DOI: https://doi.org/10.14710/jbtr.v5i2.4840

Nested PCR methods for detection Toxoplasma gondii B1 gene in Cerebrospinal Fluid of HIV patients

1Faculty of Medicine, Diponegoro University, Indonesia

2Universitas Sumatera Utara, Indonesia

3Universitas Indonesia, Indonesia

Received: 8 May 2019; Revised: 10 Oct 2019; Accepted: 10 Oct 2019; Available online: 31 Dec 2019; Published: 31 Dec 2019.
Open Access Copyright (c) 2019 Journal of Biomedicine and Translational Research

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Abstract

Background: Toxoplasmosis is a disease caused by infection of Toxoplasma gondii, Which may cause a life-threatening condition in immunocompromised patients, for example, Toxoplasma encephalitis (TE). It is challenging to diagnose Toxoplasma as a cause of central nervous system (CNS) infection in HIV patient, so we need an alternative method, which is a PCR detection of Toxoplasma gondii B1 gene.

Objective: This research aimed to find association between PCR methods for Toxoplasma gondii B1 gene and anti-Toxoplasma IgG from cerebral spinal fluid patient HIV AIDS.

Methods: A cross-sectional study was conducted to Cerebrospinal fluid (CSF) samples of HIV patients with neurological symptoms to determine Toxoplasma gondii infection using nested PCR methods for the B1 gene and detection of anti-Toxoplasma IgG.

Results: 88 CSF samples from HIV patients tested using nested PCR showed 23 samples (26,1%) were positive. Serologic test for IgG Toxoplasma showed 34 samples were positive (28,6%). There was a significant correlation (p=0.000(<0.05) between PCR result and a serologic test for IgG Toxoplasma.

Conclusion: Nested PCR methods to detect B1 gene increased the accuracy of diagnosis for toxoplasma encephalitis.

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Keywords: cerebrospinal fluid; Toxoplasma gondii; PCR; IgG anti Toxoplasma; HIV

