Zida Sylvie, Kolia-Diafouka Pratt, Kania Dramane, Sotto Albert, Foulongne Vincent, Bolloré Karine, Ouangraoua Soumeya, Méda Nicolas, Carrère-Kremer Séverine, Van de Perre Philippe, Tuaillon Edouard
UMR 1058, INSERM/EFS/Université de Montpellier, Montpellier, France.
Centre MURAZ, Bobo Dioulasso, Burkina Faso.
J Clin Lab Anal. 2019 Mar;33(3):e22719. doi: 10.1002/jcla.22719. Epub 2018 Nov 25.
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.
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.
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.
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%).
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.
在撒哈拉以南非洲地区，对于单纯疱疹病毒（HSV）或结核分枝杆菌（MTB）作为中枢神经系统（CNS）感染潜在可治愈病因的参与情况知之甚少。
在本研究中，我们开发了一种PCR检测方法，专门用于在布基纳法索同时检测HSV1/HSV2和MTB。在该国，HSV作为CNS感染病因被忽视，且结核病患病率很高。
设计了一组针对HSV1/HSV2的共识引物和探针，并与靶向MTB的IS6110重复插入序列的引物和探针相结合。在参考材料上评估该检测方法的分析性能。对从无菌性脑膜炎患者收集的脑脊液（CSF）进行HSV1/HSV2和MTB DNA检测。
选择UL29基因作为HSV1/HSV2核酸检测的高度保守区域。HSV1的检测下限估计为2.45拷贝/微升，HSV2为1.72拷贝/微升，MTB的IS6110拷贝数下限为每微升2.54拷贝。PCR用于检测从疑似无菌性脑膜炎患者收集的202份CSF。5份样本（2.46%）检测呈阳性，包括2名HSV1阳性儿童（0.99%）和3名MTB阳性成人（1.47%）。
通过使用内部实时PCR检测方法，我们表明HSV和MTB都是导致布基纳法索无菌性脑膜炎的病原病原体。这种分子检测方法对于那些可治疗的CNS感染的早期诊断可能具有临床实用性。
