Sethabutr O, Hanchalay S, Echeverria P, Taylor D N, Leksomboon U
Lancet. 1985 Nov 16;2(8464):1095-7. doi: 10.1016/s0140-6736(85)90687-7.
A non-radioactive biotinylated DNA probe was constructed to detect Shigella and enteroinvasive Escherichia coli (EIEC). Specimens were examined with the biotinylated probe after removing streptavidin-binding glycoproteins with proteinase K. Both biotinylated and radioactive probes detected 125 pg of target-cell DNA after hybridisation for 24 h and exposure to indicator dyes or X-ray film for 4 h. Both probes hybridised with 52 EIEC and none of 16 non-EIEC examined; they also hybridised with stool blots from 11 of 13 children with culture-proven shigellosis or EIEC diarrhoea and were negative with stool blots from 43 children who were culture negative for Shigella and EIEC. Biotinylated DNA probes can be as sensitive as radiolabelled probes, but have the advantage of a longer shelf-life and greater availability.
构建了一种非放射性生物素化DNA探针,用于检测志贺氏菌和侵袭性大肠杆菌(EIEC)。在用蛋白酶K去除链霉亲和素结合糖蛋白后,用生物素化探针检查标本。生物素化探针和放射性探针在杂交24小时并暴露于指示染料或X射线胶片4小时后,均能检测到125 pg的靶细胞DNA。两种探针均与52株EIEC杂交,而与所检测的16株非EIEC均不杂交;它们还与13名经培养证实患有志贺氏菌病或EIEC腹泻的儿童中的11名儿童的粪便印迹杂交,而与43名志贺氏菌和EIEC培养阴性的儿童的粪便印迹呈阴性。生物素化DNA探针的灵敏度可与放射性标记探针相同,但具有保质期更长和更易获得的优点。