Serological and molecular approaches in clinical diagnosis of ocular toxoplasmosis in Iran.

Background: Toxoplasma gondii (T. gondii) is the most common parasite that can lead to a disease called toxoplasmosis. In this study, serological and molecular complementary tests have been conducted to detect or diagnose this parasite. Methods: A total of 71 patients with clinical symptoms of ocular toxoplasmosis and 20 patients with other ocular infections were evaluated. Serum and buffy coat samples were collected and tested using enzyme-linked immunosorbent assay (ELISA) and nested polymerase chain reaction (nPCR) assessments. Superficial T. gondii B1 gene was evaluated in PCR. The ocular toxoplasmosis patients were followed-up 2 weeks after the first sampling and 4 weeks following the first laboratory testing. The main outcome measures were the efficiency of the diagnostic procedure and positive and negative predictive values (PPV and NPV). Results: Overall, of the samples, 69% were PCR+, IgG+, and IgM-, and 4.2% showed PCR+, IgG+, and IgM+. In the first follow-up, after 2 weeks, from the 41 referred patients, 29 (70%) showed PCR+, IgG+, and IgM-, which confirmed the results of the first sampling. In the second follow-up, 9 (47%) patients were PCR+, IgG+, and IgM-. A correlation was observed between the first referral and the follow-ups. Also, from 71 patients, diagnosed clinically as ocular toxoplasmosis, the disease was confirmed in 73.2% and 26.8% of those suffering from other ocular infections. Of the 20 control group samples, 55% showed PCR-, IgG+, and IgM-. The sensitivity, specificity, negative and positive predictive values, and negative and positive likelihoods were analyzed for IgG and IgM antibodies and for PCR using ELISA method. Conclusion: As the ophthalmologic signs of T. gondii may be mimicked by other infections, clinical methods may be complemented by laboratory approaches for a definite diagnosis. This would assist clinicians to achieve timely diagnosis and successful therapy and to control the infection.

Serological and Molecular Diagnosis of Toxoplasma gondii Infections in Thalassemia Patients.

BACKGROUND: This study aimed to the serological and molecular diagnosis of Toxoplasma gondii infections and related risk factors in patients with thalassemia major and healthy controls. METHODS: This case-control study was performed in Shahrekord University of Medical Sciences, Shahrekord, west of Iran from Jan 2014 to Jan 2015. Overall, 235 patients with thalassemia major and 235 healthy controls were enrolled. Assessment of anti-Toxoplasma antibodies in sera samples was performed using commercial ELISA kits. In order to the molecular investigate of T. gondii in blood samples, a relatively new molecular assay, LAMP technique based on Toxoplasma SAG1 gene was conducted for the first time. The specificity of LAMP outer primers for the T. gondii detection was confirmed by sequencing the purified PCR product. RESULTS: 51.9% of thalassemia patients and 34.8% of healthy controls were positive for anti-Toxoplasma IgG antibodies, which the difference was statistically significant (P<0.01). In terms of anti-Toxoplasma IgM antibody, 3.4% of thalassemia patients and 2.1% of healthy individuals were positive, which the difference was not statistically significant (P=1). Based on SAG1-LAMP, 9.78% of the thalassemia patients and 5.95% of healthy controls were positive for T. gondii DNA, which the difference was not statistically significant (P≤0.230). CONCLUSION: Thalassemia patients, probably due to repeated blood transfusion and consequently, immune deficiency, are at risk of transmitting Toxoplasma infection more than healthy people. Therefore, screening of Toxoplasma infection in blood transfusion centers may be effective in the prevention of toxoplasmosis in these patients.

An updated meta-analysis of the association between Toxoplasma gondii infection and risk of epilepsy.

