Potential application of low-stringency single specific primer-PCR in the identification of Leptospira in the serum of patients with suspected leptospirosis.
In this study we tested the potential use of low-stringency single specific primer-PCR (LSSP-PCR) for genetically typing Leptospira directly from biological samples. Serum samples obtained from 29 patients with clinically suspected leptospirosis were amplified by specific PCR, using the previously selected G1 and G2 primers. The PCR products of approximately 300 bp were subsequently used as a template for LSSP-PCR analysis. We were able to produce genetic signatures from the leptospires present in the human samples, which permitted us to make a preliminary identification of the infective serovar by comparing the LSSP-PCR profiles obtained directly from serum samples with those from reference leptospires. Thus, LSSP-PCR has the potential to become a useful diagnostic tool for identifying leptospires in biological samples without the need for bacteria isolation and culture.
Ooteman
Márcia Costa
MC
Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
Vago
Annamaria Ravara
AR
Koury
Matilde Cota
MC
D015373:Bacterial Typing Techniques; D001431:Bacteriological Techniques; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000032:analysis; D006801:Humans; D007919:Leptospira / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D007922:Leptospirosis / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D044967:Serum / Q000382:microbiology*
D004269:DNA, Bacterial