Rodríguez-Angeles M G, Giono-Cerezo S, Valdespino-Gómez J L
Instituto Nacional de Diagnóstico y Referencia Epidemiológicos, México, D.F., Mexico.
Rev Latinoam Microbiol. 1994 Oct-Dec;36(4):263-71.
We studied 40 Vibrio cholerae strains: 16 from stool, 16 from sewage and 8 from food. The serotypes were Inaba in 21 strains, 8 Ogawa strains and 11 V. cholerae non-O1. PCR was made with ctx2 and ctx3 primers with 25 cycles of temperature: 1 min at 94 degrees C, 1 min at 60 degrees C and 1 min at 72 degrees C. 24 V. cholerae strains were positive: 18/24 Inaba y 6/24 Ogawa. PCR was negative for 16 strains: 3 Inaba serotype, 2 Ogawa y 11 V. cholerae non-O1. In Vero culture cells 18 strains were cytotonic, 21 cytotoxic and 1 strain was negative. ELISA was positive for 11 strains with PCR positive. The PCR sensitivity was 95.83% compared with culture cells. V. cholerae O1 produced cytotoxic effect on Vero culture cells, maybe related to ACE factor. Colony blot was made with a specific probe labeled with digoxigenin and it could detect 4 Vibrio cholerae toxigenic strains with PCR negative. All V. cholerae Non O1 strains were PCR negative.
我们研究了40株霍乱弧菌菌株：16株来自粪便，16株来自污水，8株来自食物。血清型为稻叶型的有21株，小川型8株，非O1群霍乱弧菌11株。使用ctx2和ctx3引物进行PCR，温度循环25次：94℃ 1分钟，60℃ 1分钟，72℃ 1分钟。24株霍乱弧菌菌株呈阳性：稻叶型18/24，小川型6/24。16株菌株PCR呈阴性：稻叶血清型3株，小川型2株，非O1群霍乱弧菌11株。在Vero培养细胞中，18株具有细胞膨胀性，21株具有细胞毒性，1株呈阴性。11株PCR阳性的菌株ELISA呈阳性。与培养细胞相比，PCR敏感性为95.83%。O1群霍乱弧菌对Vero培养细胞产生细胞毒性作用，可能与ACE因子有关。用洋地黄毒苷标记的特异性探针进行菌落杂交，可检测出4株PCR阴性的产毒霍乱弧菌菌株。所有非O1群霍乱弧菌菌株PCR均为阴性。
