Echeverria P, Seriwatana J, Taylor D N, Tirapat C, Chaicumpa W, Rowe B
J Infect Dis. 1985 Jan;151(1):124-30. doi: 10.1093/infdis/151.1.124.
Radioactively labeled enterotoxin genes were used to study the epidemiology of enterotoxigenic Escherichia coli infections in two Thai villages. When E. coli that were isolated from 674 specimens were fixed on nitrocellulose paper and examined for hybridization with E. coli enterotoxin gene probes in Bangkok, the technique had a sensitivity of 94% (31 of 33) and a specificity of 100% (641 of 641), when compared with tests of E. coli for enterotoxin production in the Y-1-adrenal cell and suckling-mouse assays. However, when the same specimens were fixed directly onto nitrocellulose paper at a field laboratory and transported to the reference laboratory for assay with the gene probes, 27 specimens that contained enterotoxigenic E. coli did not hybridize with the E. coli gene probes. Enterotoxigenic E. coli that hybridized with the LT, ST-H, and ST-P probes were identified in 10% (17 of 177) of villagers with diarrhea, 7% (8 of 108) of contacts of individuals with diarrhea caused by enterotoxigenic E. coli, and 3% (32 of 1,199) of persons not associated with cases of diarrhea caused by enterotoxigenic E. coli. Enterotoxigenic E. coli that hybridized with the ST-II probe was not a cause of diarrhea. Alternative methods of retaining DNA on filters under field conditions are needed before this technique can be used for direct examination of specimens with enterotoxin gene probes.
利用放射性标记的肠毒素基因研究了泰国两个村庄产肠毒素大肠杆菌感染的流行病学情况。当将从674份标本中分离出的大肠杆菌固定在硝酸纤维素纸上,并在曼谷检测其与大肠杆菌肠毒素基因探针的杂交情况时,与在Y-1肾上腺细胞和乳鼠试验中检测大肠杆菌产生肠毒素的试验相比,该技术的灵敏度为94%(33份中的31份),特异性为100%(641份中的641份)。然而,当在现场实验室将相同标本直接固定在硝酸纤维素纸上,然后运至参考实验室用基因探针进行检测时,27份含有产肠毒素大肠杆菌的标本未与大肠杆菌基因探针杂交。与LT、ST-H和ST-P探针杂交的产肠毒素大肠杆菌在10%(177名腹泻村民中的17名)腹泻村民、7%(108名产肠毒素大肠杆菌所致腹泻患者接触者中的8名)产肠毒素大肠杆菌所致腹泻患者接触者以及3%(1199名与产肠毒素大肠杆菌所致腹泻病例无关的人员中的32名)与产肠毒素大肠杆菌所致腹泻病例无关的人员中被鉴定出来。与ST-II探针杂交的产肠毒素大肠杆菌不是腹泻的病因。在该技术可用于用肠毒素基因探针直接检测标本之前,需要找到在现场条件下将DNA保留在滤膜上的替代方法。