Low-stringency single specific primer PCR for identification of Leptospira
Low-stringency single specific primer PCR for identification of Leptospira
复制标题
DOI：
10.1099/jmm.0.04923-0
复制
发表时间：
2003-02-01
影响因子：
3
通讯作者：
Koury, MC
中科院分区：
文献类型：
--
作者：
do Oliveira, MAA;Caballero, OL;Koury, MC
通过低严格单特异性引物 PCR (LSSP-PCR) 技术对问号钩端螺旋体、borgpetersenii 钩端螺旋体、santarosai 钩端螺旋体、kirschneri 钩端螺旋体、韦氏钩端螺旋体、双折钩端螺旋体和 meyeri 钩端螺旋体的 35 种钩端螺旋体血清型进行了表征。进行 LSSP-PCR 分析以检测使用 G1 和 G2 选择的引物从基因组 DNA 扩增的 285 bp DNA 片段中的 DNA 多态性。来自相同基因组物种的血清型获得了类似的 LSSP-PCR 谱，而来自非相关物种的血清型则产生了不同的多带模式。根据序列分析数据，用钩端螺旋体不同血清型的G1和G2引物扩增的所有基因组片段长度均为285 bp，不同基因组物种之间观察到的核苷酸变异最常见。 LSSP-PCR 技术的简单性和准确性被发现适合于钩端螺旋体物种的鉴定。
Thirty-five Leptospira serovars from the species Leptospira interrogans, Leptospira borgpetersenii, Leptospira santarosai, Leptospira kirschneri, Leptospira weilii, Leptospira biflexa and Leptospira meyeri were characterized by the low-stringency single specific primer PCR (LSSP-PCR) technique. LSSP-PCR analysis was performed to detect DNA polymorphisms in a 285 bp DNA fragment amplified from genomic DNA with G1 and G2 selected primers. Similar LSSP-PCR profiles were obtained for serovars from the same genomic species, while serovars from non-related species produced distinct multiband patterns. Based on the data from sequence analysis, all genomic fragments amplified with G1 and G2 primers from distinct serovars of Leptospira were 285 bp in length, with nucleotide variation observed most frequently among different genomic species. The simplicity and accuracy of the LSSP-PCR technique were found to be suitable for identification of Leptospira species.