Robins-Browne R M, Miliotis M D, Cianciosi S, Miller V L, Falkow S, Morris J G
Department of Microbiology, University of Melbourne, Parkville, Victoria, Australia.
J Clin Microbiol. 1989 Apr;27(4):644-50. doi: 10.1128/jcm.27.4.644-650.1989.
The virulence of yersiniae varies according to (i) species and biotype and (ii) possession of a 67- to 72-kilobase virulence plasmid. Y. pestis, Y. pseudotuberculosis, and biotypes 1B, 2, 3, 4, and 5 of Y. enterocolitica are inherently virulent but express full virulence only when in possession of a virulence plasmid. Other Yersinia species and biotypes 1A and 3B of Y. enterocolitica are seldom implicated in disease. In this study, we prepared DNA probes from eight nonoverlapping regions of the virulence plasmid of a strain of Y. enterocolitica and from the inv and ail chromosomal loci responsible for the invasive capacity of Y. enterocolitica and Y. pseudotuberculosis. The probes were used in colony hybridization experiments to investigate 156 yersiniae of various species and biotypes and of differing virulence. Probes prepared from the inv gene of Y. pseudotuberculosis hybridized with Y. pseudotuberculosis and Y. pestis only, whereas an analogous probe prepared from Y. enterocolitica hybridized with all species and biotypes of yersiniae (but not with other bacteria) regardless of virulence or potential virulence. Probes prepared from the ail region of Y. enterocolitica reacted almost exclusively with Y. enterocolitica strains of pathogenic biotypes. Probes prepared from the virulence plasmid of a serogroup O:8, biotype 1B isolate of Y. enterocolitica identified virulent yersiniae in all species with a high degree of sensitivity and specificity. These probes did not react with yersiniae of avirulent biotypes or species. Of the other assays of virulence evaluated (calcium dependence, binding of crystal violet, and pyrazinamidase activity), binding of crystal violet provided a simple means for identifying plasmid-bearing strains.
（i）菌种和生物型，以及（ii）是否拥有67至72千碱基的毒力质粒。鼠疫耶尔森氏菌、假结核耶尔森氏菌以及小肠结肠炎耶尔森氏菌的生物型1B、2、3、4和5具有内在毒力，但只有在拥有毒力质粒时才会表达出完全毒力。其他耶尔森氏菌种以及小肠结肠炎耶尔森氏菌的生物型1A和3B很少与疾病相关。在本研究中，我们从一株小肠结肠炎耶尔森氏菌毒力质粒的八个非重叠区域以及负责小肠结肠炎耶尔森氏菌和假结核耶尔森氏菌侵袭能力的inv和ail染色体位点制备了DNA探针。这些探针用于菌落杂交实验，以研究156株不同菌种、生物型和毒力的耶尔森氏菌。从假结核耶尔森氏菌的inv基因制备的探针仅与假结核耶尔森氏菌和鼠疫耶尔森氏菌杂交，而从小肠结肠炎耶尔森氏菌制备的类似探针与所有菌种和生物型的耶尔森氏菌（但不与其他细菌）杂交，无论其毒力或潜在毒力如何。从小肠结肠炎耶尔森氏菌的ail区域制备的探针几乎只与致病生物型的小肠结肠炎耶尔森氏菌菌株反应。从O:8血清群、生物型1B的小肠结肠炎耶尔森氏菌分离株的毒力质粒制备的探针以高度的敏感性和特异性鉴定了所有菌种中的有毒力耶尔森氏菌。这些探针不与无毒力生物型或菌种的耶尔森氏菌反应。在评估的其他毒力测定方法（钙依赖性、结晶紫结合和吡嗪酰胺酶活性）中，结晶紫结合为鉴定携带质粒的菌株提供了一种简单方法。
