| 郭喜玲,史智扬,顾玲,庄菱,潘浩.应用多重引物PCR技术检测O157∶H7毒力基因[J].Chinese journal of Epidemiology,2000,21(6):410-412 |
| 应用多重引物PCR技术检测O157∶H7毒力基因 |
| Using multiplex PCR for the detection of virulence genes in Escherichia coli O157∶H7 |
| Received:June 28, 2000 |
| DOI： |
| KeyWord: 肠出血性大肠杆菌 基因 聚合酶链反应 |
| English Key Word: Enterohemorrhagic E.coli(EHEC) Gene Polymerase chain reaction |
| FundProject: |
| Author Name | Affiliation | | GUO Xiling | Antiepidemic Station of Jiangsu Province, Nanjing 210009, China | | SHI Zhiyang | Antiepidemic Station of Jiangsu Province, Nanjing 210009, China | | GU Ling | Antiepidemic Station of Jiangsu Province, Nanjing 210009, China | | ZHANG Ling | Antiepidemic Station of Jiangsu Province, Nanjing 210009, China | | PAN Hao | Antiepidemic Station of Jiangsu Province, Nanjing 210009, China |
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| Abstract: |
| 目的 对江苏省 6个不同地区不同宿主动物中分离的O15 7∶H7菌株进行毒力基因的检测分析。方法 应用肠出血性大肠杆菌 (EHEC)的多重引物聚合酶链反应 (PCR)方法,以志贺样毒素 (SLT2 和SLT1 )基因、“粘附抹平”因子eaeA基因和溶血素 (hly)基因为靶基因进行检测。结果 江苏省分离的O15 7∶H7菌株毒力基因携带率为 5 6.5 %,不同地区的分离株携带率有所不同,个别地区高达 90 %以上,有的地区则未检测到带毒力基因的菌株,这一结果与不同地区发病率的高低有平行的关系。疾病高发地区,菌株毒力基因携带率达 85.7% (36 /4 2 ),低发或散发地区为 5 2.6 % (10 /19),非流行地区为 8.3 % (2 /2 4)。不同的宿主动物分离株其毒力基因携带阳性率从高到低依次为羊 >牛 >猪>鸡。仅有的一份兔粪便标本分离株也检出毒力基因。菌株毒力基因图谱以SLT2 +eaeA +hly为主,占 79.2 %,其次为SLT2 +SLT1+eaeA +hly和SLT2 +hly,分别占 16.6 %和 4.2 % ；有毒力基因的菌株均有hly和SLT2,绝大多数菌株有eaeA基因,携带SLT1 的菌株则较少,这与国外一些报道有所不同。结论 O15 7∶H7毒力基因图谱是一个重要的分子流行病学标志,应用多重引物PCR方法检测O15 7∶H7毒力基因,简便、快速、特异、敏感,对流行病学调查分析 |
| English Abstract: |
| Objective To detect and characterize the virulence genes in E.coli O157∶H7 isolated from various reserv oir in six areas of Jiang su province.Method The virulence genes of Shiga-like tox in(SLT1 and SLT2 ), intimin(eaeA)and hemoly sin(hlyA)were chosen as the target genes and amplified in multiplex PCR assay s.Results Of the eighty-five E.coli O157∶H7 strains, the overall virulence g ene prevalence w as found to be 56.5 %(48/ 85).The prevalence rates viralence genes of isolates from various areas were different from 0 % up to 90.5 %.It seemed to ex ist a relationship betw een the virulence g eneprevalence and the level of incidence.In the areas w here rates of incidence were divided into high, low,sporadic or zero, the prev alence rates were 85.7 %(36/ 42), 52.6 %(10/19) and 8.3 %(2/ 24),respectively.The prev alence rates of isolates were also different from various reservoirs, decrcasing by sheep, cattle, pig and poultry.One isolate from a rabbit w as positive for SLT2, eaeA and hly genes.Of for ty-eight isolates car ry ing virulence genes, 38(79.2 %) had SLT2, eaeA and hly genes, taking the dominate virulence g ene pattern, 8(16.6 %)had all of the four virulence genes 2(4.2%)had both SLT2 and hly genes respectively.In addition, SLT1 gene showed a low er prev alence, which was different from some findings abroad.Conclusion Since virulence gene patter n of E.coli O157∶H7 is an important molecular epidemiolo gical marker, it can provide an useful information for epidemiologic studies, and helpful to the desig n of prevention and co ntrol strategies.For virulence gene de tection, multiplex PCR seems to be a simple, rapid, specific and sensitive method. |
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