Evaluation of PCR-Based Identification and Genotyping Methods for the iap Genes in Listeria monocytogenes and other Listeria spp.
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概要
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By using 181 <I>Listeria</I> spp. strains except for <I>L. ivanovii</I>, we evaluated the three distinct PCR methods targeting the <I>iap</I> genes encoding p60 proteins for genus or species identification, and the PCR-RFLP analysis for genotyping. Each of the two species-specific PCR methods yielded the expected amplicons from all of the 126 <I>L. monocytogenes</I> and 43 <I>L. innocua</I> strains tested. The genus-specific PCR method amplified the target regions (about 1.4 kb) from 178 <I>Listeria</I> strains other than3 <I>L. grayi</I> strains tested. The PCR-RFLP analysis with either <I>HhaI</I> or <I>Hind</I>III showed distinct several patterns: 97<I>L. monocytogenes</I> strains divided into 2 genotypes (types I and II) with <I>Hin</I>d III and 5 genotypes (types A, B, C, D and E) with<I>Hha</I>I. On the other hand, 43 <I>L. innocua</I> strains showed an identical genotype (type I with <I>Hin</I>d III and type F with<I>Hha</I>l). Bot h <I>L. seeligeri</I> and <I>L. welshimeri</I> strains showed a distinct genotype (type I with<I>Hin</I>d III and type G with<I>Hha</I>l). The <I>L. monocytogenes</I> genotypes correlated with the serovars. All 4b strains exhibited one genotype (type I and type A). 1/2b strains exhibited type I and types A or B. In contrast, 1/2a strains exhibited type II and types C, D, or E. 1/2c strains exhibited only type II and type C. Thus, these results suggest that the PCR and PCR-RFLP methods can identify or estimate the genus and species of listerial strains rapidly and are useful tools to genotype <I>Listeria</I> spp. strains phylogenetically.
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