Multilocus variable-number tandem repeat analysis distinguishes outbreak and sporadic Eschetichia coli O157:H7 isolates

被引:144
作者
Noller, AC
McEllistrem, MC
Pacheco, AGF
Boxrud, DJ
Harrison, LH
机构
[1] Univ Pittsburgh, Grad Sch Publ Hlth, Infect Dis Epidemiol Res Unit, Pittsburgh, PA 15261 USA
[2] Univ Pittsburgh, Sch Med, Pittsburgh, PA 15261 USA
[3] Univ Pittsburgh, Dept Infect Dis & Microbiol, Pittsburgh, PA 15261 USA
[4] Univ Pittsburgh, Dept Epidemiol, Pittsburgh, PA 15261 USA
[5] Fiocruz MS, Escola Nacl Saude Publ, Dept Epidemiol & Metodos Quantitat Suade, BR-21045900 Rio De Janeiro, Brazil
[6] Minnesota Dept Hlth, Microbiol Lab, Minneapolis, MN USA
关键词
D O I
10.1128/JCM.41.12.5389-5397.2003
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Escherichia coli O157:H7 is a major cause of food-borne illness in the United States. Outbreak detection involves traditional epidemiological methods and routine molecular subtyping by pulsed-field gel electrophoresis (PFGE). PFGE is labor-intensive, and the results are difficult to analyze and not easily transferable between laboratories. Multilocus variable-number tandem repeat (VNTR) analysis (MLVA) is a fast, portable method that analyzes multiple VNTR loci, which are areas of the bacterial genome that evolve quickly. Eighty isolates, including 21 isolates from five epidemiologically well-characterized outbreaks from Pennsylvania and Minnesota, were analyzed by PFGE and MLVA. Strains in PFGE clusters were defined as strains that differed by less than or equal to one band by using XbaI and the confirmatory enzyme SpeI. MLVA was performed by comparing the number of tandem repeats at seven loci. From 6 to 30 alleles were found at the seven loci, resulting in 64 MLVA types among the 80 isolates. MLVA correctly identified the isolates from all five outbreaks if only a single-locus variant was allowed. MLVA differentiated strains with unique PFGE types. Additionally, MLVA discriminated strains within PFGE-defined clusters that were not known to be part of an outbreak. In addition to being a simple and validated method for E. coli O157:H7 outbreak detection, MLVA appears to have a sensitivity equal to that of PFGE and a specificity superior to that of PFGE.
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页码:5389 / 5397
页数:9
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