Li Yayue, Liu Dan, Cao Boyang, Han Weiqing, Liu Yanqun, Liu Fenxia, Guo Xi, Bastin David A, Feng Lu, Wang Lei
TEDA School of Biological Sciences and Biotechnology, Nankai University, 23 Hong Da Street, TEDA, Tianjin 300457, China.
J Clin Microbiol. 2006 Dec;44(12):4376-83. doi: 10.1128/JCM.01389-06. Epub 2006 Oct 4.
Shigella and pathogenic Escherichia coli are major causes of human infectious diseases and are responsible for millions of cases of diarrhea worldwide every year. A convenient and rapid method to identify highly pathogenic serotypes of Shigella and E. coli is needed for large-scale epidemiologic study, timely clinical diagnosis, and reliable quarantine of the pathogens. In this study, a DNA microarray targeting O-serotype-specific genes was developed to detect 15 serotypes of Shigella and E. coli, including Shigella sonnei; Shigella flexneri type 2a; Shigella boydii types 7, 9, 13, 16, and 18; Shigella dysenteriae types 4, 8, and 10; and E. coli O55, O111, O114, O128, and O157. The microarray was tested against 186 representative strains of all Shigella and E. coli O serotypes, 38 clinical isolates, and 9 strains of other bacterial species that are commonly present in stool samples and was shown to be specific and reproducible. The detection sensitivity was 50 ng genomic DNA or 10(4) CFU per ml in mock stool specimens. This is the first report of a microarray for serotyping Shigella and pathogenic E. coli. The method has a number of advantages over traditional bacterial culture and antiserum agglutination methods and is promising for applications in basic microbiological research, clinical diagnosis, food safety, and epidemiological surveillance.
志贺氏菌和致病性大肠杆菌是人类传染病的主要病因,每年在全球导致数百万例腹泻病例。大规模流行病学研究、及时的临床诊断以及对病原体进行可靠的检疫都需要一种便捷快速的方法来鉴定志贺氏菌和大肠杆菌的高致病性血清型。在本研究中,开发了一种针对O-血清型特异性基因的DNA微阵列,用于检测15种志贺氏菌和大肠杆菌血清型,包括宋内志贺氏菌、2a型福氏志贺氏菌、7型、9型、13型、16型和18型鲍氏志贺氏菌、4型、8型和10型痢疾志贺氏菌以及大肠杆菌O55、O111、O114、O128和O157。该微阵列针对所有志贺氏菌和大肠杆菌O血清型的186株代表性菌株、38株临床分离株以及粪便样本中常见的9株其他细菌物种进行了测试,结果显示具有特异性和可重复性。在模拟粪便标本中的检测灵敏度为50 ng基因组DNA或每毫升10(4) CFU。这是关于志贺氏菌和致病性大肠杆菌血清分型微阵列的首次报道。该方法相对于传统细菌培养和抗血清凝集方法具有许多优势,在基础微生物学研究、临床诊断、食品安全和流行病学监测中具有应用前景。