Ooteman Márcia Costa, Vago Annamaria Ravara, Koury Matilde Cota
Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
Can J Microbiol. 2004 Dec;50(12):1073-9. doi: 10.1139/w04-096.
In this study we tested the potential use of low-stringency single specific primer-PCR (LSSP-PCR) for genetically typing Leptospira directly from biological samples. Serum samples obtained from 29 patients with clinically suspected leptospirosis were amplified by specific PCR, using the previously selected G1 and G2 primers. The PCR products of approximately 300 bp were subsequently used as a template for LSSP-PCR analysis. We were able to produce genetic signatures from the leptospires present in the human samples, which permitted us to make a preliminary identification of the infective serovar by comparing the LSSP-PCR profiles obtained directly from serum samples with those from reference leptospires. Thus, LSSP-PCR has the potential to become a useful diagnostic tool for identifying leptospires in biological samples without the need for bacteria isolation and culture.
在本研究中，我们测试了低严谨度单特异性引物聚合酶链反应（LSSP-PCR）直接从生物样本中对钩端螺旋体进行基因分型的潜在用途。使用先前选定的G1和G2引物，通过特异性聚合酶链反应对从29例临床疑似钩端螺旋体病患者获得的血清样本进行扩增。随后，将约300 bp的聚合酶链反应产物用作LSSP-PCR分析的模板。我们能够从人类样本中存在的钩端螺旋体产生基因特征，通过将直接从血清样本获得的LSSP-PCR图谱与参考钩端螺旋体的图谱进行比较，使我们能够对感染血清型进行初步鉴定。因此，LSSP-PCR有潜力成为一种有用的诊断工具，用于在无需细菌分离和培养的情况下鉴定生物样本中的钩端螺旋体。
