Esen Nuran, Ozkütük Aydan, Coban Hüseyin, Atlas Emek
Dokuz Eylül Üniversitesi Tıp Fakültesi, Tıbbi Mikrobiyoloji Anabilim Dalı, İzmir, Türkiye.
Mikrobiyol Bul. 2010 Jul;44(3):375-83.
Several methods are available for the molecular typing of Mycobacterium tuberculosis complex isolates. The results of the recent research demonstrated that Mycobacterial Interspersed Repetitive Unit (MIRU)-Variable Number Tandem Repeats (VNTR) method has high discriminatory power and reproducibility, is easy to perform, and available for multi-center studies and automation. However, there is insufficient data about the MIRU-VNTR profiles in Turkey. The aim of this study was to determine the most appropriate MIRU-VNTR combinations to distinguish cross contaminations and nosocomial infections in routine mycobacteriology laboratory practice. Following molecular typing of 152 clinical isolates which were consecutively isolated from different patients in two years period (August 2004-July 2006) in our laboratory, a retrospective analysis of MIRU-VNTR data of 12 loci primers was performed by an "in-house" computer based programme. The programme was prepared by using Microsoft QuickBASIC programming language and all of the data were calculated by the help of this programme. The best combinations to differentiate the clusters and to identify the unique isolates were determined out of 4095 possible results of 12 different primer pairs. According to our 152 MIRU-VNTR results, to determine cross contaminations and nosocomial infections in routine mycobacteriology laboratory practice, we recommend to use primers 26, 40, 16, 10 and 23 in the first step; primers 31, 27, 20 and 2 in the second step, and primers 4, 24 and 39 in the third step. The created software is user friendly, fast and meets the requirements of routine clinical mycobacteriology laboratories. Besides its discriminatory power, the speed and cost-effectiveness of a typing method is also considerable. According to the results of this study it was suggested that for more rapid and economic molecular typing of M.tuberculosis and related epidemiological investigations, MIRU-VNTR should be performed in a stepwise manner.
有几种方法可用于结核分枝杆菌复合群分离株的分子分型。近期研究结果表明，分枝杆菌散布重复单位（MIRU）-可变数目串联重复序列（VNTR）方法具有较高的鉴别力和可重复性，操作简便，可用于多中心研究和自动化。然而，土耳其关于MIRU-VNTR图谱的数据不足。本研究的目的是确定在常规分枝杆菌学实验室实践中区分交叉污染和医院感染的最合适的MIRU-VNTR组合。在对我们实验室在两年期间（2004年8月至2006年7月）从不同患者连续分离出的152株临床分离株进行分子分型后，通过一个基于“内部”计算机的程序对12个位点引物的MIRU-VNTR数据进行了回顾性分析。该程序使用Microsoft QuickBASIC编程语言编写，所有数据均借助此程序计算得出。从12种不同引物对的4095种可能结果中确定了区分聚类和鉴定独特分离株的最佳组合。根据我们的152个MIRU-VNTR结果，为了在常规分枝杆菌学实验室实践中确定交叉污染和医院感染，我们建议第一步使用引物26、40、16、10和23；第二步使用引物31、27、20和2；第三步使用引物4、24和39。所创建的软件用户友好、速度快，满足常规临床分枝杆菌学实验室的要求。除了其鉴别力外，分型方法的速度和成本效益也很重要。根据本研究结果，建议为了更快速、经济地对结核分枝杆菌进行分子分型及相关流行病学调查，应逐步进行MIRU-VNTR检测。
