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基于蛋白A和凝固酶基因多态性的耐甲氧西林金黄色葡萄球菌临床分离株的分子分型

Molecular Typing of Methicillin Resistant Staphylococcus aureus Clinical Isolates on the Basis of Protein A and Coagulase Gene Polymorphisms.

作者信息

Omar Nancy Younis, Ali Hala Abdel Salam, Harfoush Reem Abdel Hameed, El Khayat Engy Hamdy

机构信息

Medical Microbiology & Immunology Department, Faculty of Medicine, Alexandria University, El Khartoom Square, El Azarita, Alexandria 21521, Egypt.

出版信息

Int J Microbiol. 2014;2014:650328. doi: 10.1155/2014/650328. Epub 2014 May 15.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a5a/4052513/7913e3ee4f3f/IJMICRO2014-650328.001.jpg

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