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Section: Original Research Articles
Language : EN
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  1. Luma HN, Tchaleu BC, Mapoure YN, Temfack E, Doualla MS, Halle MP, et al. Toxoplasma encephalitis in HIV/AIDS patients admitted to the Douala general hospital between 2004 and 2009: a cross sectional study. BMC Res Notes. 2013;6(1):146
  2. Terazawa A, Muljono R, Susanto L, Margono SS, Konishi E. High Toxoplasma antibody prevalence among inhabitants in Jakarta, Indonesia. Jpn J Infect Dis. 2003;56(27):107–9
  3. Munoz M, Liesenfeld O, Heimesaat MM. Immunology of Toxoplasma gondii. Immunol Rev. 2011;240(1):269–85
  4. Ganiem AR, Dian S, Indriati A, Chaidir L, Wisaksana R, Sturm P, et al. Cerebral Toxoplasmosis Mimicking Subacute Meningitis in HIV-Infected Patients; a Cohort Study from Indonesia. PLoS Negl Trop Dis. 2013;7(1):e1994
  5. Imran D, Estiasari R, Maharani K, Sucipto, Lestari DC, Yunus RE, et al. Presentation, etiology, and outcome of brain infections in an Indonesian hospital. Neurol Clin Pract. 2018 Oct 1;8(5):379 LP-388
  6. Robert-Gangneux F, Dardé ML. Epidemiology of and diagnostic strategies for toxoplasmosis. Clin Microbiol Rev. 2012;25(2):264–96
  7. Ferreira MS, Borges AS. Some Aspects of Protozoan Infections in Immunocompromised Patients: A Review. Mem Inst Oswaldo Cruz. 2002;97(4):443–57
  8. Meira CS, Vidal JE, Costa-Silva TA, Frazatti-Gallina N, Pereira-Chioccola VL. Immunodiagnosis in cerebrospinal fluid of cerebral toxoplasmosis and HIV-infected patients using Toxoplasma gondii excreted/secreted antigens. Diagn Microbiol Infect Dis. 2011;71(3):279–85
  9. Alfonso Y, Fraga J, Cox R, Bandera F, Pomier O, Fonseca C, et al. Comparison of four DNA extraction methods from cerebrospinal fluid for the detection of Toxoplasma gondii by polymerase chain reaction in AIDS patients. Med Sci Monit. 2008;14(3):MT1-6
  10. Lin MH, Chen TC, Kuo TT, Tseng CC, Tseng CP. Real-time PCR for quantitative detection of Toxoplasma gondii. J Clin Microbiol. 2000;38(11):4121–5
  11. Mesquita RT, Ziegler AP, Hiramoto RM, Vidal JE, Pereira-Chioccola VL. Real-time quantitative PCR in cerebral toxoplasmosis diagnosis of Brazilian human immunodeficiency virus-infected patients.J Med Microbiol. 2010;59(Pt 6):641–7
  12. Alfonso Y, Fraga J, Fonseca C, Jiménez N, Pinillos T, Dorta-Contreras AJ, et al. Molecular diagnosis of Toxoplasma gondii infection in cerebrospinal fluid from AIDS patients. Cerebrospinal Fluid Res. 2009;6:2
  13. Vidal JE, Colombo FA, Penalva AC, Oliveira D, Focaccia R, Lucia V, et al. PCR Assay Using Cerebrospinal Fluid for Diagnosis of Cerebral Toxoplasmosis in Brazilian AIDS patients PCR Assay Using Cerebrospinal Fluid for Diagnosis of Cerebral Toxoplasmosis in Brazilian AIDS patients. 2004;42(10):4765–8
  14. Alfonso Y, Fraga J, Cox R, Jiménez N, Capó V, Pomier O, et al. Conventional polymerase chain reaction for the diagnosis of neurotoxoplasmosis: comparison of three sets of primers for the B1 gene using CSF samples. Diagn Microbiol Infect Dis. 2013;75(2):150–4
  15. Derouin F, Leport C, Pueyo S, Morlat P, Letrillart B, Chene G, et al. Predictive value of Toxoplasma gondii antibody titres on the occurrence of toxoplasmic encephalitis in HIV-infected patients. ANRS 005/ACTG 154 Trial Group. AIDS. 1996 Nov;10(13):1521–7
  16. Hierl T, Reischl U, Lang P, Hebart H, Stark M, Kyme P, et al. Preliminary evaluation of one conventional nested and two real-time PCR assays for the detection of Toxoplasma gondii in immunocompromised patients. J Med Microbiol. 2004;53(7):629–32
  17. Teixeira LE, Kanunfre KA, Shimokawa PT, Targa LS, Rodrigues JC, Domingues W, et al. The performance of four molecular methods for the laboratory diagnosis of congenital toxoplasmosis in amniotic fluid samples. Rev Soc Bras Med Trop. 2013;46(5): 584-8
  18. Antioxidantitale M. A High Sensitive Nested PCR for Toxoplasma gondii Detection in Animal and Food Samples. J Microb Biochem Technol. 2013;05(02):39–41
  19. Jones CD, Okhravi N, Adamson P, Tasker S. 18S rDNA Genes of T . Gondii in Aqueous Humor AND. Invest Ophthalmol. 2000;41(3):634–44
  20. Grigg ME, Boothroyd JC. Rapid identification of virulent type I strains of the protozoan pathogen Toxoplasma gondii by PCR-restriction fragment length polymorphism analysis at the B1 gene. J Clin Microbiol. 2001;39(1):398–400
  21. Bastien P, Procop GW, Reischl U. Quantitative real-time PCR is not more sensitive than “ conventional” PCR. J Clin Microbiol. 2008;46(6):1897–900
  22. Paulo S. Toxoplasma- Specific IgG Subclass Antibody response in Cerebrospinal Fluid. 2015;57(5):439–42
  23. Ajzenberg D, Lamaury I, Demar M, Vautrin C, Cabié A, Simon S, et al. Performance Testing of PCR Assay in Blood Samples for the Diagnosis of Toxoplasmic Encephalitis in AIDS Patients from the French Departments of America and Genetic Diversity of Toxoplasma gondii : A Prospective and Multicentric Study. 2016;1–17
  24. Cingolani A, Luca ADE, Ammassari A, Murri R, Linzalone A, Grillo R, et al. PCR detection of Toxoplasma gondii DNA in CSF for the differential diagnosis of AIDS-related focal brain lesions. 2015;45(1996):7–11

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