奇异变形杆菌
寻常变形杆菌
放大器
生物
聚合酶链反应
16S核糖体RNA
多重聚合酶链反应
基因
分子生物学
微生物学
DNA
遗传学
大肠杆菌
作者
А. П. Лиманский,В. В. Мінухін,О. Ю. Лиманская,N. V. Pavlenko,Марина Митрофанівна Мішина,A Tsygenenko
出处
期刊:Zhurnal mikrobiologii epidemiologii i immunobiologii
日期:2005-05-01
卷期号: (3): 33-39
被引量:4
摘要
Abstract Sets of primers for the species-specific detection of P. mirabilis and P. vulgaris by the polymerase chain reaction (PCR) were developed. As targets for these primers beta-lactamase and 16S rRNA gene fragments were chosen on the basis of the multiple leveling of the sequences of the DNA of all known P. mirabilis and P. vulgaris isolates. For differential detection oligonucleotides were selected in such a way that primers, specific for P. vulgaris, contained the non-paired nucleotide for P. mirabilis isolate at the 3'-end, and all other nucleotides were complementary to the beta-lactamase gene fragment. Primers, specific for gene 16S rRNA of P. mirabilis, contained the non-paired nucleotide for P. vulgaris isolates at the 3'-end. Standard PCR was carried out for 6 P. mirabilis and P. vulgaris strains. The use of PCR species-specific primers to P. vulgaris DNA made it possible to amplify the DNA fragment of the expected length only for P. vulgaris isolates, while the result of PCR for P. mirabilis was negative. PCR with primers specific to P. mirabilis permitted the detection of amplicon sized 101 nucleotides pairs only for P. mirabilis strains. These primers were optimized so as to use them in the specific differentiation of closely related P. mirabilis and P. vulgaris species by multiplex PCR. Genus-specific primers permitted the detection of bacterial gyrB gene of the genus Proteus were developed also.
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