Diagnostic study for Toxoplasmosis by Serological test and PCR technique in Kirkuk province
Article Sidebar
Main Article Content
Abstract
Background: Severe Toxoplasma gondii infection in early pregnancy brings the risk of transferring the infection to the unborn baby with serious abnormality. The aim of identifying toxoplasmosis in early stage is to recognize mother-to-infant transmission for early cure to avoid undesirable complication. Using serological test to determine anti-Toxoplasma specific IgG and IgM antibodies possibly may not identify new or even previous infection. However, many molecular techniques are now being extensively used through the world among these methods polymerase chain reaction PCR technique emerged as the most widely accepted method for directly detection infectious agents. Thus, the goal in this study is to compare immunoassay by indirect (IgM and IgG) detection diagnostic technique and conventional PCR as molecular assay in diagnosis of T. gondii in whole blood of aborted females in Kirkuk province-Iraq.
Methods: Here we applied indirect routine serological diagnosis test for detection toxoplasma specific IgM and IgG antibodies in females blood with spontaneous abortion and compare the efficiency of this method with PCR technique to detect the pathogenic protozoan T. gondii based on identification of (the B1 gene) as a target. Eighty-one females were incorporated in this study with a history of recurrent spontaneous abortions. All of these cases were limited to only females in the reproductive age (19-44 years). There are several serological diagnostic test kits available in Iraqi markets; however, the sensitivity and effectiveness of these commercial kits are not examined by most of the laboratories before they are obtained. Therefore, blood samples from female patients with single or repeated abortion were tested for detection specific anti-Toxoplasma antibodies using commercially available (Medical-Biozec) kit in Kirkuk province and compare that with the results are obtained from conventional PCR assay.
Results: The results showed that the conventional-PCR analysis is remarkably more sensitive than serological test, with a relatively high proportion of the positive samples for PCR compared with the serological diagnosis. The serological test results revealed that anti-T. gondii antibodies were detected in 14.81% of aborted women, 3.7% of them were positive for IgM, while 11.11% were positive for IgG. On the contrary, conventional PCR analysis found that 44.44% of aborted women showed positive result for B1 gene of T. gondii. Despite this, most of positive serology-aborted women exposed positive result for B1 gene of T. gondii, except two cases include false-positive IgM results
Article Details
This work is licensed under a Creative Commons Attribution 4.0 International License.
Tikrit Journal of Pure Science is licensed under the Creative Commons Attribution 4.0 International License, which allows users to copy, create extracts, abstracts, and new works from the article, alter and revise the article, and make commercial use of the article (including reuse and/or resale of the article by commercial entities), provided the user gives appropriate credit (with a link to the formal publication through the relevant DOI), provides a link to the license, indicates if changes were made, and the licensor is not represented as endorsing the use made of the work. The authors hold the copyright for their published work on the Tikrit J. Pure Sci. website, while Tikrit J. Pure Sci. is responsible for appreciate citation of their work, which is released under CC-BY-4.0, enabling the unrestricted use, distribution, and reproduction of an article in any medium, provided that the original work is properly cited.
References
1. Ferguson DJP. Toxoplasma gondii: 1908-2008, homage to Nicolle, Manceaux and Splendore. Memórias do Instituto Oswaldo Cruz. 2009;104:133-148.
2. Wong S-Y, Remington JS. Toxoplasmosis in pregnancy. Clinical Infectious Diseases. 1994:853-861.
3. Van de Ven E, Melchers W, Galama J, Camps W, Meuwissen J. Identification of Toxoplasma gondii infections by BI gene amplification. Journal of clinical microbiology. 1991;29(10):2120-2124.
4. Lin M-H, Chen T-C, Kuo T-t, Tseng C-C, Tseng C-P. Real-Time PCR for Quantitative Detection ofToxoplasma gondii. Journal of clinical microbiology. 2000;38(11):4121-4125.
5. Wiengcharoen JT, Chiabchalard R, Sukthana Y. PCR technique for detecting Toxoplasma gondii in animal amniotic fluid. The Southeast Asian Journal of Tropical Medicine and Public Health 2004;35(4): 792-5.
6. Singh S. Mother-to-child transmission and diagnosis of Toxoplasma gondii infection during pregnancy. Indian journal of medical microbiology. 2003;21(2):69.
7. James GS, Sintchenko VG, Dickeson DJ, Gilbert GL. Comparison of cell culture, mouse inoculation, and PCR for detection of Toxoplasma gondii: effects of storage conditions on sensitivity. Journal of clinical microbiology. 1996;34(6):1572-1575.
8. Ho-Yen D, Joss A, Balfour A, Smyth E, Baird D, Chatterton J. Use of the polymerase chain reaction to detect Toxoplasma gondii in human blood samples. Journal of clinical pathology. 1992;45(10):910-913.
