulosus infections in wild boars in Iran. Twenty five wild boars were collected in south-western Iran, during authorized hunting program, from March to October 2013, necropsied and examined for E. granulosus infection. Furthermore, seroprevalence of cystic echinococcosis in hunted boars was evaluated by an ELISA system. A fertile hydatid cyst due to E. granulosus was detected in the lung of one of the animals. Genotype analysis of the isolate was determined by analyzing a mitochondrial gene, cytochrome C oxidase subunit 1 (co1). DNA was extracted from the cyst sample and polymerase chain reaction amplification and DNA sequencing of the specific region of the co1 gene was performed. Molecular evaluation confirmed the presence of a sheep strain, the G1 genotype, in the wild boar in south-western Iran. This is the first report of the presence of G1 genotype of E. granulosus in wild boar in Iran. Serological evaluation of hydatid cyst by antigen-B ELISA revealed E. granulosus antibodies in 5 (20%) of 25 wild boars. A statistically significant difference was observed between the prevalence of E. granulosus antibodies and gender while the difference between the seroprevalence of E. granulosus and age was insignificant. Findings of this study might have important implications for the prevention and control of cystic echinococcosis.

Background: Cutaneous leishmaniasis (CL) is a parasitic disease with a relatively wide distribution in different areas of the world, including Iran. The parasite is mainly diagnosed microscopically, but serological approaches might be useful for diagnosis as well. This study aimed to assess the efficacy of an immunoblotting system for serodiagnosis of cutaneous leishmaniasis in Iran. Methods: Sixty-one sera samples from parasitologically confirmed CL patients and 50 sera samples from healthy controls along with 50 sera sample from non-CL patients were collected. Native strain of Leishmania major was cultured in Schneider medium and soluble Leishmania antigens were prepared from amastigotes-like parasites. All of sera samples were evaluated by an immunoblotting system. Results: Components of 14 to 135 kDa were detectable by the sera of CL patients. From 61 sera of CL patients, 59 cases (96.7%) detected a 63 kDa subunit and 51 cases (83.6%) recognized a 32–35 kDa component. Among all subunits, the 63 kDa band showed the highest sensitivity (96.7%) and a 75 kDa band had the highest (98%) specificity. Conclusion: Immunoblotting has a satisfactory performance in diagnosis of CL and this test can be used, as an aid, for proper diagnosis of CL.

Background: Toxoplasmosis is a life-threatening infection in organ transplant recipients, people receiving corticosteroid or radiation therapy, people with malignancies, and AIDS patients. Objectives: The current study aimed to determine the prevalence of toxoplasmosis in patients receiving chemotherapy for malignancies in the Bushehr province of southwest Iran. Methods: Blood samples were taken from 86 patients who were continuously referred to the chemotherapy center in Bushehr province and evaluated by ELISA to determine anti-Toxoplasma IgG and IgM antibodies. Moreover, a blood buffy coat of each sample was assessed by polymerase chain reaction (PCR), targeting a 529 bp gene of T. gondii. PCR products of the positive samples were sequenced to determine the genotype of the parasite. Results: Anti-Toxoplasma IgG antibodies were detected in the sera of 21 (24.4%) cases. All of the patients were negative for anti-Toxoplasma IgM antibodies. No statistically significant correlation was found between seropositivity to Toxoplasma and duration of chemotherapy or having contact with cats. PCR detected a 529 bp band of T. gondii in the buffy coats of two out of 86 (2.3%) cases. The sequence analysis demonstrated that both cases were 95% identical to type III (VEG strain) of T. gondii. Conclusions: Findings of this study demonstrated the presence of type III T. gondii in the buffy coats of patients undergoing chemotherapy. Given that toxoplasmosis is a life-threatening infection in immunocompromised patients, these patients should be screened for toxoplasmosis before and during chemotherapy to prevent acute toxoplasmosis.

