PCR-RESTRICTION FRAGMENT LENGTH POLYMORPHISM ANALYSIS OF THE OSPC GENE FOR DETECTION OF MIXED CULTURE AND FOR EPIDEMIOLOGIC TYPING OF BORRELIA-BURGDORFERI SENSU-STRICTO

Restriction fragment length polymorphism (RFLP) analysis of the outer surface protein C (ospC) gene amplicon was used for rapid screening fo r genetic variability within Borrelia burgdorferi sensu stricto specie s and for detection of multiple borreliae in culture. Primers for the ospC gene amplified a fragment of about 600 bp from Borrelia cultures, After cleavage of the amplified products by MboI and DraI, eight diff erent RFLP types were found among 13 B. burgdorferi sensu stricto stra ins from various sources and geographical areas, and three RFLP types were found among 10 representative isolates from skin biopsy specimens taken from patients residing on the eastern end of Long Island, New Y ork (B. W. Berger, R. C. Johnson, C. Kodner, and L. Coleman, J. Clin. Microbiol. 30:359-361, 1992). These results suggested that the DNA org anization of B. burgdorferi sensu stricto is heterogeneous not only gl obally but also within a localized geographical area and that the ospC -based typing approach could differentiate the B. burgdorferi sensu st ricto. From the results obtained using mixed cultures of two different RFLP types of B. burgdorferi sensu stricto, contamination of at least 0.5% of different types of Borrelia cells in culture could be detecte d. This method could detect a multiple-B. burgdorferi sensu stricto in fection in the bladders of mice experimentally infected with two diffe rent RFLP type strains. The present study showed that RFLP analysis of ospC-PCR products is a reliable method for epidemiological typing of B. burgdorferi sensu stricto and could be used for rapid detection of mixed Borrelia culture and multiple B. burgdorferi sensu stricto infec tions in animals, ticks, and patients.