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作 者:梁佳元[1] 孙蕾[1] 王毳[1] LIANG Jiayuan;SUN Lei;WANG Cui(Liaoning Center for Disease Prevention and Control, Shenyang, Liaoning 110005, China)
机构地区:[1]辽宁省疾病预防控制中心,辽宁沈阳110005
出 处:《中国热带医学》2018年第6期528-530,共3页China Tropical Medicine
基 金:辽宁省自然科学基金(No.2015020562)
摘 要:目的为明确比较RD105基因缺失法和双重PCR法对辽宁地区结核分枝杆菌中北京家族菌株的检测能力,同时掌握辽宁地区结核杆菌中北京家族菌株的流行情况而进行该项研究。方法选取2011—2013年间辽宁省内各地市临床分离菌株,随机抽取经传统方法菌种鉴定为结核分枝杆菌的菌株,共计125株。选用超声破碎法提取菌株DNA,应用RD105基因缺失法和双重PCR法,同时对收集到的125株结核分枝杆菌进行特异性扩增,最后使用琼脂糖凝胶水平电泳对扩增的结果进行判断。结果两种方法判断标准为:RD105基因缺失法结果中,出现283 bp大小片段的为北京家族菌株,不出现特异性条带的为非北京家族菌株;双重PCR法结果中,只出现393 bp大小片段的为北京家族菌株,而只出现570 bp大小片段为非北京家族菌株。使用上述两种方法鉴定本研究的125株结核分枝杆菌,结果完全相同,其中北京家族菌株有110株(88%),非北京家族菌株有15株(12%)。结论 RD105基因缺失法和双重PCR法对辽宁地区结核分枝杆菌中北京家族菌株的检测能力相当,并且北京家族菌株在辽宁地区流行的结核分枝杆菌中为绝对优势株;两种方法相对于传统的Spoligotyping法更简单、更省时,检测成本也较低,更适合在我国多数实验室推广应用。Objective To compare the detection ability of RD105 deletion test and duplex PCR for “Beijing family” strain of Mycobacterium tuberculosis (MTB), and to master the epidemic situation of “Beijing family” strains in Liaoning Province. Methods A total of 125 strains identified as MTB were randomly selected as clinical isolates from different cities in Liaoning Province between 2011 and 2013. MTB DNA of these strains were extracted by sonication. The 125 strains of MTB were specific amplified by RD105 deletion test and duplex PCR. Finally, the amplification results were judged by agarose gel electrophoresis. Results Using the RD105 deletion test, the 283 bp size fragment was a Beijing family strain and the negative result of electrophoresis was non-Beijing family strain; Using the duplex PCR, only 393 bp size fragment was Beijing family strain and only 570 bp size fragment was non-Beijing family strain. The identifying results of 125 MTB of the two methods were the same, 110 were determined as “Beijing family” strain (88%), and 15 were determined as “non-Beijing family” strain (12%). Conclusions Both methods have the same detection capability for “Beijing family” strains, and “Beijing family” strains predominate in epidemic strains in Liaoning Province. Compared with the traditional Spoligotyping method, the two methods are simpler, more time-saving, and the detection cost is lower, which is more suitable for popularization and application in most laboratories in China.
关 键 词:结核分枝杆菌 北京家族 RD105基因缺失法 双重PCR法
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