Han Weiqing, Liu Bin, Cao Boyang, Beutin Lothar, Krüger Ulrike, Liu Hongbo, Li Yayue, Liu Yanqun, Feng Lu, Wang Lei
TEDA School of Biological Sciences and Biotechnology, Nankai University, 23 Hongda Street, TEDA, Tianjin 300457, China.
Appl Environ Microbiol. 2007 Jun;73(12):4082-8. doi: 10.1128/AEM.01820-06. Epub 2007 Apr 20.
Escherichia coli strains causing postweaning diarrhea (PWD) and edema disease (ED) in pigs are limited to a number of serogroups, with O8, O45, O138, O139, O141, O147, O149, and O157 being the most commonly reported worldwide. In this study, a DNA microarray based on the O-antigen-specific genes of all 8 E. coli serogroups, as well as 11 genes encoding adhesion factors and exotoxins associated with PWD and ED, was developed for the identification of related serogroups and virulence gene patterns. The microarray method was tested against 186 E. coli and Shigella O-serogroup reference strains, 13 E. coli reference strains for virulence markers, 43 E. coli clinical isolates, and 12 strains of other bacterial species and shown to be highly specific with reproducible results. The detection sensitivity was 0.1 ng of genomic DNA or 10(3) CFU per 0.3 g of porcine feces in mock samples. Seventeen porcine feces samples from local hoggeries were examined using the microarray, and the result for one sample was verified by the conventional serotyping methods. This microarray can be readily used to screen for the presence of PWD- and ED-associated E. coli in porcine feces samples.
引起仔猪断奶后腹泻(PWD)和水肿病(ED)的大肠杆菌菌株仅限于多个血清群,其中O8、O45、O138、O139、O141、O147、O149和O157是全球最常报道的血清群。在本研究中,基于所有8种大肠杆菌血清群的O抗原特异性基因,以及11种编码与PWD和ED相关的粘附因子和外毒素的基因,开发了一种DNA微阵列,用于鉴定相关血清群和毒力基因模式。该微阵列方法针对186株大肠杆菌和志贺氏菌O血清群参考菌株、13株大肠杆菌毒力标记参考菌株、43株大肠杆菌临床分离株以及12株其他细菌物种进行了测试,结果显示具有高度特异性且结果可重复。在模拟样品中,检测灵敏度为每0.3克猪粪便0.1纳克基因组DNA或10³CFU。使用该微阵列对当地养猪场的17份猪粪便样品进行了检测,其中一个样品的结果通过传统血清分型方法进行了验证。这种微阵列可方便地用于筛查猪粪便样品中与PWD和ED相关的大肠杆菌的存在。