Romero Eliete C, Yasuda Paulo H
Laboratório de Leptospirose, Divisão de Biologia Médica, Instituto Adolfo Lutz, São Paulo, SP, 01246-902, Brasil.
Mem Inst Oswaldo Cruz. 2006 Jun;101(4):373-8. doi: 10.1590/s0074-02762006000400005.
A polymerase chain reaction (PCR)-based assay which amplifies repetitive DNA elements present within bacterial genomes was used to characterize and differentiate Leptospira sp. Thirty-five strains from a reference culture collection and 18 clinical isolates which had been previously analyzed by cross agglutinin absorption test (CAAT) were evaluated by this technique. PCR results from analysis of the reference culture collection showed no bands corresponding to serogroups Australis, Autumnalis, Bataviae, Celledoni, Cynopteri, Djasiman, Panama, Pomona, Pyrogenes, and Tarassovi. However, the PCR method was able to clearly discriminate the serogroups Andamana, Ballum, Canicola, Grippotyphosa, Hebdomadis, Icterohaemorrhagiae, Javanica, Sejroe, Semaranga, and Shermani. Clinical isolates previously characterized by CAAT as serovar Copenhageni, serovar Castellonis, and as serovar Canicola were in agreement with PCR results. The clinical isolate previously characterized as serovar Pomona was not differentiated by PCR. Forty additional clinical isolates from patients with leptospirosis obtained in São Paulo, Brazil were also evaluated by this PCR method. Thirty-nine of these were determined to belong to serogroup Icterohaemorrhagiae (97.5%) and one to serogroup Sejroe (2.5%). These results demonstrate that the PCR method described in this study has utility for rapid typing of Leptospira sp. at the serogroup level and can be used in epidemiological survey.
一种基于聚合酶链反应（PCR）的检测方法被用于对钩端螺旋体属进行特征描述和鉴别，该方法可扩增细菌基因组中存在的重复DNA元件。对来自参考培养物保藏中心的35株菌株以及18株先前已通过交叉凝集素吸收试验（CAAT）分析过的临床分离株，采用该技术进行了评估。对参考培养物保藏中心的分析结果显示，PCR产物中没有与澳洲群、秋季群、巴达维亚群、细胞多尼群、犬蝠群、贾西曼群、巴拿马群、波摩那群、致热群和塔拉索夫群相对应的条带。然而，PCR方法能够清晰地区分安达马纳群、拜伦群、犬型群、七日热群、七日热群、出血性黄疸群、爪哇群、 sejroe群、Semaranga群和Shermani群。先前通过CAAT鉴定为哥本哈根血清型、卡斯泰洛尼血清型和犬型血清型的临床分离株，其结果与PCR结果一致。先前鉴定为波摩那血清型的临床分离株，PCR未能将其区分。另外还采用该PCR方法对从巴西圣保罗的钩端螺旋体病患者中获得的40株临床分离株进行了评估。其中39株被确定属于出血性黄疸群（97.5%），1株属于sejroe群（2.5%）。这些结果表明，本研究中描述的PCR方法在钩端螺旋体属血清群水平的快速分型方面具有实用性，可用于流行病学调查。
