Evaluation of nested polymerase chain reaction for the early detection of Leptospira spp. DNA in serum samples from patients with leptospirosis

• Published in: Diagnostic microbiology and infectious disease Vol. 78; no. 4; pp. 343 - 346
• Main Authors: Blanco, Roberta Morozetti, Romero, Eliete Caló
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-04-2014
• Subjects: DNA, Bacterial - genetics; DNA, Bacterial - isolation & purification; Early Diagnosis; Humans; Infectious Disease; Internal Medicine; Leptospira - genetics; Leptospira - isolation & purification; Leptospira spp; Leptospirosis; Leptospirosis - diagnosis; Molecular diagnosis; Molecular Diagnostic Techniques - methods; Nested PCR; Polymerase Chain Reaction - methods; Prospective Studies; Serum; Serum - microbiology; Leptospirosis; Serum; Nested PCR; Molecular diagnosis; Leptospira spp
• ISSN: 0732-8893; 1879-0070
• DOI: 10.1016/j.diagmicrobio.2013.12.009

Abstract The aim of this study was to analyse the nested polymerase chain reaction (nested PCR) in human serum samples of patients with clinical manifestations of leptospirosis. The cases of leptospirosis were defined by the microagglutination test (MAT). The samples were collected in 2010. Of 1042 serum samples collected from 521 patients, 28 (5.4%) were considered positive cases of leptospirosis, and 493 (94.6%) were negative. Twenty-three confirmed cases had no MAT-detectable antibodies in the acute sample (mean of 5.6 days after onset). Nested PCR was positive in 22/23 (95.7%) patients during the acute phase of the disease, with negative results by MAT. Nested PCR was negative in all convalescent serum samples with positive results by MAT. All negative cases of leptospirosis were negative by nested PCR. The nested PCR is an alternative diagnostic tool for early detection of leptospires in sera during the first 7 days of the disease.