Echeverria P, Taylor D N, Seriwatana J, Brown J E, Lexomboon U
Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand.
J Clin Microbiol. 1989 Feb;27(2):331-4. doi: 10.1128/jcm.27.2.331-334.1989.
We compared three methods for detecting enteropathogens in 416 children with diarrhea: (i) examination of 10 lactose-fermenting and all non-lactose-fermenting Escherichia coli (colony blots); (ii) examination of 300 colonies (replicate blots); and (iii) determination of the total bacterial growth of stools (stool blots). All specimens were spotted onto Whatman 541 filters and hybridized with specific radiolabeled DNA probes. Enterotoxigenic E. coli was detected in 38 patients by examining colony blots, in 52 patients by examining replicate blots, and in 45 patients by examining stool blots. Enteropathogenic E. coli adhesin factor was detected in 12 patients by colony blots, in 25 patients by replicate blots, and in 16 patients by stool blots. E. coli that hybridized with the enterohemorrhagic E. coli probe was detected in 2 patients by colony blots, in 11 patients by replicate blots, and in 0 patients by stool blots. Shiga-like toxin-producing E. coli was detected in 0 patients by colony blots, in 12 patients by replicate blots, and in 0 patients by stool blots. Shigella spp. were identified by standard bacteriological methods in 82 patients, and enteroinvasive E. coli was identified by colony blots in 11 patients (total, 93), by replicate blots in 56 patients, and by stool blots in 35 patients. Of 82 culture-confirmed Shigella infections, 45 were identified by examining replicate blots with the 17-kilobase-pair probe and 36 were identified by examination with the Ipa probe (P less than 0.05). Examining replicate blots with specific probes identified more enterotoxigenic E.coli (P < 0.005), enteropathogenic E.coli adhesion factor-producing E.coli (P < 0.001), and Shiga-like toxin-producing E.coli (P < 0.005) infections than examining colony blots. More Shigella and enteroinvasive E.coli infections were identified by standard bacteriological methods and examining colony blots with a specific probe than by examining replicate and stool blots (P < 0.001).
我们比较了三种检测416例腹泻儿童肠道病原体的方法：（i）检查10株乳糖发酵型和所有非乳糖发酵型大肠杆菌（菌落印迹法）；（ii）检查300个菌落（重复印迹法）；（iii）测定粪便中的细菌总生长量（粪便印迹法）。所有标本均点样于Whatman 541滤膜上，并用特异性放射性标记DNA探针进行杂交。通过菌落印迹法在38例患者中检测到产肠毒素大肠杆菌，通过重复印迹法在52例患者中检测到，通过粪便印迹法在45例患者中检测到。通过菌落印迹法在12例患者中检测到致病性大肠杆菌黏附因子，通过重复印迹法在25例患者中检测到，通过粪便印迹法在16例患者中检测到。与肠出血性大肠杆菌探针杂交的大肠杆菌，通过菌落印迹法在2例患者中检测到，通过重复印迹法在11例患者中检测到，通过粪便印迹法在0例患者中检测到。通过菌落印迹法在0例患者中检测到产志贺样毒素大肠杆菌，通过重复印迹法在12例患者中检测到，通过粪便印迹法在0例患者中检测到。通过标准细菌学方法在82例患者中鉴定出志贺菌属，通过菌落印迹法在11例患者中鉴定出侵袭性大肠杆菌（共93例），通过重复印迹法在56例患者中鉴定出，通过粪便印迹法在35例患者中鉴定出。在82例经培养确诊的志贺菌感染中，45例通过用17千碱基对探针检查重复印迹法鉴定出，36例通过用Ipa探针检查鉴定出（P<0.05）。与检查菌落印迹法相比，用特异性探针检查重复印迹法鉴定出更多的产肠毒素大肠杆菌（P<0.005）、产致病性大肠杆菌黏附因子大肠杆菌（P<0.001）和产志贺样毒素大肠杆菌（P<0.005）感染。与检查重复印迹法和粪便印迹法相比，通过标准细菌学方法和用特异性探针检查菌落印迹法鉴定出更多的志贺菌和侵袭性大肠杆菌感染（P<0.001）。
