Application of pulsed-field gel electrophoresis for the discrimination of leptospiral isolates in Brazil
Application of pulsed-field gel electrophoresis for the discrimination of leptospiral isolates in Brazil
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DOI：
10.1111/j.1472-765x.2009.02580.x
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发表时间：
2009-05-01
影响因子：
2.4
通讯作者：
Galloway, R. L.
中科院分区：
文献类型：
--
作者：
Romero, E. C.;Blanco, R. M.;Galloway, R. L.
钩端螺旋体病是一个全球性的公共卫生问题。传统上，显微镜凝集试验（MAT）和交叉凝集素吸收试验（CAAT）是鉴别钩端螺旋体的常用方法。然而，这些技术是费力和耗时的，需要维护超过200个参考菌株和相应的兔抗血清的收集。本研究旨在探讨脉冲场凝胶电泳（PFGE）鉴别钩端螺旋体血清型的方法。对临床分离的14株钩端螺旋体进行MAT分析，然后进行PFGE鉴定。将分离株与206个钩端螺旋体参考血清型文库进行比较。所有分离株的图谱清晰，分辨率高。所有临床分离株的PFGE和MAT结果均一致。12株分离株经PFGE鉴定为血清型黄疸出血/哥本哈根型。通过MAT，这些分离物被分类为黄疸出血热血清群，滴度从3200到25600不等。两株菌株经PFGE鉴定为血清型，经MAT鉴定为血清型，滴度均大于3200。PFGE具有结果简单、可靠、重复性好等优点。PFGE为临床分离株的鉴定提供了方便的工具。
Leptospirosis is a public health problem worldwide. Traditionally, microscopic agglutination test (MAT) and cross-agglutinin absorption test (CAAT) are used to identify leptospires. However, these techniques are laborious and time-consuming, requiring the maintenance of a collection of more than 200 reference strains and correspondent rabbit antisera. The purpose of this study was to evaluate the pulsed-field gel electrophoresis (PFGE) method for discrimination of Leptospira serovars.Fourteen clinical isolates of Leptospira spp. were analysed by MAT before being characterized by PFGE. The isolates were compared with a library of 206 different reference Leptospira serovars. All the isolates gave clear profiles with high resolution. PFGE and MAT results were in agreement for all clinical isolates evaluated. Twelve isolates were classified as serovar Icterohaemorrhagiae/Copenhageni by PFGE. By MAT, these isolates were classified as serogroup Icterohaemorrhagiae with titres ranging from 3200 to 25 600. Two isolates were classified as serovar Canicola by PFGE, and as serogroup Canicola by MAT with titres higher than 3200.PFGE offers the advantages of simple, reliable and reproducible results.PFGE provides a convenient tool for the identification of clinical isolates.