This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Chappell, W. A. Articles by Henderson, B. E. Search for Related Content PubMed Articles by Chappell, W. A. Articles by Henderson, B. E. Agricola Articles by Chappell, W. A. Articles by Henderson, B. E.

 Previous Article  |  Next Article 

Appl Environ Microbiol. 1971 December; 22(6): 1100-1103
Copyright © 1971 American Society for Microbiology. All Rights Reserved.

Comparison of Three Methods Used to Isolate Dengue Virus Type 2

Center for Disease Control, Atlanta, Georgia 30333

ABSTRACT

During the 1969 dengue epidemic in Puerto Rico, human sera and Aedes aegypti mosquitoes were collected for virus isolation and identification. Three methods of isolation were used and compared. In the first method, we inoculated newborn mice by the intracranial route, noted any signs of illness, and serially passed specimens in mice until virus was isolated. In the second method, we inoculated tube cultures of LLC-MK₂ cells, noted any cytopathic effect (CPE), and assayed fluids for virus by plaque formation in LLC-MK₂ cell monolayers. The third method was different from the second only in that the original specimens were first inoculated into fluid cultures of Singh's A. albopictus cells. No significant CPE was seen in LLC-MK₂ cultures; however, distinct syncytial CPE was observed in A. albopictus cells. About the same number of virus isolates were made in each isolation system. Virus isolates from both sera and mosquitoes were identified as dengue type 2 by a plaque-reduction neutralization test in LLC-MK₂ cells. The utility of the three methods, individually or in combination, is discussed and related to diagnostic and epidemic situations.

¹ Present address: 2281 66th Avenue, S.E., Mercer Island, Wash. 98040.

² Present address: Department of Pathology, School of Medicine, University of California, Los Angeles, Calif. 90033.