Yun Kyung Won, Song Eun Ju, Choi Go Eun, Hwang In Kyung, Lee Eun Yup, Chang Chulhun L
Department of Internal Medicine, Busan St. Mary's Hospital, Busan, Korea.
Korean J Lab Med. 2009 Aug;29(4):314-9. doi: 10.3343/kjlm.2009.29.4.314.
Mycobacterium tuberculosis is one of the most clinically significant infectious agents. Especially during mass outbreaks, accurate identification and monitoring are required. The proportion of Beijing family members is very high among infecting strains, and spoligotyping is not suitable for strain typing. Therefore, we studied the homogeneity of isolates using the mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) method and identified its utility for carrying out molecular epidemiologic analysis.
Eighty-one clinical M. tuberculosis isolates that had previously been analyzed by spoligotyping were used in this study. We used the 12 standard MIRU loci and further four exact tandem repeat (ETR) loci (ETR-A, -B, -C, and -F). Four strains each of randomly selected Beijing and Beijing-like families were subjected to IS6110- restriction fragment length polymorphism analysis.
All 81 samples showed amplification products of all VNTR loci, and all of them showed differences in at least one locus. The calculation of the Hunter-Gaston diversity index (HGDI) for MIRU-VNTR gave the value of 0.965. Discriminatory index in the six loci (MIRU-10, -16, -26, -31, -39, and ETR-F) were found to be highly discriminated (HGDI >0.6). Beijing and Beijing-like family isolates were discriminated into different MIRU-VNTR types.
MIRU-VNTR analysis by using well-selected loci can be useful in discriminating the clinical M. tuberculosis isolates in areas where the Beijing family is predominant.
结核分枝杆菌是临床上最重要的感染病原体之一。尤其是在大规模疫情爆发期间,需要进行准确的鉴定和监测。在感染菌株中,北京家族成员的比例非常高,而间隔寡核苷酸分型法(spoligotyping)不适用于菌株分型。因此,我们使用分枝杆菌散布重复单位-可变数目串联重复序列(MIRU-VNTR)方法研究了分离株的同质性,并确定了其在进行分子流行病学分析中的实用性。
本研究使用了81株先前已通过间隔寡核苷酸分型法分析的临床结核分枝杆菌分离株。我们使用了12个标准MIRU位点以及另外4个精确串联重复序列(ETR)位点(ETR-A、-B、-C和-F)。对随机选择的4株北京家族和类北京家族菌株分别进行IS6110-限制性片段长度多态性分析。
所有81个样本均显示所有VNTR位点的扩增产物,且所有样本在至少一个位点上存在差异。MIRU-VNTR的Hunter-Gaston多样性指数(HGDI)计算值为0.965。发现6个位点(MIRU-10, -16, -26, -31, -39和ETR-F)的鉴别指数具有高度鉴别力(HGDI>0.6)。北京家族和类北京家族分离株被鉴别为不同的MIRU-VNTR类型。
通过使用精心选择的位点进行MIRU-VNTR分析,可有助于鉴别北京家族占主导地位地区的临床结核分枝杆菌分离株。