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用于快速常规鉴定沙门氏菌的沙门氏菌特异性生物素化DNA探针的研制。

Development of a Salmonella-specific biotinylated DNA probe for rapid routine identification of Salmonella.

作者信息

Gopo J M, Melis R, Filipska E, Meneveri R, Filipski J

机构信息

University of Zimbabwe, Biological Sciences Department, Harare.

出版信息

Mol Cell Probes. 1988 Dec;2(4):271-9. doi: 10.1016/0890-8508(88)90011-4.

Abstract

Classical microbiological techniques used in the detection and identification of Salmonella spp. in foods, drinking water and clinical samples are relatively lengthy. Immunoassays, on the other hand, have the major disadvantage of often generating false positives and false negatives. Recombinant DNA technology offers more efficient alternatives to the detection of a specific organism by employing cloned DNA sequences unique to the organism. Demonstration of a presence of complementary sequences among a heterogeneous population of molecules of DNA isolated from bacteria can be made by using a DNA-DNA hybridization technique. We have obtained a fragment of DNA from Salmonella typhimurium chromosomal DNA, cloned it in Escherichia coli plasmid and tested it in colony hybridization tests with 57 strains of Salmonella and other enterobacteriaceae. In all tests, the fragment was found to be Salmonella-specific in that it gave a positive reaction with all strains of Salmonella tested and was negative when tested against other Enterobacteriaceae.

摘要

用于检测和鉴定食品、饮用水及临床样本中沙门氏菌属的传统微生物技术相对耗时。另一方面,免疫测定的主要缺点是经常产生假阳性和假阴性结果。重组DNA技术通过使用特定生物体独有的克隆DNA序列,为检测特定生物体提供了更高效的替代方法。通过DNA-DNA杂交技术,可以证明从细菌中分离出的异质DNA分子群体中存在互补序列。我们从鼠伤寒沙门氏菌染色体DNA中获得了一段DNA片段,将其克隆到大肠杆菌质粒中,并在菌落杂交试验中对57株沙门氏菌和其他肠杆菌科细菌进行了测试。在所有测试中,该片段被发现具有沙门氏菌特异性,因为它与所有测试的沙门氏菌菌株都产生了阳性反应,而与其他肠杆菌科细菌测试时呈阴性。

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