Mobasherizadeh Sina, Shojaei Hasan, Havaei Seyed Asghar, Mostafavizadeh Kamyar, Davoodabadi Fazollah, Khorvash Farzin, Ataei Behrooz, Daei-Naser Abbas
Nosocomial Infection Research Center, Isfahan University of Medical Sciences, Isfahan, Iran.
Glob J Health Sci. 2016 Aug 1;8(8):53822. doi: 10.5539/gjhs.v8n8p185.
The aim of this study was to apply RAPD technique to analyze the genetic variability among the Iranian CA-MRSA isolates.The RAPD amplification was implemented on 25 strains isolated from the anterior nares of 410 healthy children using four randomly selected oligonucleotide primers from the stocks available in our laboratory, including the primers 1254, GE6, OLP6 and OLP13 from our stock. The amplified PCR products were detected on a 1.5% agarose gel and subjected to further analysis to establish the band profiles and genetic relationships using the Gel Compar® program.The Iranian CA-MRSA isolates produced distinct RAPD patterns which varied based on the primer used, however, the primer 1254 revealed highly polymorphic patterns consisting 5 discernable RAPD types (RT), "RT1" (12, 48%), "RT2" (8, 32%), "RT3" (3, 12%), and "RT4 and RT5", (a single RAPD type each, 4%). Phylogenetic analysis based on RAPD profiles divided most of the CA-MRSA isolates into 2 distinct but related RAPD clusters, a small group and two single unrelated RAPD types.This study shows that the simple and cost-effective but rather difficult to optimize RAPD fingerprinting could be used to evaluate genetic and epidemiological relationships of CA-MRSA isolates on condition that the patterns are obtained from carefully optimized laboratory tests.
本研究的目的是应用随机扩增多态性DNA（RAPD）技术分析伊朗社区获得性耐甲氧西林金黄色葡萄球菌（CA-MRSA）分离株之间的遗传变异性。使用从我们实验室现有的引物库中随机选择的4种寡核苷酸引物，对从410名健康儿童的前鼻孔分离出的25株菌株进行RAPD扩增，包括我们引物库中的引物1254、GE6、OLP6和OLP13。扩增的PCR产物在1.5%的琼脂糖凝胶上进行检测，并使用Gel Compar®程序进行进一步分析，以建立条带图谱和遗传关系。伊朗CA-MRSA分离株产生了不同的RAPD模式，这些模式因所用引物而异，然而，引物1254显示出高度多态性的模式，由5种可识别的RAPD类型（RT）组成，“RT1”（12株，48%），“RT2”（8株，32%），“RT3”（3株，12%），以及“RT4和RT5”（各为单一RAPD类型，4%）。基于RAPD图谱的系统发育分析将大多数CA-MRSA分离株分为2个不同但相关的RAPD簇、一个小群体和两种单一的不相关RAPD类型。本研究表明，简单且经济高效但难以优化的RAPD指纹图谱可用于评估CA-MRSA分离株的遗传和流行病学关系，前提是这些模式是通过精心优化的实验室测试获得的。