Toxoplasma gondii is a neurotropic pathogen with worldwide distribution. To evaluate the association between Toxoplasma infection and the risk of epilepsy by meta-analysis, observational peer-reviewed studies were retrieved from PubMed, Embase, Web of Science, Scopus and Google Scholar (up to 10 October 2018) and by reference review. Pooled risk estimates were calculated using a random effects model. Heterogeneity was assessed using Cochrane's Q-test and I2. In total, 16 eligible studies involving 19 data sets were included for the final analysis. A total 7897 participants (3771 epileptic patients, 4026 healthy controls) were included. The pooled odds ratio (OR) for Toxoplasma infection was increased to 1.72 (95% confidence interval [CI] 1.37 to 2.16) among patients with epilepsy. There was moderate heterogeneity among the studies (χ2=39.8, I2=62.3%, p=0.001). The ORs from subgroup analyses showed that both cryptogenic epilepsy (OR 2.65 [95% CI 1.91 to 3.68]) and active convulsive epilepsy (OR 1.37 [95% CI 1.09 to 1.72]) were significantly associated with Toxoplasma infection. Another subgroup analyses according to age showed a significant positive association in children (OR 1.33), adults (OR 1.57) and in all ages (OR 1.89). Our findings support the association between Toxoplasma infection and epilepsy. More prospective studies with larger sample sizes and more experimental studies are recommended to elucidate a causative relationship.

Helicobacter pylori infection and risk of preeclampsia: a systematic review and meta-analysis.

OBJECTIVE: Helicobacter pylori is associated with many pregnancy adverse effects such as preeclampsia (PE). We performed this systematic review and meta-analysis study to assess the possible association between H. pylori infection and PE and this is the first meta-analysis to clarify this issue. METHODS: PubMed, ISI (Web of Science), SCOPUS, and Google Scholar databases were searched (up to April 2017) to identify the relevant studies. The Meta-analysis of Observational Studies in Epidemiology and Preferred Reporting Items for Systematic Reviews and Meta-Analyses Protocols guidelines were used to do this study. Pooled odds ratio (OR) and 95% confidence intervals (CI) were estimated using a random-effects meta-analysis model. Heterogeneity was assessed with the χ2-based Q-test and I2 statistic. RESULTS: A total of eight studies including 889 participants (460 preeclamptic women and 429 controls) met the eligibility criteria. A positive association was found between H. pylori infection and PE (OR: 3.35; 95% CI: 2.21-5.10). Heterogeneity was acceptable (χ2 = 13.39; I2 = 47.7, 95% CI: 0-77). In subgroup analysis, cytotoxin-associated antigen A seropositivity was a substantial risk factor for PE when immunoblotting methods (OR: 11.12; 95% CI: 5.34-23.16; χ2 = 6.42; I2 = 53.3, 95% CI: 0-85) were used, whereas it was not potential risk factor for PE when ELISA was used as a detecting method (OR: 1.11; 95% CI: 0.6-2.06; χ2 = 1.83; I2 = 0, 95% CI: 0-90). CONCLUSIONS: This study indicated that women with H. pylori infection, especially those infected with Cag A positive strains are more likely to have PE compared with the uninfected women.

Seroepidemiology, modifiable risk factors and clinical symptoms of Toxocara spp. infection in northern Iran.

Toxocariasis is one of the most important and widespread neglected tropical infectious diseases. We designed a cross-sectional study to assess the seroepidemiological aspects of toxocariasis among the general population in northern Iran. A total of 630 rural subjects were enrolled to participate in the study. The presence of anti-Toxocara immunoglobulin G (IgG) was tested using a commercially available enzyme-linked immunosorbent assay. A structured questionnaire was also used to evaluate the potential risk factors and related clinical signs/symptoms. The presence of anti-Toxocara IgG antibodies was detected in 148 of the 630 rural subjects (23.5% [95% confidence interval {CI} 21.8 to 25.1]). By multivariate analysis, age (odds ratio [OR] 2.89 [95% CI 1 to -8.3], p=0.04), eating improperly washed vegetables (OR 4.05 [95% CI 2.47 to 6.64], p<0.001), contact with dogs (OR 3.31 [95% CI 2.13 to 5.12], p<0.001) and exposure to soil (OR 3.56 [95% CI 2.13 to 5.13], p<0.001) were significantly associated with the seroprevalence of Toxocara. The clinical study also demonstrated that the seroprevalence of Toxocara infection was significantly associated with asthma (OR 3.78 [95% CI 1.63 to 8.75], p<0.001) and ophthalmic disorder (OR 1.83 [95% CI 1.04 to 3.21], p=0.034). The findings proved that residents of rural communities in tropical regions may be heavily exposed to Toxocara spp. We highly recommend more investigations in high-risk groups.

