CAJ | 학술논문

目的初步评价不同串联重复序列(VNTR)位点在中国8省市结核分枝杆菌基因分型中的应用,寻找适合中国地区结核分枝杆菌基因分型的位点组合.方法从中国8个省(市、自治区)2800余株结核分枝杆菌临床分离菌株中以简单数字表法随机抽取140株,采用多位点数目可变串联重复序列分析方法(MLVA)对27个数目可变VNTR位点进行基因多态性检测,采用BioNumerics数据库软件进行单位点和不同位点组合的分辨率(Hunter-Gaston指数,HGI)分析,并比较分析其对140株菌的基因分型鉴定能力.同时采用间隔区寡核苷酸分型(Spoligotyping)将140株菌分为北京家族和非北京家族,评价上述不同VNTR位点组合在北京家族和非北京家族中的分型能力.结果 140株菌主要可分为2个基因群,即北京家族112株,占80%;非北京家族28株,占20%.Spoligotyping分型对140株结核分枝杆菌的HGI为0.4589.MLVA分析结果显示不同位点在不同菌株群存在明显的多态性,不同位点的HGI具有较大差异(0～0.809),对全部菌株、北京家族菌株、非北京家族菌株的HGI达到0.5以上的VNTR位点数分别为8、7和14个.27个VNTR位点进行不同的位点组合:优化筛选的8位点组合、国际推荐的12个、15个和24个位点组合.4个组合的HGl分别为0.9991、0.9882、0.9980和0.9986;在北京家族菌株中,上述组合的HGI依次为0.9987、0.9318、0.9969和0.9975;在非北京家族菌株中分别为1、0.9894、1和1.结论不同的VNTR位点和不同VNTR位点组合在中国8省市结核分枝杆菌中的HGI均存在明显差异;本研究优化的8个位点组合MLVA分型方法在中国结核分枝杆菌流行病学研究可能具有良好的应用前景.
목적 초보평개불동천련중복서렬(VNTR)위점재중국8성시결핵분지간균기인분형중적응용,심조괄합중국지구결핵분지간균기인분형적위점조합.방법 종중국8개성(시、자치구)2800여주결핵분지간균림상분리균주중이간단수자표법수궤추취140주,채용다위점수목가변천련중복서렬분석방법(MLVA)대27개수목가변VNTR위점진행기인다태성검측,채용BioNumerics수거고연건진행단위점화불동위점조합적분변솔(Hunter-Gaston지수,HGI)분석,병비교분석기대140주균적기인분형감정능력.동시채용간격구과핵감산분형(Spoligotyping)장140주균분위북경가족화비북경가족,평개상술불동VNTR위점조합재북경가족화비북경가족중적분형능력.결과 140주균주요가분위2개기인군,즉북경가족112주,점80%;비북경가족28주,점20%.Spoligotyping분형대140주결핵분지간균적HGI위0.4589.MLVA분석결과현시불동위점재불동균주군존재명현적다태성,불동위점적HGI구유교대차이(0～0.809),대전부균주、북경가족균주、비북경가족균주적HGI체도0.5이상적VNTR위점수분별위8、7화14개.27개VNTR위점진행불동적위점조합:우화사선적8위점조합、국제추천적12개、15개화24개위점조합.4개조합적HGl분별위0.9991、0.9882、0.9980화0.9986;재북경가족균주중,상술조합적HGI의차위0.9987、0.9318、0.9969화0.9975;재비북경가족균주중분별위1、0.9894、1화1.결론불동적VNTR위점화불동VNTR위점조합재중국8성시결핵분지간균중적HGI균존재명현차이;본연구우화적8개위점조합MLVA분형방법재중국결핵분지간균류행병학연구가능구유량호적응용전경.
Objective To evaluate the application of different variable number tandem repeats (VNTR) locus in genotyping of Mycobacterium tuberculosis (M. tuberculosis) strains isolated from eight provinces in China, and to find the suitable locus-set of VNTR for epidemical strains in China. Methods All 140 M. tuberculosis strains were randomly selected from 2800 M. tuberculosis strains isolated from eight provinces in China,27 VNTR loci were used for typing all isolates. Discriminatory power (Hunter-Gaston Index, HGI) of every locus and different locus-set were analyzed by BioNumerics software. Meanwhile, Spoligotyping was used to identify Beijing family and non-Beijing family. Then the HGI of different locus-sets in two families was also evaluated. Results All 140 isolates were clustered into Beijing kindred (112 strains,80%) and non-Beijing kindred (28 strains, 20%) by Spoligotyping. The discriminatory power of Spoligotyping in 140 isolates was 0.4589. Every locus showed different polymorphism and HGI were from 0 to 0.809. The number of VNTR loci with HGI higher than 0.5 in all strains, Beijing family and non-Beijing family was 8,7 and 14 respectively. 27 loci were combined into four groups which included 8,12,15 and 24 VNTR loci respectively. Four locus-sets showed different polymorphism, HGI of eight-locus, 12-locus, 15-locus,and 24-locus set in 140 strains was 0.9991,0.9882,0.9980 and 0.9986, and their discriminatory power were calculated in Beijing kindred (HGI: 0.9987, 0.9318, 0.9969 and 0.9975) and non-Beijing kindred (HGI: 1,0.9894,1 and 1). Conclusion Different VNTR locus and locus-set showed different discriminatory power in the selected M. tuberculosis strains isolated from China. Eight-locus set can be used in molecular epidemiological study of M. tuberculosis in China after standardization.