用于临床分离株中基因、和快速检测的高分辨率熔解曲线分析的发展。

Development of high-resolution melting curve analysis in rapid detection of gene, , and from clinical isolates.

作者信息

Dehbashi Sanaz, Tahmasebi Hamed, Sedighi Parinaz, Davarian Faeze, Arabestani Mohammad Reza

机构信息

1Department of Microbiology, School of Medicine, Hamadan University of Medical Sciences, Pajoohesh junction, Hamadan, Iran.

2Department of Microbiology, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran.

出版信息

Trop Med Health. 2020 Feb 18;48:8. doi: 10.1186/s41182-020-00197-9. eCollection 2020.

BACKGROUND

High-resolution melting analysis (HRMA) is a novel molecular technique based on the real-time PCR that can be used to detect vancomycin resistance Enterococcus (VRE). The purpose of this study was to identify VRE species with HRMA in clinical isolates.

RESULTS

Out of 49 Enterococcus isolates, 11 (22.44%) isolates and 19 (38.77%) isolates were detected. Average melting temperatures for in , in , and in VRE strains were obtained as 79.9 ± 0.5 °C, 85.4 ± 0.5 °C, and 82.99 ± 0.5 °C, respectively. Furthermore, the data showed that the HRMA method was sensitive to detect 10 CFU/ml for the , , and genes. Also, out of 49 Enterococcus spp., which were isolated by HRMA assay, 8 isolates (16.32%) of and 18 isolates (36.73%) of were detected. The gene was reported in 2 isolates (25%) of and 9 isolates (50%) of .

CONCLUSIONS

This study demonstrated that using the HRMA method, we can detect , , and the gene with high sensitivity and specificity.

背景

高分辨率熔解分析（HRMA）是一种基于实时聚合酶链反应的新型分子技术，可用于检测耐万古霉素肠球菌（VRE）。本研究的目的是用HRMA鉴定临床分离株中的VRE种类。

结果

在49株肠球菌分离株中，检测到11株（22.44%）和19株（38.77%）分离株。粪肠球菌、屎肠球菌和鹑鸡肠球菌VRE菌株的平均熔解温度分别为79.9±0.5°C、85.4±0.5°C和82.99±0.5°C。此外，数据表明HRMA方法对检测粪肠球菌、屎肠球菌和鹑鸡肠球菌基因的灵敏度为10 CFU/ml。而且，在通过HRMA检测分离出的49株肠球菌中，检测到8株（16.32%）粪肠球菌和18株（36.73%）屎肠球菌。在2株（25%）粪肠球菌和9株（50%）屎肠球菌中检测到vanA基因。