The relationship between Toxocara species seropositivity and allergic skin disorders: a systematic review and meta-analysis.

We performed a systematic review and meta-analysis on observational studies to evaluate the possible associations between Toxocara species seropositivity and allergic skin disorders (ASDs). We searched the MEDLINE, ScienceDirect, Scopus, Web of Science and Google Scholar databases to 15 May 2018 to identify the relevant studies. We used a random effects meta-analysis model to generate the pooled odds ratio (OR) and 95% confidence intervals (CIs). Fifteen studies, including eight studies with a case-control design (735 patients and 1342 controls) and seven studies with a cross-sectional design (a total of 4804 participants, 1302 individuals with ASDs and 3502 without ASDs), were included in the meta-analysis. We found an increased risk for ASDs in individuals with Toxocara seropositivity (OR 1.75 [95% CI 1.16 to 2.64]). Subanalysis showed that Toxocara seropositivity was significantly associated with urticaria (OR 2.97 [95% CI 1.53 to 5.76]), however, it was not significantly associated with atopy (OR 1.08 [95% CI 0.55 to 2.15]) and eczema (OR 1.62 [95% CI 0.95 to 2.78]). Moreover, the pooled ORs were 2.34 (95% CI 1.32 to 4.15) and 1.27 (95% CI 0.69 to 2.35) for case-control and cross-sectional studies, respectively. The results of our study support hypotheses regarding the existence of a positive relationship between Toxocara infection and allergic disorders, although this association should be further investigated by longitudinal and mechanism studies.

The global seroprevalence of Toxoplasma gondii among wild boars: A systematic review and meta-analysis.

This systematic review and meta-analysis study was performed to evaluate the worldwide seroprevalence of Toxoplasma gondii among wild boar. We searched PubMed, Science Direct, Web of Science, Cochrane, Scopus, EBSCOhost and Google Scholar databases for studies reporting T. gondii seroprevalence in wild boars between January 1995 and March 2017. Inclusion and exclusion criteria were applied. We estimated the pooled seroprevalence of T. gondii in wild boars using a random-effects model, and evaluated overall seroprevalence in different geographical areas. A total of 43 articles that included 16788 wild boar from 23 countries fulfilled our eligibility criteria. Of these, 4759 wild boar had been defined T. gondii seropositive and we estimated the pooled worldwide seroprevalence of toxoplasmosis in wild boars to be 23% (95% CI: 19-27%). The pooled seroprevalence in North America (32%, 20-45%; odds ratio [OR] 2.09) and Europe (26%, 21-30%; OR 1.72), was higher than Asia (13%, 5-23%). The lowest seroprevalence was estimated in South America (5%, 3-8%). An increased seropositivity was observed with elevation in geographical latitude. In subgroup analyses, the pooled seroprevalence of T. gondii was higher in wild boar older than 12 months of age (28%, 22-35%; OR 1.57) compared to those up to 12 months of age (20%, 16-25%). Our findings suggest that wild boar have an important role in human infection and the epidemiological cycle of T. gondii infection.

The role of Blastocystis sp. and Dientamoeba fragilis in irritable bowel syndrome: a systematic review and meta-analysis.