9. Romand S, Wallon M, Franck J, Thulliez P, Peyron F, Dumon H. Prenatal diagnosis using polymerase chain reaction on amniotic fluid for congenital toxoplasmosis. Obstetrics & Gynecology. 2001;97(2):296-300.
10. Bartlett JMS, Stirling D. PCR protocols: method in molecular biology. Human Press. 2003;226(2nd edi).
11. Al-Sanjary R, Hussein T. Using species-specific PCR technique to detect Toxoplasma gondii in broiler chickens. Iraqi Journal of Veterinary Sciences. 2012;26(2):53-56.
12. Montoya JG. Laboratory diagnosis of Toxoplasma gondii infection and toxoplasmosis. Journal of Infectious Diseases. 2002;185(Supplement 1):S73-S82.
13. Wallon M, Franck J, Thulliez P, Huissoud C, Peyron F, Garcia-Meric P, et al. Accuracy of real-time polymerase chain reaction for Toxoplasma gondii in amniotic fluid. Obstetrics & Gynecology. 2010;115(4):727-733.
14. Said RN, Zaki MM, Abdelrazik MB. Congenital toxoplasmosis: evaluation of molecular and serological methods for achieving economic and early diagnosis among Egyptian preterm infants. Journal of Tropical Pediatrics. 2010.
15. Salman YJ. Watching of Toxoplasma gondii antibodies among peoples in Kirkuk Province from 1993 to 2012 by using different serological tests. Int J Curr Microbiol App Sci. 2014;3(9):923-932.
16. Kasim TN. Efficacy of some laboratory methods in detecting protozoan parasites in Kirkuk city. Msc thesis College of Science, Kirkuk University. 2013.
17. Tewfik SK. Some immunological aspects of toxoplasmosis among women with abortion and congenital abnormalities. MSc thesis College of Medicine-Tikrit University. 2013.
18. Gras L, Gilbert R, Wallon M, Peyron F, Cortina-Borja M. Duration of the IgM response in women acquiring Toxoplasma gondii during pregnancy: implications for clinical practice and cross-sectional incidence studies. Epidemiology and infection. 2004;132(03):541-548.
19. Liesenfeld O, Press C, Montoya JG, Gill R, Isaac-Renton JL, Hedman K, et al. False-positive results in immunoglobulin M (IgM) toxoplasma antibody tests and importance of confirmatory testing: the Platelia Toxo IgM test. Journal of clinical microbiology. 1997;35(1):174-178.
20. Said RN, Zaki MM, Abdelrazik MB. Congenital toxoplasmosis: evaluation of molecular and serological methods for achieving economic and early diagnosis among Egyptian preterm infants. Journal of tropical pediatrics. 2010:fmq097.
21. Tlamçani Z, Lemkhenete Z, Lmimouni BE. Toxoplasmosis: The value of molecular methods in diagnosis compared to conventional methods. Journal of Microbiology and Infectious Diseases. 2013;3(02).
22. Wastling JM, Nicoll S, Buxton D. Comparison of two gene amplification methods for the detection of Toxoplasma gondii in experimentally infected sheep. Journal of Medical Microbiology. 1993;38(5):360-365.
23. Brézin AP, Egwuagu CE, Burnier M, Silveira C, Mahdi RM, Gazzinelli RT, et al. Identification of Toxoplasma gondii in paraffin-embedded sections by the polymerase chain reaction. American journal of ophthalmology. 1990;110(6):599-604.
24. Jones CD, Okhravi N, Adamson P, Tasker S, Lightman S. Comparison of PCR detection methods for B1, P30, and 18S rDNA genes of T. gondii in aqueous humor. Investigative ophthalmology & visual science. 2000;41(3):634-644.
25. Tekkesin N. Diagnosis of toxoplasmosis in pregnancy: a review. HOAJ Biology. 2012;1(1):9.
26. Vieira dos Santos MdS, Lima da Silva CG, Lima de Oliveira PN, Barros Ribeiro KD, Teixeira Júnior AG, Aguiar Santos MF, et al. Diagnosing congenital toxoplasmosis: where are we? A systematic review. International Archives of Medicine; Vol 8 (2015)DO - 103823/1656. 2015.
27. Ashraf M, Abdul-Haleem S. Seroprevalence Of Anti Toxoplasma gondii IgG and IgM Among Pregnant Women in Sana’a Capital and Capital Trusteeship. Scientific Journal of King Faisal University (Basic and applied Sciences). 2010;11(2): 1431.
28. Sakikawa M, Noda S, Hanaoka M, Nakayama H, Hojo S, Kakinoki S, et al. Anti-Toxoplasma antibody prevalence, primary infection rate, and risk factors in a study of toxoplasmosis in 4,466 pregnant women in Japan. Clinical and Vaccine Immunology. 2012;19 (3):365-367.
29. Delpisheh A, Brabin L, Attia E, Brabin BJ. Pregnancy late in life: a hospital-based study of birth outcomes. Journal of women's health. 2008;17 (6):965-970.