Oprandy J J, Thornton S A, Gardiner C H, Burr D, Batchelor R, Bourgeois A L
Infectious Diseases Department, Naval Medical Research Institute, Bethesda, Maryland 20814.
J Clin Microbiol. 1988 Jan;26(1):92-5. doi: 10.1128/jcm.26.1.92-95.1988.
Several studies have demonstrated the usefulness of 32P-labeled recombinant DNA probes for identifying enterotoxigenic Escherichia coli (ETEC). The use of radioisotopes and X-ray development, however, severely handicaps the utility of DNA probes in most clinical laboratories. In this study, enzyme-labeled oligonucleotide probes for ETEC LT (heat-labile toxin) and ST (heat-stable toxin) genes were compared with the standard Y1 adrenal cell and suckling mouse assays for their ability to identify ETEC in a population of American adults experiencing acute episodes of diarrhea in South America and West Africa. The LT probe hybridized with 12% (64 of 529) of the E. coli colonies tested, whereas 11% (57 of 529) were positive by Y1 adrenal cell assay. DNA from 9% (47 of 529) of the E. coli colonies tested hybridized with the ST probe, whereas only 5% (28 of 529) produced ST as measured by the suckling mouse bioassay. For the patient samples tested, correlation between probe and bioassay for LT was 97%, or three discrepancies in 111 patients tested. Overall concordance of the ST probe and bioassay was 95%, or five discrepancies in 111 patients. Enzyme-labeled oligonucleotide probes represent a major advance in the diagnosis of ETEC-associated diarrheal disease and may be used in laboratories with minimal equipment.
多项研究已证明32P标记的重组DNA探针在鉴定产肠毒素大肠杆菌(ETEC)方面的实用性。然而,放射性同位素和X射线显影的使用严重限制了DNA探针在大多数临床实验室中的应用。在本研究中,将针对ETEC LT(热不稳定毒素)和ST(热稳定毒素)基因的酶标记寡核苷酸探针与标准的Y1肾上腺细胞和乳鼠试验进行比较,以评估它们在南美洲和西非经历急性腹泻发作的美国成年人人群中鉴定ETEC的能力。LT探针与12%(529个测试的大肠杆菌菌落中的64个)的大肠杆菌菌落杂交,而通过Y1肾上腺细胞试验检测为阳性的比例为11%(529个中的57个)。9%(529个测试的大肠杆菌菌落中的47个)的大肠杆菌菌落DNA与ST探针杂交,而通过乳鼠生物测定法检测仅5%(529个中的28个)产生ST。对于测试的患者样本,LT探针与生物测定法之间的相关性为97%,即在111名测试患者中有3个差异。ST探针与生物测定法的总体一致性为95%,即在111名患者中有5个差异。酶标记寡核苷酸探针代表了ETEC相关性腹泻疾病诊断的一项重大进展,并且可在设备最少的实验室中使用。