Molecular characterization of Leptospira sp. strains isolated from human subjects in São Paulo, Brazil using a polymerase chain reaction-based assay: a public health tool
Molecular characterization of Leptospira sp. strains isolated from human subjects in São Paulo, Brazil using a polymerase chain reaction-based assay: a public health tool
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DOI：
10.1590/s0074-02762006000400005
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发表时间：
2006-06-01
期刊：
影响因子：
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通讯作者：
Yasuda, Paulo H
中科院分区：
文献类型：
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作者：
Romero, Eliete C;Yasuda, Paulo H
聚合酶链反应（PCR）为基础的检测，扩增细菌基因组内存在的重复DNA元件被用来表征和区分钩端螺旋体属。35株参考培养物收集和18个临床分离株，其中以前已分析交叉凝集素吸收试验（CAAT）进行了评价，通过这种技术。参考培养物采集分析的PCR结果显示，没有对应于血清群Australis、Autumnalis、Bataviae、Celledoni、Cynopteri、Djasiman、Panama、Pomona、Pyrogenes和Tarassovi的条带。然而，PCR方法能够清楚地区分Andamana、Ballum、犬型、流感伤寒型、七日型、黄疸出血型、爪哇型、Sejroe、Semaranga和Shermani血清群。先前CAAT鉴定为Copenhageni血清型、Castellonis血清型和犬型血清型的临床分离株与PCR结果一致。以前的临床分离株的特点是血清型波莫纳没有分化的PCR。还通过该PCR方法评价了从巴西圣保罗获得的来自钩端螺旋体病患者的另外40个临床分离株。其中39例被确定属于黄疸出血群（97.5%），1例属于Seiroe群（2.5%）。结果表明，本研究所建立的PCR方法可用于钩端螺旋体的血清型水平的快速分型，并可用于流行病学调查。
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 Sao 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 Seiroe (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.