Aitlhaj-Mhand Rokaya, Qasmaoui Aicha, Bellaji Bahija, Remz Chaimae, Charof Reda, Jaoudi Rachid El, Abdelmoumen Hanaa, Hançali Amina, Oumzil Hicham
Virology Department, National Institute of Hygiene, Rabat, Morocco.
Microbiology and Molecular Biology Team, Research Center for Plant and Microbial Biotechnology, Biodiversity and Environment, Faculty of Sciences, Mohammed V University in Rabat, Rabat, Morocco.
Infez Med. 2024 Sep 1;32(3):352-362. doi: 10.53854/liim-3203-9. eCollection 2024.
Gonorrhea is a prevalent sexually transmitted infection among men who have sex with men (MSM). In Morocco, the basic laboratory diagnosis of (NG) is based on microscopy and, in some settings, on culture. However, no nucleic acid amplification test (NAAT) has been implemented for routine diagnosis of gonorrhoeae.The aim of this study is to assess the effectiveness of an in-house real-time PCR test for detecting DNA in anal swabs samples collected during an Integrated Behavioral and Biological survey.
Samples from 245 MSM, recruited using a Respondent Driven Sampling, were collected and tested for NG infection using GeneXpert CT/NG assay (Cepheid, USA). An In-House real-time PCR technique targeting the pseudo gene porA was developed and used for a parallel investigation of the same infection. The reliability of the in-house RT-PCR was validated through tests of reproducibility, repeatability, limit of detection, and cross-reactivity with other bacteria. The intrinsic performance characteristics of the qRT-PCR were assessed, namely, the sensitivity, the specificity, the positive predictive value (PPV), and the negative predictive value (NPV). The GeneXpert CT/NG assay was adopted as a reference method.
For detection, the in-house real-time PCR assay showed a sensitivity and specificity of 80% and 100%, respectively. The PPV of the assay was 100% and the NPV was 97.3%.
The in-house real-time PCR assay has high specificity and sensitivity, and it emerges as a promising approach for detecting in clinical specimens, particularly in decentralized settings such as regional laboratories.
淋病是男男性行为者（MSM）中一种常见的性传播感染。在摩洛哥，淋病奈瑟菌（NG）的基本实验室诊断基于显微镜检查，在某些情况下还基于培养。然而，尚未实施核酸扩增试验（NAAT）用于淋病的常规诊断。本研究的目的是评估一种内部实时聚合酶链反应（PCR）检测方法在综合行为和生物学调查期间收集的肛门拭子样本中检测淋病奈瑟菌DNA的有效性。
使用应答驱动抽样招募了245名男男性行为者的样本，并使用GeneXpert CT/NG检测法（美国赛沛公司）对其进行淋病感染检测。开发了一种针对porA假基因的内部实时PCR技术，并用于对同一感染进行平行调查。通过重复性、再现性、检测限以及与其他细菌的交叉反应性测试，验证了内部实时PCR的可靠性。评估了定量逆转录PCR（qRT-PCR）的内在性能特征，即敏感性、特异性、阳性预测值（PPV）和阴性预测值（NPV）。采用GeneXpert CT/NG检测法作为参考方法。
对于淋病奈瑟菌检测，内部实时PCR检测法的敏感性和特异性分别为80%和100%。该检测法的PPV为100%，NPV为97.3%。
内部实时PCR检测法具有高特异性和敏感性，是一种在临床标本中检测淋病奈瑟菌的有前景的方法，特别是在区域实验室等分散的环境中。
