通过聚合酶链反应（PCR）对布鲁氏菌DNA进行特异性检测。
Specific detection of Brucella DNA by PCR.
作者信息
Romero C, Gamazo C, Pardo M, López-Goñi I
机构信息
Departamento de Microbiología, Universidad de Navarra, Pamplona, Spain.
出版信息
J Clin Microbiol. 1995 Mar;33(3):615-7. doi: 10.1128/jcm.33.3.615-617.1995.
Abstract
A PCR assay with primers derived from the 16S rRNA sequence of Brucella abortus was developed. Nine different combinations between six primers were tested. One pair of primers, which amplified a 905-bp fragment, was selected. As little as 80 fg of Brucella DNA was detected by this method. DNAs from all of the representative strains of the species and biovars of Brucella and from 23 different Brucella isolates were analyzed and yielded exclusively the 905-bp fragment. No amplification was detected with DNAs from 10 strains phylogenetically related to Brucella spp., 5 gram-negative bacteria showing serological cross-reactions with Brucella spp., and 36 different clinical isolates of non-Brucella species. Only Ochrobactrum anthropi biotype D yielded a PCR product of 905 bp, suggesting a closer relationship between Brucella spp. and O. anthropi biotype D. The specificity and high sensitivity of the PCR assay may provide a valuable tool for the diagnosis of brucellosis.
摘要
开发了一种基于流产布鲁氏菌16S rRNA序列引物的聚合酶链反应（PCR）检测方法。对六种引物之间的九种不同组合进行了测试。选择了一对能扩增出905bp片段的引物。该方法可检测到低至80fg的布鲁氏菌DNA。对布鲁氏菌所有代表性菌株和生物变种以及23种不同布鲁氏菌分离株的DNA进行了分析，均仅产生905bp的片段。与布鲁氏菌属在系统发育上相关的10株菌株、与布鲁氏菌属有血清学交叉反应的5株革兰氏阴性菌以及36种不同的非布鲁氏菌属临床分离株的DNA均未检测到扩增。只有嗜人苍白杆菌生物变种D产生了905bp的PCR产物，这表明布鲁氏菌属与嗜人苍白杆菌生物变种D之间关系更为密切。PCR检测方法的特异性和高灵敏度可为布鲁氏菌病的诊断提供有价值的工具。
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