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. 2019 Mar;33(3):e22719.
doi: 10.1002/jcla.22719. Epub 2018 Nov 25.

Combined testing for herpes simplex virus and Mycobacterium tuberculosis DNA in cerebrospinal fluid of patients with aseptic meningitis in Burkina Faso, West Africa

Sylvie Zida  1   2 , Pratt Kolia-Diafouka  1 , Dramane Kania  2 , Albert Sotto  3 , Vincent Foulongne  1 , Karine Bolloré  1 , Soumeya Ouangraoua  2 , Nicolas Méda  4 , Séverine Carrère-Kremer  1 , Philippe Van de Perre  1 , Edouard Tuaillon  1
Affiliations

Combined testing for herpes simplex virus and Mycobacterium tuberculosis DNA in cerebrospinal fluid of patients with aseptic meningitis in Burkina Faso, West Africa

Sylvie Zida et al. J Clin Lab Anal. 2019 Mar.

Abstract

Background: Little is known about the involvement of herpes simplex virus (HSV) or Mycobacterium tuberculosis (MTB) as potentially curable causes of central nervous system (CNS) infections in sub-Saharan Africa.

Objective: In this study, we developed a PCR assay dedicated to simultaneous testing of HSV1/HSV2 and MTB in Burkina Faso, a country where HSV is neglected as a cause of CNS infection and where TB prevalence is high.

Methods: A consensus HSV1/HSV2 set of primers and probe were designed and combined to primers and probe targeting the IS6110 repetitive insertion sequence of MTB. Analytical performances of the assay were evaluated on reference materials. Cerebrospinal fluid (CSF) collected from subjects with aseptic meningitis was tested for HSV1/HSV2 and MTB DNA.

Results: The UL29 gene was chosen as a highly conserved region targeted by the HSV1/HSV2 nucleic acid test. The lower limits of detection were estimated to be 2.45 copies/µL for HSV1, 1.72 copies/µL for HSV2, and 2.54 IS6110 copies per µL for MTB. The PCR was used in 202 CSF collected from subjects suspected of aseptic meningitis. Five samples (2.46%) tested positive, including two children positive for HSV1 (0.99%) and three adults tested positive for MTB (1.47%).

Conclusion: Using an in-house real-time PCR assay, we showed that both HSV and MTB are etiologic pathogens contributing to aseptic meningitis in Burkina Faso. This molecular test may have clinical utility for early diagnosis for those treatable CNS infections.

Keywords: Mycobacterium tuberculosis; PCR; cerebrospinal fluid; herpes simplex virus; meningitis.

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Figures

Figure 1
Figure 1
Detection limits of the HSV1/HSV2, and IS61160 PCR assays. Curves were determined by probit analysis (95% probability detection). A, The LOD was estimated at 2.45 copies/µL for HSV1; B, 1.72 copies/µL for HSV2; and C, 2.54 copies/µL for IS6110 using the BCG strain containing a single target copy of IS6110. Dashed lines show 95% confidence interval for the analysis. BCG, bacillus Calmette‐Guerin; HSV, herpes simplex virus; LOD, limits of detection
Figure 2
Figure 2
Duplex HSV1/HSV2 and Mycobacterium tuberculosis PCR standard curves established by means of serial dilutions of DNA. The HSV1 standard curve is represented by (A), the HSV2 standard curve by (B), and the BCG DNA standard curve by (C). BCG, bacillus Calmette‐Guerin; HSV, herpes simplex virus
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