Ross Ian L, Parkinson Ian H, Heuzenroeder Michael W
Infectious Diseases Laboratories, Institute of Medical and Veterinary Science, Adelaide, SA, Australia.
Int J Med Microbiol. 2009 Jan;299(1):37-41. doi: 10.1016/j.ijmm.2008.05.009. Epub 2008 Jul 9.
The resolving power of multiple-locus variable-number tandem repeat analysis (MLVA) was undertaken on 78 phenotypically closely related isolates of Salmonella enterica serovar Typhimurium that were previously analysed with multiple amplification of phage locus typing (MAPLT) and pulsed-field gel electrophoresis (PFGE). Isolates were tested with 10 primer sets targeting tandem repeat loci in the S. Typhimurium genome. A new primer set targeting the SB21(ST64B) locus was also assessed for MAPLT analysis. Both methods produced similar levels of discrimination between 41 non-DT 126 isolates while MLVA proved superior in separating the DT 126 isolates. Results showed that two loci, STTR5 and STTR6, provided the most variation in tandem repeat numbers between the 78 S. Typhimurium isolates with 11 alleles detected for each locus. Some isolates did not produce amplified product with PCR for various loci, providing another level for discrimination. The remaining MLVA loci provided less allelic variation, with 4 loci failing to provide any discrimination. There were a number of isolates that were shown to be unique by one method but were clustered by the other method. Combining the most variable regions of both MAPLT and MLVA into one assay may provide significant resolution of isolates with the minimal number of primers utilised.
对78株表型密切相关的肠炎沙门氏菌鼠伤寒血清型菌株进行了多位点可变数目串联重复序列分析(MLVA)的分辨能力研究,这些菌株之前已通过噬菌体位点分型多重扩增(MAPLT)和脉冲场凝胶电泳(PFGE)进行了分析。用10组引物对鼠伤寒沙门氏菌基因组中的串联重复位点进行检测。还评估了一组针对SB21(ST64B)位点的新引物用于MAPLT分析。两种方法在41株非DT126菌株间产生了相似的分辨水平,而MLVA在区分DT126菌株方面表现更优。结果显示,STTR5和STTR6这两个位点在78株鼠伤寒沙门氏菌菌株间的串联重复数变化最大,每个位点检测到11个等位基因。一些菌株对不同位点的PCR未产生扩增产物,这提供了另一种分辨水平。其余的MLVA位点等位基因变异较少,有4个位点没有提供任何分辨能力。有许多菌株通过一种方法显示为独特的,但通过另一种方法则聚类在一起。将MAPLT和MLVA最具变异性的区域组合到一个检测中,可能用最少数量的引物对菌株提供显著的分辨能力。