Werner Guido, Klare Ingo, Witte Wolfgang
Robert Koch Institute, Wernigerode Branch, Wernigerode, Germany.
BMC Microbiol. 2007 Apr 10;7:28. doi: 10.1186/1471-2180-7-28.
MLVA (multiple-locus variable-number tandem repeat analysis) is a reliable typing technique introduced recently to differentiate also isolates of Enterococcus faecium. We used the established VNTR (variable number of tandem repeats) scheme to test its suitability to differentiate 58 E. faecium isolates representing mainly outbreaks and clusters of infections and colonizations among patients from 31 German hospitals. All isolates were vancomycin-resistant (vanA type). Typing results for MLVA are compared with results of macrorestriction analysis in PFGE (pulsed-field gel electrophoresis) and MLST (multi-locus sequence typing).
All 51 but one hospital isolates from 1996-2006 were assigned to the clonal complex (CC) of epidemic-virulent, hospital-adapted lineages (MLST CC-17; MLVA CC-1) and differed from isolates of sporadic infections and colonizations (n = 7; 1991-1995) and other non-hospital origins (n = 27). Typing of all 58 hospital VRE revealed MLVA as the least discriminatory method (Simpson's diversity index 0.847) when compared to MLST (0.911) and PFGE (0.976). The two most common MLVA types MT-1 (n = 16) and MT-159 (n = 14) combined isolates of several MLST types including also major epidemic, hospital-adapted, clonal types (MT-1: ST-17, ST-18, ST-280, ST-282; MT-159: ST-78, ST-192, ST-203). These data clearly indicate that non-related E. faecium could possess an identical MLVA type being especially critical when MLVA is used to elucidate supposed outbreaks with E. faecium within a single or among different hospitals. Stability of a given MLVA profile MT-12 (ST-117) during an outbreak over a period of five years was also shown.
MLVA is a suitable method to assign isolates of E. faecium into distinct clonal complexes. To investigate outbreaks the current MLVA typing scheme for E. faecium does not discriminate enough and cannot be recommended as a standard superior to PFGE.
多位点可变数目串联重复序列分析(MLVA)是一种最近引入的可靠分型技术,也用于区分粪肠球菌分离株。我们使用已建立的可变数目串联重复序列(VNTR)方案来测试其区分58株粪肠球菌分离株的适用性,这些分离株主要代表来自31家德国医院患者中的感染和定植暴发及聚集情况。所有分离株均对万古霉素耐药(vanA型)。将MLVA的分型结果与脉冲场凝胶电泳(PFGE)中的宏观限制性分析结果和多位点序列分型(MLST)结果进行比较。
1996 - 2006年来自所有51家医院(仅1家除外)的分离株被归入流行 - 毒力、医院适应性谱系的克隆复合体(CC)(MLST CC - 17;MLVA CC - 1),与散发性感染和定植的分离株(n = 7;1991 - 1995年)以及其他非医院来源的分离株(n = 27)不同。对所有58株医院耐万古霉素肠球菌进行分型发现,与MLST(0.911)和PFGE(0.976)相比,MLVA是区分能力最弱的方法(辛普森多样性指数为0.847)。两种最常见的MLVA类型MT - 1(n = 16)和MT - 159(n = 14)合并了几种MLST类型的分离株,其中还包括主要的流行、医院适应性克隆类型(MT - 1:ST - 17、ST - 18、ST - 280、ST - 282;MT - 159:ST - 78、ST - 192、ST - 203)。这些数据清楚地表明,不相关的粪肠球菌可能具有相同的MLVA类型,这在使用MLVA阐明单个医院或不同医院内疑似粪肠球菌暴发时尤其关键。还显示了在一次暴发期间,给定的MLVA图谱MT - 12(ST - 117)在五年内的稳定性。
MLVA是将粪肠球菌分离株归入不同克隆复合体的合适方法。对于调查暴发情况,当前的粪肠球菌MLVA分型方案区分能力不足,不能推荐作为优于PFGE的标准方法。