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Journal of Clinical Microbiology, November 1998, p. 3332-3336, Vol. 36, No. 11
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Characterization of La Crosse Virus RNA in Autopsied Central Nervous System Tissues

Laura J. Chandler,1,dagger Monica K. Borucki,1 Dawn K. Dobie,1,Dagger Leonard P. Wasieloski,1,§ Wayne H. Thompson,2 Cameron B. Gundersen,3 Kay Case,3 and Barry J. Beaty1,*

Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado 805231; Gundersen Lutheran Medical Center, La Crosse, Wisconsin 546013; and Department of Preventive Medicine, University of Wisconsin, Madison, Wisconsin 537062

Received 16 March 1998/Returned for modification 30 June 1998/Accepted 20 August 1998

A reverse transcription-PCR (RT-PCR) technique was used to detect La Crosse (LAC) virus RNA in the central nervous system (CNS) tissues of two patients who died of LAC encephalitis in 1960 and 1978. Viral RNA was readily detected by RT-PCR although the tissues had been stored frozen for up to 37 years. LAC virus was detected in the cerebral cortex but not in other CNS tissues. RT-PCR allowed detection of replicative forms of the virus, indicating that the virus was actively replicating in the specific CNS tissues. The small (S) RNA segments of the viruses from the CNS samples were demonstrated to be genetically similar by single-strand conformation polymorphism analyses. These S RNA segments were then sequenced; only two base changes were demonstrated between the 1960 and the 1978 samples, suggesting that LAC virus is genetically stable in areas of endemicity. The RT-PCR analyses of analyte directly from CNS tissues allows study of the virus without passage in cell culture.

* Corresponding author. Mailing address: Department of Microbiology, Colorado State University, Fort Collins, CO 80523. Phone: (970) 491-2988. Fax: (970) 491-1815. E-mail: Bbeaty{at}cvmbs.colostate.edu.

dagger Present address: Center for Tropical Diseases, Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555-0609.

Dagger Present address: The Neurosciences Institute, San Diego, CA 92121.

§ Present address: U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD 21702-5011.

Journal of Clinical Microbiology, November 1998, p. 3332-3336, Vol. 36, No. 11
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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