大理地区志贺菌的血清型分布及多重PCR检测技术的应用

目的 探讨大理地区志贺菌的血清型分布以及多重PCR技术在志贺菌检测中的应用.方法 用常规分离培养、生化鉴定和血清学分型法对腹泻患者粪标本进行检测,分离到的标本再用多重PCR法对志贺菌的毒力基因shetA、shetB、ial和ipaH进行扩增检测.结果 通过常规培养和生化反应鉴定,得到68株志贺菌.血清学鉴定显示福氏志贺菌41株,宋内志贺菌25株,鲍氏志贺菌2株.通过多重PCR扩增后68株志贺菌均在423 bp(ipaH)处出现了条带,检出率为100%,其余在147 bp(shetB)、309 bp(shetA)处以及320 bp(ial)处均出现了至少一个条带,与常规鉴定法的符合率达100%.5份直接用阳性粪便标本提取DNA为模板进行多重PCR扩增后,也得到了同样的结果.结论 大理地区细菌性痢疾患者感染以福氏志贺菌为主,多重PCR技术可用于志贺菌的快速检测和流行病学调查.

Study on serotype distribution in Dali and application of multiplex PCR assay for shigella detection

Objective To study serotype distribution in Dali and application of multiplex PCR assay for shigel- la detection. Methods 68 shigella were tested by traditional culture, biochemical identification and serological typing first. Then all samples were detected by multiplex PCR assay including our virulence genes which were shetA,shetB, ial and ipaft. Results There were 68 isolated from acute diarrhea faeces after identified by traditional culture. Sero logical assay classified as follows：41strains were Shigella flexneri,25 strains were Shigella sonnei,2 stains were Shi- gella bogdii. All the 68 stains shigellosis appeared line in 423 bp（ipaH） ,and at least appeared another line in 147 bp （shetB）or 309 bp（sketA）or 320 bp（ial）by multiplex PCR assay. The coincidence rate was 100% comparing tradition al culture. It was the same result with the method of the DNA plate extracted directly from feces by multiplex PCR assay. Conclusion The main serotypes of shigella isolates in Dali are Shigella flexneri. The multiplex PCR assay can rapidly detect shigella.