耐甲氧西林金黄色葡萄球菌两种基因分型方法的对比研究  

Comparative observation on the two methods for genotyping methicillin-resistant Staphylococcus aureus

作　　者：莫晓能[1] 李建国[1] 唐英春[2] 张永[2] 朱家馨[2] 谈淑卿[2] 陆坚[2] 

机构地区：[1]中山大学第二附属医院呼吸内科,广东广州510120 [2]中山大学第三附属医院,广东广州510060

出　　处：《中国热带医学》2006年第8期1333-1335,1349,共4页China Tropical Medicine

基　　金：广东省卫生厅课题资助(课题编号A2004552

摘　　要：目的比较耐甲氧西林金黄色葡萄球菌(MRSA)两种基因分型方法的特点,为MRSA流行病学调查选用合适的基因分型方法提供参考。方法用聚合酶链反应(PCR)扩增MRSA基因高变区(Hypervariable region,HVR),并根据扩增片段大小进行基因分型,同时与ERIC-2随机引物多态性DNA分型法进行比较,并观察不同退火温度条件下两种基因分型方法的差异。结果50株MRSA以基因高变区(HVR)分型,可分为A、B、C、D E 5种基因型,其中以B(47.3%)和D(32.1%)型多见,A(3.5%)、C(10.7%)、E(6.4%)型少见,当退火温度改变时,则不能分型;用ERIC-2随机引物对MRSA进行多态性DNA分型,在低退火温度下分为10种基因型,当退火温度明显改变时,仍能分型,但每型的扩增片断明显减少。结论随机引物多态性DNA分型和HVR基因分型两种方法均能用于MRSA感染的流行病学研究,HVR基因分型PCR反应温度控制严格,分型明确;而随机引物多态性ERIC-2的DNA分型随PCR退火温度的变化而变化,应用时需严格控制PCR退火温度等反应参数,否则重复性较差。Objective To study the characteristic of two methods used in genotyping of methicillin resistant Staphylococcus aureus （MRSA） and provide reference for carrying out investigation on molecular epidemiology. Methods A total of 50 non - duplicate MRSA clinical isolates were collected in 4 hospitals of Guangdong Province. Special primer associated with hyper variable region （HVR） and REIC - 2 were used for amplification of MRSA BY PCR and the genotypes were differentiated based on the fragments of amplified products. The difference of two genotypes was observed according to the change of the fragments of amplified products when the annealing temperature was changed. Results 50 strains of MRSA were defferentiated into the genotpyes of A, B, C, D and E based on HVR genotypes and into 10 genotypes on ERIC - 2 pelymorphism; Moreover, there were still genotypes by ERIC - 2 primer when annealing temperature of PCR was changed to 64℃. Conclusion Either Randomly amplified pelymorphic DNA analysis（RAPD） ERIC - 2 or genotype of HVR（ HVR - PCR） is an rapid, reliable and effective method for epidemiological study on MRSA at molecular level. But the genotying results will alter along with the changes of annealing temperature. Therefore, the annealing temperature should be controlled strictly when RAPD is used, otherwise, the results of genotype may be not reliable.

关 键 词：耐甲氧西林金黄色葡萄球菌 基因分型 对比研究 

分 类 号：R378.11[医药卫生—病原生物学]