Hashemizadeh Zahra, Bazargani Abdollah, Kalantar-Neyestanaki Davood, Mohebi Samane, Hadi Nahal
Department of Bacteriology and Virology, School of Medicine, Shiraz University of Medical Science, Shiraz, Iran.
Medical Mycology and Bacteriology Research Center, Kerman University of Medical Sciences, Kerman, Iran.
BMC Res Notes. 2020 Feb 24;13(1):97. doi: 10.1186/s13104-020-04948-z.
Molecular typing methods are useful for rapid detection and control of a disease. Recently, the use of high-resolution melting (HRM) for spa typing of MRSA isolates were reported. This technique is rapid, inexpensive and simple for genotyping and mutation screening in DNA sequence. The aim of this study was to evaluate the ability of HRM-PCR to analysis spa genes amongst MRSA isolates.
A total of 50 MRSA isolates were collected from two teaching hospitals in Shiraz, Iran. The isolates were confirmed as MRSA by susceptibility to cefoxitin and detection of mecA gene using PCR. We used HRM analysis and PCR-sequencing method for spa typing of MRSA isolates. In total, 15 different spa types were discriminate by HRM and sequencing method. The melting temperature of the 15 spa types, using HRM genotyping were between 82.16 and 85.66 °C. The rate of GC % content was 39.4-46.3. According to the results, spa typing of 50 clinical isolates via PCR-sequencing and HRM methods were 100% similar. Consequently, HRM method can easily identify and rapidly differentiate alleles of spa genes. This method is faster, less laborious and more suitable for high sample at lower cost and risk of contamination.
分子分型方法有助于疾病的快速检测与控制。最近，有报道称利用高分辨率熔解曲线分析（HRM）对耐甲氧西林金黄色葡萄球菌（MRSA）分离株进行spa分型。该技术在DNA序列的基因分型和突变筛查方面快速、廉价且操作简单。本研究旨在评估HRM-PCR分析MRSA分离株中spa基因的能力。
从伊朗设拉子的两家教学医院共收集了50株MRSA分离株。通过对头孢西丁的敏感性以及使用PCR检测mecA基因，确认这些分离株为MRSA。我们使用HRM分析和PCR测序方法对MRSA分离株进行spa分型。通过HRM和测序方法总共鉴别出15种不同的spa型。使用HRM基因分型时，这15种spa型的熔解温度在82.16至85.66°C之间。GC%含量为39.4 - 46.3。根据结果，通过PCR测序和HRM方法对50株临床分离株进行spa分型的结果100%相似。因此，HRM方法能够轻松识别并快速区分spa基因的等位基因。该方法更快、更省力，更适合处理大量样本，成本更低且污染风险更小。
