Preston M A, Borczyk A A
Clinical Bacteriology Section, Ministry of Health, Toronto, Ontario, Canada.
J Clin Microbiol. 1994 Jun;32(6):1427-30. doi: 10.1128/jcm.32.6.1427-1430.1994.
Restriction fragment length polymorphisms (RFLPs) of genomic DNAs from 49 clinical isolates of Shigella sonnei were analyzed by using a modified restriction endonuclease analysis procedure to investigate the genetic variability of this species. After cleavage with the restriction enzyme HaeIII or RsaI, DNA samples were electrophoresed in polyacrylamide gels and the RFLP patterns were visualized by silver staining. The results showed that among 20 strains associated with sporadic cases of infection in three Canadian provinces, 15 distinct RFLP patterns were revealed by HaeIII digestion and 12 distinct patterns were revealed by RsaI digestion. In contrast, the RFLP patterns of individual isolates within six groups of epidemiologically related isolates were identical to each other but distinct from those of unrelated isolates, and these patterns could be used to determine the genetic relationships between isolates associated with separate outbreaks of shigellosis. Our results indicate that the modified restriction endonuclease analysis technique represents a rapid, reproducible, and highly discriminatory method for the molecular typing of this species.
采用改良的限制性内切酶分析方法，对49株宋内志贺菌临床分离株的基因组DNA进行限制性片段长度多态性（RFLP）分析，以研究该菌的遗传变异性。用限制性内切酶HaeIII或RsaI切割后，将DNA样品在聚丙烯酰胺凝胶中进行电泳，通过银染观察RFLP图谱。结果显示，在来自加拿大三个省份的20株散发病例相关菌株中，HaeIII酶切显示出15种不同的RFLP图谱，RsaI酶切显示出12种不同的图谱。相比之下，6组流行病学相关分离株中各分离株的RFLP图谱彼此相同，但与无关分离株的图谱不同，这些图谱可用于确定与志贺菌病不同暴发相关的分离株之间的遗传关系。我们的结果表明，改良的限制性内切酶分析技术是一种快速、可重复且具有高度鉴别力的该菌分子分型方法。
