Chetina E V, Gintsburg A L, Grizhebovskiĭ G M, Briukhanov A F, Zigangirova N A, Popov V D, Kurbanov Sh Kh
Zh Mikrobiol Epidemiol Immunobiol. 1992 Mar(3):21-5.
A specific method of the isolation of the cholera toxin gene by the directional amplification of DNA in the polymerase chain reaction (PCR) has been developed. The product of this reaction has a molecular weight of 440 sequence pairs and is a DNA fragment located on the A-subunit of V. cholerae gene vct. The sensitivity of the method permits the detection of one bacterial cell in the reaction mixture. The method is effective when V. cholerae purified DNA, cell lysates and the DNA of total microflora isolated from the water of natural springs are used. The study of water samples from natural water bodies by the method of PRC has revealed cholera toxin genes of V. cholerae noncultivated forms ni 5 out of 7 water samples taken from natural water bodies at the regions of Azerbaijan endemic for cholera and made it possible to evaluate the number of V. cholerae. The prospects of using PCR for the control of the epidemiological situation in regions endemic for cholera are discussed.
已开发出一种通过聚合酶链反应(PCR)中DNA的定向扩增来分离霍乱毒素基因的特定方法。该反应产物的分子量为440个序列对,是位于霍乱弧菌基因vct A亚基上的DNA片段。该方法的灵敏度允许检测反应混合物中的一个细菌细胞。当使用霍乱弧菌纯化DNA、细胞裂解物以及从天然泉水分离的总微生物群落的DNA时,该方法有效。通过PCR方法对天然水体水样的研究表明,在阿塞拜疆霍乱流行地区采集的7份天然水体水样中,有5份检测到了未培养形式的霍乱弧菌霍乱毒素基因,并使得评估霍乱弧菌数量成为可能。文中还讨论了使用PCR控制霍乱流行地区流行病学状况的前景。