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World J Microbiol Biotechnol. 1994 Jan;10(1):59-63. doi: 10.1007/BF00357565.
Using a 0.27 kb DNA probe specific for the heat-stable enterotoxin gene (nag-st) of Vibrio cholerae non-O1, 1109 strains representing 17 species of the genus Vibrio, isolated from clinical and environmental sources were examined. The nag-st gene was preponderantly associated with strains classified as V. mimicus; 16.8% of these strains hybridized. It was more frequent in the clinical isolates (22.6%) than in the environmental isolates (13.7%). The incidence of nag-st gene-positive strains of V. mimicus isolated from different countries was uniformly high and ranged between 8.7% (Bangladesh) and 57.1% (environmental strains from USA). The incidence of the nag-st gene was much lower among strains of V. cholerae non-O1 (3.6%). Probe-positive and-negative strains of V. mimicus and V. cholerae non-O1 were used to evaluate the performance of the conventional suckling mouse assay for detection of the NAG-ST enterotoxin. Of the 31 probe-positive strains, only five (16.1%) yielded a positive fluid accumulation ratio (FA ratio) when neat heated culture supernatant was used to perform the suckling mouse assay. All the 31 probe-positive strains gave a positive FA ratio when 20-fold concentrated and heated culture supernatants of the strains were used to perform the suckling mouse assay. The need to concentrate (by at least 20-fold) the culture supernatant of strains of V. mimicus and V. Cholerae non-O1 was identified as an important step to obtain consistent results when using the suckling mouse assay for detection of NAG-ST.
使用针对霍乱弧菌非 O1 型耐热肠毒素基因(nag-st)的 0.27kb DNA 探针,对来自临床和环境来源的 17 种弧菌属 1109 株菌进行了检测。nag-st 基因主要与被归类为 V. mimicus 的菌株相关联;其中 16.8%的菌株发生杂交。该基因在临床分离株(22.6%)中比在环境分离株(13.7%)中更为常见。从不同国家分离的 V. mimicus nag-st 基因阳性菌株的发生率均很高,范围在 8.7%(孟加拉国)至 57.1%(美国环境分离株)之间。V. cholerae non-O1 型菌株的 nag-st 基因发生率要低得多(3.6%)。使用 V. mimicus 和 V. cholerae non-O1 的探针阳性和阴性菌株来评估传统的幼鼠检测法用于检测 NAG-ST 肠毒素的性能。在 31 株探针阳性菌株中,只有 5 株(16.1%)在使用未经加热的培养上清液进行幼鼠检测时产生阳性液体蓄积比(FA 比)。当使用 20 倍浓缩和加热的菌株培养上清液进行幼鼠检测时,所有 31 株探针阳性菌株均产生阳性 FA 比。浓缩(至少 20 倍)V. mimicus 和 V. Cholerae non-O1 菌株的培养上清液被确定为在使用幼鼠检测法检测 NAG-ST 时获得一致结果的重要步骤。