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Utility and Limitations of Direct Multi-Locus Sequence Typing on qPCR-Positive Blood to Determine Infecting Leptospira Strain

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  • Department of Community Medicine, Faculty of Medicine and Allied Sciences, Rajarata University of Sri Lanka, Saliyapura, Sri Lanka; Division of Infectious Diseases, Department of Medicine, University of California at San Diego, La Jolla, California

Culture-independent molecular characterization of infecting Leptospira human blood specimens from a 2008 outbreak of human leptospirosis in central Sri Lanka was carried out. Of 58 quantitative real-time polymerase chain reaction-positive samples analyzed for seven multi-locus sequence typing (MLST) housekeeping genes (mreA, pfkB, pntA, sucA, tpiA, fadD, and glmU), interpretable data was obtained from 12 samples. Mean bacterial load was 2.2 × 105 among specimens with complete MLST profiles compared with 1.3 × 104 among specimens without complete MLST profiles; all specimens with complete profiles had at least 4.9 × 104 Leptospira/mL (t = 5, P < 0.001). Most (11/12) identified sequence types were ST1 (L. interrogans serovar Lai) and ST44 (L. interrogans serovar Geyaweera). MLST can be used to directly identify infecting Leptospira strains in blood samples obtained during acute illness without the need for culture isolation, but it shows important limitations related to bacterial load.

Author Notes

* Address correspondence to Joseph M. Vinetz, University of California at San Diego, School of Medicine, Center for Tropical Medicine and Emerging Infectious Diseases, Division of Infectious Diseases, Department of Medicine, 9500 Gilman Drive 0741, Palade Laboratories, Room 125, La Jolla, CA 92093-0741. E-mail: jvinetz@ucsd.edu

Authors' addresses: Suneth B. Agampodi, Department of Community Medicine, Faculty of Medicine and Allied Sciences, Rajarata University of Sri Lanka, Saliyapura, Sri Lanka, E-mail: sunethagampodi@yahoo.com. Angelo C. Moreno, Graduate Program in Molecular Genetics and Microbiology, Duke University, Durham, NC, E-mail: angelo.moreno@duke.edu. Joseph M. Vinetz, Department of Medicine, School of Medicine, University of California at San Diego, La Jolla, CA, E-mail: jvinetz@ucsd.edu. Michael A. Matthias, Division of Infectious Diseases, Department of Medicine, University of California at San Diego, School of Medicine, La Jolla, CA, E-mail: mmatthias@ucsd.edu.

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