Oberhelman R A, Kopecko D J, Venkatesan M M, Salazar-Lindo E, Gotuzzo E, Yi A, Chea-Woo E, Ruiz R, Fernandez-Prada C, León-Barúa R
Department of International Health, Johns Hopkins University School of Hygiene and Public Health, Baltimore, Maryland 21205.
J Clin Microbiol. 1993 Aug;31(8):2101-4. doi: 10.1128/jcm.31.8.2101-2104.1993.
The presence of many enteropathogens which are not easily detectable by routine stool culture has led to the development of alternative diagnostic methods. One of these techniques, nucleic acid probe hybridization, has been used to identify Shigella spp. and enteroinvasive Escherichia coli (EIEC) in stool specimens through the detection of genetic material encoded by a specific large approximately 200-kbp virulence-related plasmid. In the present study, an alkaline phosphatase-labelled oligonucleotide probe developed to detect the gene for ipaH, a repetitive genetic sequence thought to be present on both the virulence-related plasmid and the chromosomes of all strains of Shigella and EIEC, was tested in a developing-country setting through a prospective clinical trial. In a group of 219 Peruvian adults and children with acute gastroenteritis, the ipaH probe detected 85% of cases of shigellosis and demonstrated a specificity of 95% when compared with simultaneous detection by several stool culture techniques. Additionally, three cases of EIEC infection which could not be diagnosed by culture methods alone were detected with the ipaH probe and were confirmed by plasmid analysis and Sereny testing. These preliminary results suggest that, with further research, the ipaH probe should prove to be a useful and rapid adjunct in the diagnosis of acute gastroenteritis in developing countries.
许多肠道病原体难以通过常规粪便培养检测出来,这促使了其他诊断方法的发展。其中一种技术,即核酸探针杂交,已被用于通过检测由一个特定的大约200千碱基对的与毒力相关的大质粒编码的遗传物质,来鉴定粪便标本中的志贺氏菌属和侵袭性大肠杆菌(EIEC)。在本研究中,一种碱性磷酸酶标记的寡核苷酸探针被开发用于检测ipaH基因,ipaH是一种重复的遗传序列,被认为存在于所有志贺氏菌和EIEC菌株的与毒力相关的质粒和染色体上。通过一项前瞻性临床试验,在一个发展中国家的环境中对该探针进行了测试。在一组219名患有急性肠胃炎的秘鲁成人和儿童中,ipaH探针检测出了85%的志贺氏菌病病例,与几种粪便培养技术同时检测相比,其特异性为95%。此外,通过ipaH探针检测出了3例仅靠培养方法无法诊断的EIEC感染病例,并通过质粒分析和塞雷尼试验得到了证实。这些初步结果表明,经过进一步研究后,ipaH探针应该会被证明是发展中国家诊断急性肠胃炎的一种有用且快速的辅助手段。