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Diagnosis of Toxoplasmosis in Pregnant Women Using Enzyme Immunoassay and Nested Real Time PCR in Al-Najaf city/Iraq
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| Author:
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MUNA GHAZZAY, KHALIDA K ABBAS AL-KELABY, EMAD SADIQ ALI AL-HILLI, NAJAH R. HADI
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| Abstract:
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This work was planned as diagnostic study to evaluate the prevalence of Toxoplasma Gondi (T. Gondi) among the population of 146 pregnant women of ages ranging from 14-45 years old, during the period from 20th of October 2018 to 14th of January 2019 at Al Najaf Public Health laboratory, and 50 normal pregnant women (control group), these cases were distributed according to the age groups, residency and IgM or IgG seropositivity. The diagnostic assay was firstly depending on Enzyme immunoassay (EIA) for the evaluation the prevalence of anti- T. Gondi IgM and IgG antibodies in the sera of pregnant women, then confirmed by the molecular technique (Real-time PCR) using specific primers for the detection of B1 gene of T. Gondi. This detection was also followed by confirmatory nested polymerase chain reaction using RE gene. Results presented in this study showed that overall seroprevalence of toxoplasmosis were 55:146 (37.67%). Out of 146 women; 53(36.3%) of samples had chronic toxoplasmosis characterized by the presence of positive IgG antibodies, while only 2 (1.37%) were positive for acute toxoplasmosis characterized by the presence of positive IgM antibodies. The prevalence of abortion according to age groups revealed that 34.9% (51:146) of the total pregnant cases; 17:53 (32.08%) of chronic T. Gondi infection and the two positive acute cases (2:51) of T. Gondi infection were related to women of 20-25 years age group. While 30.19% of toxoplasmosis seropositivity (16:53) were related to 32-37 years age group. According to the residency, 86:146(58.91%) were from rural area, while 60:146 (41.09%) were from urban area in Al-Najaf. Results also showed that out of 55 T. Gondi seropositivity cases, 39 (70.9%) were living in urban area and 16 (29.01%) were living in rural area. About 71.69% of IgG seropositivity patients were lived in urban area and about 28.31% were lived in rural area. Data regarding the two IgM positive cases showed that the infection distributed evenly (50%) for both rural and urban areas in Al-Najaf. Results were also showed statistically non-significant differences in accordance to the hospitalization need Control group investigation showed that 7:50 (14%) cases were seropositivity for anti- T. Gondi IgG, while all control cases revealed IgM seronegativity. The statistical analysis of PCR data of T. Gondi threshold cycle (CT) of B1 and RE genes were revealing mean ±Standard deviation of 18.6± 5.2 and 13.1± 1.3 respectively. The interassay coefficient of variation (CV) of B1 and RE genes of T. Gondi was 28% and 10.3% respectively. We concluded that B1 and RE genes detection was successful, confirmatory to EIA and more précised for T. Gondi isolated from serum samples.
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| Keyword:
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Toxoplasmosis, B1 gene, RE sequence, Nested PCR.
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| EOI:
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| DOI:
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https://doi.org/10.31838/ijpr/2020.12.02.0022
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| Download:
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