Standardization and validation of the polymerase chain reaction for early diagnosis of human leptospirosis
Journal Title: Revista Peruana de Medicina Experimental y Salud Publica - Year 2007, Vol 24, Issue 1
Abstract
[i][b]Objectives: [/b][/i]To standardize and optimize a PCR assay to detect rrs gene (rRNA 16S) from Leptospira spp. to use it with samples of suspected patients of febrile syndrome, in endemic areas from Peru. [i][b]Methods:[/b][/i] We standardized and validated a PCR assay for rapid diagnostic of leptospirosis from blood and urine samples. The PCR assay was optimized with DNA samples from different species of Leptospira. Sensitivity and specificity was determined in comparison with serological test: MAT and IgM ELISA in 180 clinical samples of patients with suspect of leptopirosis. [b][i]Results:[/i][/b] The PCR assay amplified DNA of twenty five serovars of six pathogenic species of Leptospira spp. No amplification was detected with DNA from other pathogens. Sensitivity and specificity was 100% in vitro (culture samples). In blood samples, sensitivity was 100% (95CI: 97,9 - 100) in patients on the first 8 days of disease; and 30% (95CI: 16,3 - 43,7) when the time of disease was greater. In urine samples the PCR assay showed a low sensitivity. [b][i]Conclusions: [/i][/b]The standardized PCR assay for Leptospira was more sensitive than serological tests in the first days of disease. On the other hand, it has low sensitivity when bacterial population is poor.
Authors and Affiliations
Manuel Céspedes, Rafael Tapia, Lourdes Balda, Dana González, Carlos Peralta, Patricia Condori
Keywords
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