Leptospirosis diagnosis using Nested-PCR

Brazilian Journal of Microbiology

Fernanda Nassi¹  Fabiana Kömmling Seixas¹  Sandra Denize Dorneles Jouglard¹  Simone Simionatto¹  Everton Fagonde Silva¹  Núbia Seyffert¹  Claudiomar Soares Brod¹  Odir Antonio Dellagostin¹ 

[1] ,Universidade Federal de Pelotas Centro de Biotecnologia Pelotas RS ,Brasil

关键词: Leptospirosis;    PCR;    Leptospirose;    PCR;   

DOI  :  10.1590/S1517-83822003000500031

来源: SciELO

【 摘 要 】

Leptospirosis is a worldwide sanitary problem. Its clinical signs resemble that of other diseases like Dengue and Flu, and it is difficult to distinguish between them. Currently available diagnostic methods shown low sensitivity and specificity. Efforts have been made to develop simpler, faster and more efficient diagnostic methods. The aim of this work was to evaluate and optimize a Nested-PCR method for diagnosis of leptospirosis. Primers were designed to amplify a 264 bp region within the LipL32 gene. The sensitivity and specificity of the assay was evaluated using seven saprophytic serovars and 35 pathogenic serovars. This technique showed to be very specific for pathogenic serovars, however it lacked sensitivity. In order to enhance the sensitivity, another primer pair was designed which amplify a 183 bp region within the 264 bp region in lipL32 gene, and used in a Nested-PCR assay. This approach was much more sensitive than traditional PCR.

【 授权许可】

CC BY-NC   
 All the contents of this journal, except where otherwise noted, is licensed under a Creative Commons Attribution License