用于产肠毒素大肠杆菌的碱性磷酸酶和放射性标记基因探针与生物测定法的比较。

A comparison of alkaline phosphatase and radiolabelled gene probes with bioassays for enterotoxigenic Escherichia coli.

作者信息

Bopp C A, Threatt V L, Moseley S L, Wells J G, Wachsmuth I K

机构信息

Division of Bacterial Diseases, Centers for Disease Control, Atlanta, Georgia 30333.

出版信息

Mol Cell Probes. 1990 Jun;4(3):193-203. doi: 10.1016/0890-8508(90)90053-3.

DOI:10.1016/0890-8508(90)90053-3

PMID:2199821

Abstract

Alkaline phosphatase-conjugated oligonucleotide probes (APO), 32P-labelled oligonucleotide (RO) and cloned polynucleotide (RP) probes were evaluated for their ability to detect enterotoxigenic Escherichia coli (ETEC) as defined by bioassay. These three sets of probes were applied to 301 E. coli strains that had previously been defined by the Y1 adrenal cell assay for heat-labile enterotoxin (LT) and the infant mouse assay for heat-stable enterotoxin (ST). The correlation of the APO probe for LT with the bioassay was 98% with five discrepancies and a positive predictive value (PPV) of 100%. For the APO/ST probe the correlation with the bioassay was 98% with seven discrepancies and a PPV of 96%. The correlation of the RO probe for LT was 99% with four discrepancies and a PPV of 100%, while the overall correlation for the two RO/ST probes was 97% with eight discrepancies and a PPV of 97%. For the RP probes, the correlation for LT was 99% with four discrepancies and a PPV of 100% and for ST was 98% with seven discrepancies and a PPV of 98%. These findings suggest that the APO probes were as sensitive as the RO and RP probes in detecting ETEC by colony hybridization and could be a practical alternative to bioassays and radiolabelled probes for ETEC since they do not require expensive equipment or extensive technical training.

摘要

评估了碱性磷酸酶偶联的寡核苷酸探针（APO）、32P 标记的寡核苷酸（RO）和克隆的多核苷酸（RP）探针通过生物测定法检测产肠毒素大肠杆菌（ETEC）的能力。这三组探针应用于 301 株大肠杆菌，这些菌株先前已通过 Y1 肾上腺细胞测定法检测热不稳定肠毒素（LT）以及通过幼鼠测定法检测热稳定肠毒素（ST）来进行定义。APO 探针检测 LT 与生物测定法的相关性为 98%，有 5 个差异，阳性预测值（PPV）为 100%。对于 APO/ST 探针，与生物测定法的相关性为 98%，有 7 个差异，PPV 为 96%。RO 探针检测 LT 的相关性为 99%，有 4 个差异，PPV 为 100%，而两种 RO/ST 探针的总体相关性为 97%，有 8 个差异，PPV 为 97%。对于 RP 探针，检测 LT 的相关性为 99%，有 4 个差异，PPV 为 100%，检测 ST 的相关性为 98%，有 7 个差异，PPV 为 98%。这些发现表明，APO 探针在通过菌落杂交检测 ETEC 方面与 RO 和 RP 探针一样敏感，并且对于 ETEC 而言，它们可以成为生物测定法和放射性标记探针的实用替代方法，因为它们不需要昂贵的设备或广泛的技术培训。