Laboratoire de Génie Enzymatique et Microbiologie, Equipe Biotechnologie des Algues, Ecole Nationale d'Ingénieurs de Sfax, Université de Sfax, Sfax, Tunisia.
Laboratoire de Génie Enzymatique et Microbiologie, Equipe Biotechnologie des Algues, Ecole Nationale d'Ingénieurs de Sfax, Université de Sfax, Sfax, Tunisia.
Mol Cell Probes. 2020 Oct;53:101645. doi: 10.1016/j.mcp.2020.101645. Epub 2020 Jul 31.
Simkania negevensis is an emerging Chlamydia-like bacterium related to human respiratory diseases. An early and accurate detection of this pathogen could be useful to monitor the potential infectious risks and to set suitable outbreak control measures. In Tunisia, distribution and abundance of S. negevensis remain until now largely unknown. In the present work, a qPCR assay, targeting the 16S rRNA gene, for fast detection and quantification of S. negevensis was developed and validated. A high specificity for S. negevensis detection displaying no cross-reaction with the closely related Chlamydia spp. or the other tested microorganisms was noticed. qPCR assay performance was considered very satisfying with detection limits of 5 DNA copies per reaction. qPCR assay validation was performed by screening 37 clinical specimens and 35 water samples. S. negevensis wasn't detected in respiratory samples, but it was found in four cases of water samples. We suggest that the qPCR assay developed in this study could be considered sufficiently characterized to initiate the quantification of S. negevensis in environmental samples.
辛卡尼亚奈格威迪斯是一种新兴的衣原体样细菌，与人类呼吸道疾病有关。早期、准确地检测这种病原体可能有助于监测潜在的感染风险，并制定适当的暴发控制措施。在突尼斯，到目前为止，S. negevensis 的分布和丰度在很大程度上仍然未知。在本研究中，开发并验证了一种针对 16S rRNA 基因的 qPCR 检测方法，用于快速检测和定量 S. negevensis。该方法对 S. negevensis 的检测具有很高的特异性，与密切相关的衣原体属或其他测试的微生物没有交叉反应。qPCR 检测方法的性能被认为非常令人满意，检测限为每个反应 5 个 DNA 拷贝。qPCR 检测方法的验证是通过筛查 37 份临床标本和 35 份水样进行的。呼吸道样本中未检测到 S. negevensis，但在 4 份水样中发现了该细菌。我们建议，本研究中开发的 qPCR 检测方法可以被认为具有足够的特征，可以开始对环境样本中的 S. negevensis 进行定量分析。
