Omar Nancy Younis, Ali Hala Abdel Salam, Harfoush Reem Abdel Hameed, El Khayat Engy Hamdy
Medical Microbiology & Immunology Department, Faculty of Medicine, Alexandria University, El Khartoom Square, El Azarita, Alexandria 21521, Egypt.
Int J Microbiol. 2014;2014:650328. doi: 10.1155/2014/650328. Epub 2014 May 15.
Increased frequency of methicillin-resistant Staphylococcus aureus (MRSA) in hospitalized patients requires rapid and reliable characterization of isolates for control of MRSA spread in hospitals. This study evaluated polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) as a molecular typing technique for MRSA strains on the basis of protein A (spa) and coagulase (coa) gene polymorphisms to verify their ability in assessing the relatedness of isolates. Seventy-five MRSA isolates, from different ICUs of Alexandria University Main Hospital, were characterized using antibiotyping and PCR-RFLP analysis of coa and spa genes. Thirty-two antibiotypes were identified. coa gene PCR generated 3 types and 10 subtypes of band patterns. HaeIII restriction digestion of amplified coa gene products produced 5 major banding patterns and 12 subtypes. spa gene PCR products generated 4 major and 11 minor types, and their HaeII restriction digestion showed 5 major and 12 minor banding patterns. The combined coa and spa RFLP patterns generated 22 combined R types. Typing using coa PCR and PCR-RFLP had the same discriminatory index (DI) value (0.64), which was comparable to that of both spa PCR and PCR-RFLP techniques (0.68). The combined grouping increased the DI value to 0.836. The current study revealed that testing for multiple gene polymorphisms is more useful for local epidemiologic purposes.
住院患者中耐甲氧西林金黄色葡萄球菌(MRSA)感染频率的增加,要求快速且可靠地鉴定分离菌株,以控制MRSA在医院内的传播。本研究评估了聚合酶链反应-限制性片段长度多态性(PCR-RFLP)作为基于蛋白A(spa)和凝固酶(coa)基因多态性的MRSA菌株分子分型技术,以验证其评估分离菌株相关性的能力。使用亚历山大大学主医院不同重症监护病房的75株MRSA分离株,通过抗菌谱分型以及coa和spa基因的PCR-RFLP分析进行鉴定。共鉴定出32种抗菌谱型。coa基因PCR产生了3种类型和10种亚型的条带模式。扩增的coa基因产物经HaeIII限制性酶切产生5种主要条带模式和12种亚型。spa基因PCR产物产生4种主要类型和11种次要类型,其HaeII限制性酶切显示5种主要条带模式和12种次要条带模式。coa和spa RFLP联合模式产生了22种联合R型。使用coa PCR和PCR-RFLP分型具有相同的鉴别指数(DI)值(0.64),这与spa PCR和PCR-RFLP技术的DI值(0.68)相当。联合分组使DI值提高到0.836。当前研究表明,检测多个基因多态性对于局部流行病学研究更有用。