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Journal of Clinical Microbiology, September 2001, p. 3303-3310, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3303-3310.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Evaluation of Recombinant Leptospira Antigen-Based Enzyme-Linked Immunosorbent Assays for the Serodiagnosis of Leptospirosis

Brendan Flannery,1 Dirceu Costa,2 Fernanda Pinheiro Carvalho,2 Hygia Guerreiro,2,3 James Matsunaga,4,5 Emilson Domingos Da Silva,6 Antonio Gomes Pinto Ferreira,6 Lee W. Riley,1 Mitermayer G. Reis,2 David A. Haake,4,5 and Albert I. Ko2,7,*

School of Public Health, University of California, Berkeley, California 947201; Gonçalo Moniz Research Center, Oswaldo Cruz Foundation, Brazilian Ministry of Health,2 and School of Pharmacy, Federal University of Bahia,3 Salvador, Brazil; Division of Infectious Diseases, Veterans Affairs Greater Los Angeles Healthcare System, Los Angeles, California 900734; Department of Medicine, UCLA School of Medicine, Los Angeles, California 900955; Biomanguinhos, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Rio de Janeiro, Brazil6; and Division of International Medicine and Infectious Disease, Weill Medical College of Cornell University, New York, New York 100217

Received 26 February 2001/Returned for modification 28 May 2001/Accepted 1 July 2001

There is an urgent need for development of new serodiagnostic strategies for leptospirosis, an emerging zoonosis with worldwide distribution. We have evaluated the diagnostic utility of five recombinant antigens in enzyme-linked immunosorbent assays (ELISAs) for serodiagnosis of leptospirosis. Sera from 50 healthy residents of a high-incidence region were used to determine cutoff values for 96% specificity. In paired sera from 50 cases of leptospirosis confirmed by the microscopic agglutination test, immunoglobulin G (IgG) but not IgM reacted with the recombinant leptospiral proteins. The recombinant LipL32 IgG ELISA had the highest sensitivities in the acute (56%) and convalescent (94%) phases of leptospirosis. ELISAs based on recombinant OmpL1, LipL41, and Hsp58 had sensitivities of 16, 24, and 18% during the acute phase and 72, 44, and 32% during convalescence, respectively. Compared to sera from healthy individuals, patient sera did not react significantly with recombinant LipL36 (P > 0.05). Recombinant LipL32 IgG ELISA demonstrated 95% specificity among 100 healthy individuals, and specificities ranging from 90 to 97% among 30 dengue patients, 30 hepatitis patients, and 16 patients with diseases initially thought to be leptospirosis. Among 39 Venereal Disease Research Laboratory test-positive individuals and 30 Lyme disease patients, 13 and 23% of sera, respectively, reacted positively with the rLipL32 antigen. These findings indicate that rLipL32 may be an useful antigen for the serodiagnosis of leptospirosis.

* Corresponding author. Mailing address: Centro de Pesquisas Gonçalo Moniz, Fundação Oswaldo Cruz, Rua Waldemar Falcão, 121, 40295-001 Salvador, Brazil. Phone: (55 71) 356-4320, ext. 243. Fax: (55 71) 356-2155. E-mail: aik2001{at}med.cornell.edu.

Journal of Clinical Microbiology, September 2001, p. 3303-3310, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3303-3310.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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