Barmak Sabyrkhan M, Sinyavskiy Yuriy A, Berdygaliev Aidar B, Sharmanov Turegeldy Sh, Savitskaya Irina S, Sultankulova Gulmira T, Zholdybayeva Elena V
Faculty of Biology and Biotechnology, Al-Farabi Kazakh National University, Almaty 050040, Kazakhstan.
Biotechnology and Biologically Active Substances Laboratory, Kazakh Academy of Nutrition, Almaty 050008, Kazakhstan.
Microorganisms. 2021 Nov 9;9(11):2319. doi: 10.3390/microorganisms9112319.
In this study, we aimed to compare the performance of conventional PCR and real-time PCR assays as screening methods for identification of three frequent, clinically significant Salmonella serovars in Kazakhstan. We determined the diagnostic efficacy of three molecular methods for detection of subsp. enterica and typing Typhimurium, Enteritidis, and Virchow. A total of 137 clinical samples and 883 food samples were obtained in Almaty in 2018-2019. All tests showed high analytical specificity for detecting and its corresponding serovariants (100%). The sensitivity of real-time PCR for each of the tested targets was 1-10 microbial cells and in conventional PCR 10-100 microbial cells. The trials with conventional PCR and real-time PCR had a diagnostic efficacy (DE) of 100% and 99.71%, respectively. The DE of real-time PCR and conventional PCR for detecting Enteritidis and Typhimurium was 99.90%, while the DE of conventional PCR and real-time PCR for detecting Virchow was 99.31% and 99.80%, respectively. The RAPD-PCR analysis of the genomic DNA of showed the genetic kinship of Enteritidis isolates, and the genetic heterogeneity of Typhimurium and Virchow isolates. Thus, the developed methods can be considered as alternatives to classical serotyping using antisera.
在本研究中，我们旨在比较常规PCR和实时PCR检测方法在哈萨克斯坦作为三种常见且具有临床意义的沙门氏菌血清型鉴定筛选方法的性能。我们确定了三种分子方法检测肠炎沙门氏菌亚种及对鼠伤寒沙门氏菌、肠炎沙门氏菌和维尔肖沙门氏菌进行分型的诊断效能。2018 - 2019年在阿拉木图共采集了137份临床样本和883份食品样本。所有检测对检测肠炎沙门氏菌及其相应血清型均显示出高分析特异性（100%）。实时PCR对每个检测靶点的灵敏度为1 - 10个微生物细胞，常规PCR为10 - 100个微生物细胞。常规PCR和实时PCR试验的诊断效能（DE）分别为100%和99.71%。实时PCR和常规PCR检测肠炎沙门氏菌和鼠伤寒沙门氏菌的DE为99.90%，而常规PCR和实时PCR检测维尔肖沙门氏菌的DE分别为99.31%和99.80%。对肠炎沙门氏菌基因组DNA的RAPD - PCR分析显示了肠炎沙门氏菌分离株的遗传亲缘关系，以及鼠伤寒沙门氏菌和维尔肖沙门氏菌分离株的遗传异质性。因此，所开发的方法可被视为使用抗血清进行经典血清分型的替代方法。
