Rasschaert G, Houf K, Imberechts H, Grijspeerdt K, De Zutter L, Heyndrickx M
Ghent University, Faculty of Veterinary Medicine, Department of Veterinary Public Health and Food Safety, Salisburylaan 133, 9820 Merelbeke, Belgium.
J Clin Microbiol. 2005 Aug;43(8):3615-23. doi: 10.1128/JCM.43.8.3615-3623.2005.
Five repetitive-element PCR (rep-PCR) techniques [primer sets ERIC1R-ERIC2 and REP1R-REP2I and primers ERIC2, BOXA1R, and (GTG)5] were evaluated for the discrimination of Salmonella enterica isolates at the serotype level. On the basis of number, even distribution over the whole fingerprint, and clarity of bands in the fingerprints, the enterobacterial repetitive intergenic consensus (ERIC) primer set and the (GTG)5 primer were chosen for use in the following experiments. For these two primer sets, reproducibility was tested on different lysates of five selected serotypes of Salmonella in the same PCR by using three different PCR runs. Reproducibility was poor between different PCR runs but high within the same PCR run. Furthermore, 80 different serotypes and five isolates which were not typeable by serotyping were fingerprinted. All strains were typeable by the ERIC primer set and the (GTG)5 primer and generated unique fingerprints, except for some strains with incomplete antigenic codes. Finally, 55 genetically different strains belonging to 10 serotypes were fingerprinted to examine the genetic diversity of the rep-PCR within serotypes. This experiment showed that one serotype did not always correlate to only one ERIC or (GTG)5 fingerprint but that the fingerprint heterogeneity within a serotype was limited. In epidemiological studies, ERIC- and/or (GTG)5-PCR can be used to limit the number of strains that have to be serotyped. The reproducibility of isolates in one PCR run, the discriminatory power, and the genetic diversity (stability) of the fingerprint were similar for the Eric primer set and the (GTG)5 primer, so both are equally able to discriminate Salmonella serotypes.
评估了五种重复元件PCR（rep-PCR）技术[引物对ERIC1R-ERIC2和REP1R-REP2I以及引物ERIC2、BOXA1R和(GTG)5]用于在血清型水平区分肠炎沙门氏菌分离株的能力。基于条带数量、在整个指纹图谱上的均匀分布以及指纹图谱中条带的清晰度，选择肠杆菌重复基因间共有序列（ERIC）引物对和(GTG)5引物用于后续实验。对于这两种引物对，通过三次不同的PCR扩增，在同一PCR中对五种选定血清型的沙门氏菌的不同裂解物进行了重复性测试。不同PCR扩增之间的重复性较差，但在同一PCR扩增内重复性较高。此外，对80种不同血清型以及5株无法通过血清分型鉴定的分离株进行了指纹图谱分析。除了一些抗原编码不完整的菌株外，所有菌株均可通过ERIC引物对和(GTG)5引物进行分型，并产生独特的指纹图谱。最后，对属于10种血清型的55株遗传不同的菌株进行了指纹图谱分析，以研究血清型内rep-PCR的遗传多样性。该实验表明，一种血清型并不总是只与一种ERIC或(GTG)5指纹图谱相关，而是血清型内的指纹图谱异质性有限。在流行病学研究中，ERIC-和/或(GTG)5-PCR可用于减少需要进行血清分型的菌株数量。ERIC引物对和(GTG)5引物在一次PCR扩增中分离株的重复性、鉴别能力以及指纹图谱的遗传多样性（稳定性）相似，因此两者在区分沙门氏菌血清型方面同样有效。