Irritable bowel syndrome (IBS) is globally one of the most prevalent gastrointestinal disorders with a negative impact on quality of life and socio-economic status of patients. Recently, controversial evidences suggest that Blastocystis sp. and Dientamoeba fragilis infections may be implicated in the development of IBS. We performed a systematic review and meta-analysis to examine the possible association regarding this issue. PubMed, ScienceDirect, Scopus, Web of Science, and Cochrane electronic databases were searched (up to February 2017) to identify the relevant studies. Pooled odds ratio (OR) and 95% confidence intervals were estimated using a random effects meta-analysis model on data from included studies. A total of 17 studies including 5882 participants (2527 patients and 3310 controls) met the eligibility criteria. Individuals with Blastocystis infection were found to have a positive association with IBS (OR, 2.19; 95% CI, 1.54-3.13), while this association was not observed for D. fragilis infection (OR, 1.13; 95% CI, 0.22-5.72). In subgroup analysis for Blastocystis infection, the pooled ORs were OR 2.29, 95% CI 1.55-3.41; OR 1.70, 95% CI 0.83-3.44; and OR 3.83, 95% CI 2.34-6.27 for hospital-based, healthy volunteers, and combined controls, respectively. Considering the subtypes, meta-analysis result demonstrated significant positive ORs for ST1 (OR, 4.40; 95% CI, 2.81-6.90) and ST3 (OR, 1.94; 95% CI, 1.36-2.77) to be potential risk factors for IBS. Our results support the existence of a positive association between Blastocystis sp. and IBS. Further studies with more sample size should be performed to better investigate the real impact of these parasites on the occurrence of IBS.

Human infectious diseases and risk of preeclampsia: an updated review of the literature.

BACKGROUND: Preeclampsia (PE) is one of the major causes of maternal and perinatal morbidity and mortality, especially in low- and middle-income countries. In recent years, a growing body of literatures suggests that infections by bacteria, viruses, and parasites and their related inflammations play an important role in the pathogenesis of PE. METHODS: We searched PubMed, Google scholar, and Cochrane databases using the following search words: "infection and preeclampsia," "bacterial infection and preeclampsia," "viral infection and preeclampsia" and "parasitic infection and preeclampsia." RESULTS: The literature review revealed that many bacteria including Helicobacter pylori, Chlamydia pneumonia, and those are involved in periodontal disease or urinary tract infections (UTIs) and some viral agents such as Cytomegalovirus, herpes simplex virus type-2, human immunodeficiency virus, and some parasites especially Plasmodium spp. and Toxoplasma gondii can be effective in development of PE. Inflammation responses against infections has major role in the inducement of PE. The shift of immunological cytokine profile of Th2 toward Th1 and high levels of pro-inflammatory cytokines (TNF-ɑ, IL-12, IFN-γ, etc.), increase of oxidative stress, increase of anti-angiogenic proteins, increase of vascular endothelial growth factor receptor 1 (sVEGFR1), and complement C5a are the main potential mechanisms related to infections and enhanced development of PE. CONCLUSION: Thus, early diagnosis and treatment of bacterial, viral, and parasitic infections could be an effective strategy to reduce the incidence of PE.

Genotyping of Toxoplasma gondii Strains Isolated from Patients with Ocular Toxoplasmosis in Iran.

BACKGROUND: Toxoplasma gondii is an obligate, intracellular protozoon that develops its sexual stage in cat's intestinal epithelial cells as definitive host and develops its asexual stage in different tissues of a wide range hosts called intermediate host. The protozoon is a food-borne and worldwide parasite that can cause serious complications such as abortion in pregnant women, encephalitis, and ocular toxoplasmosis. The present study aimed to genotype T. gondii strains isolated from patients with toxoplasmic retinochoroiditis. METHODS: Fifty-two blood samples were taken from patients with ocular toxoplasmosis, from July 2013 to July 2014. The specimens were collected from three ophthalmological hospitals of Tehran, Iran. After that, DNA extraction was performed using kit on separated buffy coats of serologically positive blood samples. Then PCR was done in GRA6 gene. For digestion of products, MseI endonuclease was used. Finally, some of the PCR products were sequenced. RESULTS: All of 52 samples were found positive by serological and PCR-RFLP methods and all of isolated strains belong to type III genotype. Type III genotype has the highest prevalence in Iranian ocular toxoplasmic patients. CONCLUSION: T. gondii, particularly its type III should not be neglected as a cause of retinochoroiditis.