We report a fatal congenital toxoplasmosis case in an Iranian woman in the south of Iran. A pregnant mother had been admitted at the 15th week of her pregnancy on account of a febrile illness, symptoms of common cold, and enlargement of submandibular lymph nodes. Serological testing of the mother’s serum revealed positive IgG and IgM anti-Toxoplasmaantibodies. Amniotic fluid was taken and evaluated by polymerase chain reaction (PCR) assay with a direct amplification of theToxoplasmaURPT gene which was found to be positive. Sequencing and analysis of PCR product revealed that the isolate has the most similarity with type I ofToxoplasma gondii. Fetal scan showed anomaly in fetus including mild hydrocephaly. Termination of the pregnancy was suggested by the physician and pregnancy was terminated 178 days after conception.

Background: Seroprevalence of toxoplasmosis in different populations in Iran varies according to the people eating behaviors and also geographic and climatic differences of each area. Objective: The current study aimed to provide recent data regarding the seroprevalence of toxoplasmosis among female university students in Shiraz University of Medical Sciences, in Fars province, southern Iran. Materials and Methods: The subjects of the study were 503 female university students. Blood samples were collected from each participant and tested for anti-Toxoplasma IgG and IgM antibodies, using a commercial Enzyme-Linked Immunosorbent Assays (ELISA) kit. Demographic characteristics and risk factor related to Toxoplasma were also recorded during the samples collection. Results: The mean age of participants was 22.2 (±3.83) years and the majority (54.9%) of subjects was in the age group of 20-25 years old. Anti T. gondii antibodies was detected in sera of 43 out of 503 enrolled students, corresponding to an overall seroprevalence of 8.5%. Of these, 37 (7.4%) were seropositive for only IgG, 7 (1.4%) were seropositive for only IgM and 1 (0.2%) were seropositive for both IgG and IgM. The differences between age and animal contact with seropositivity to toxoplasmosis were not statistically significant. Conclusion: The findings of the study indicated that more than 90 % of the female university students in this study were seronegative for toxoplasmosis. As these students are in their childbearing age, there is a possibility for their newborns to become infected with Toxoplasma. The control and preventative measurements are necessary to reduce the rate of T. gondii infection in such individuals.

Visceral Leishmaniasis (VL) is an endemic parasitic disease and remains as a major health concern in southwestern Iran. The current study describes clinico-hematological, epidemiological and therapeutic features of VL cases, admitted to university-affiliated hospitals, during 1999-2014 in Fars province, southwestern Iran. A total of 380 VL cases were recorded during a 16 years period, giving an average annual admission of 23.75 cases/year in which 217 (57.1%) were male and 163 (42.9%) were female. Mean age of the patients was 3.7 years. The majority of the cases (91.5%) were ≤ 5 years old. Bone-marrow aspiration detected Leishmania amastigotes only in 26.6% of cases. Fever (98.1%), abdominal protrusion (65.1%) and hepatosplenomegaly (63.7%) were the most common clinical presentations of the patients. Pancytopenia was noted in 43.1, anemia in 87.3 and thrombocytopenia in 64% of cases. Increase in the level of AST (aspartate aminotransferase), ALT (alanine aminotransferase), alkaline phosphatase, LDH (lactate dehydrogenase) and CRP (C-Reactive Proteins) were seen in 84.9, 53.6, 44.4, 72.5 and 83.1% of cases, respectively. Mortality was noted in 5.3% of cases. Deranged haemato-biochemical parameters including total and direct bilirubin, PLT (platelet) and pancytopenia were significantly contributed to mortality from VL. Moreover, clinical features such as severe splenomegaly as well as bacterial infections were meaningfully contributed to death from VL. The majority of patients (74.9%) were treated with meglumine antimoniate. Amphotericin B was administrated in 59 of cases, 11 of them were initially treated with meglumine antimoniate with a shift to amphotericin B, because of treatment failure. Findings of the current study demonstrated that VL is present in southwest of Iran with a fairly continual rate during the last 16 years period. Deranged haemato-biochemical parameters along with severe splenomegaly contributed to mortality from VL.