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RECOMBINANT ALPHAVIRUSES ARE SAFE AND USEFUL SEROLOGICAL DIAGNOSTIC TOOLS

Serological assays for diagnosis of Venezuelan equine encephalitis virus (VEEV) currently require bio-safety level 3 facilities and select agent certification to produce antigens, reference sera, or viral stocks. Rapid identification of VEEV infection is required to respond to human and equine outbreaks of encephalitis caused by that virus and can be useful for epidemiologic surveillance. Alphavirus (Sindbis)-based recombinant viruses that express VEEV structural proteins are attenuated in animal models, thus representing an alternative to the handling of virulent infectious virus. Virus and viral antigens from recombinant Sindbis/VEE constructs engineered to express structural proteins from multiple VEEV subtypes were evaluated as diagnostic reagents in VEEV-specific serological assays, e.g., plaque reduction neutralization test (PRNT), hemagglutination inhibition (HI) assay, and complement fixation (CF) test. Chimeric viruses were produced efficiently in cell culture and were as effective as the parental virus for identifying infection of humans, horses, and rodents in these serological assays.

Received July 25, 2006. Accepted for publication September 17, 2006.

Acknowledgments: The authors thank Richard Bowen (Colorado State University, Fort Collins, CO) for providing equine serum samples and John Roehrig (Centers for Disease Control and Prevention, Fort Collins, CO) for providing monoclonal antibodies.

Financial support: Slobodan Paessler was supported by a National Institutes of Health K08 award (AI059491). This work was also supported by a grant from the National Institute of Allergy and Infectious Diseases through the Western Regional Center of Excellence for Biodefense and Emerging Infectious Diseases Research (U54 AI057156), by NIH grants awarded to S.C.W. (AI39800, AI48807) and R.B.T. (RO1-AI30027), and by an NIH contract (N01-AI25489).

^* Address correspondence to Slobodan Paessler, Department of Pathology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555-1019. E-mail: slpaessl{at}utmb.edu

Authors’ addresses: Haolin Ni, Nadezhda E. Yun, Michele A. Zacks, Scott C. Weaver, Robert B. Tesh, Amelia P. Travassos da Rosa, and Slobodan Paessler, Center for Biodefense and Emerging Infectious Diseases, Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555-1019, Telephone: +1 (409) 747-0764, Fax: +1 (409) 747-0762, E-mail: slpaessl{at}utmb.edu. Ann M. Powers, Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, CO 80522. Ilya Frolov, Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555-1019.

Reprint requests: Slobodan Paessler, Department of Pathology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555-1019, Telephone: +1 (409) 747-0764, Fax: +1 (409) 747-0762, E-mail: slpaessl{at}utmb.edu.