Development of a new TaqMan-based PCR assay for the specific detection and quantification of Simkania negevensis. (October 2020)

• Record Type: Journal Article
• Authors: Baccari, Olfa; Elleuch, Jihen; Barkallah, Mohamed; Boukedi, Hanen; Ayed, Nourelhouda Ben; Hammami, Adnene; Fendri, Imen; Abdelkafi, Slim
• Is Part Of: Molecular and cellular probes. Volume 53(2020)
• Publication Date: 2020-10
• Subjects: Simkania negevensis -- Detection -- qPCR assay -- TaqMan -- Water -- Tunisia; Molecular probes -- Diagnostic use -- Periodicals; Pathology, Cellular -- Technique -- Periodicals; Cell Biology -- Periodicals; Molecular Biology -- Periodicals; Sondes moléculaires -- Utilisation diagnostique -- Périodiques; Cytopathologie -- Technique -- Périodiques; 572
• DOI: 10.1016/j.mcp.2020.101645
• Languages: English
• ISSNs: 0890-8508

Abstract

Abstract: 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. Highlights: A TaqMan qPCR assay was designed for both the detection and the absolute quantification of an emerging Chlamydia -like bacterium called Simkania negevensis. 100% specificity and sensitivity. A low detection limit at 5 DNA copies per reaction as well as high reproducibility with a coefficient of variation value ranged from 0.2 to 2.59%. Successful applications of designed qPCR assay for S . negevensis detection and quantification in environmental samples.