Publication: Applications of Polymerase Chain Reaction for Identification of Dengue Viruses Isolated from Patient Sera
Issued Date
1993-01-01
Resource Type
ISSN
13480421
03855600
03855600
Other identifier(s)
2-s2.0-0027511654
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Mahidol University
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SCOPUS
Bibliographic Citation
Microbiology and Immunology. Vol.37, No.1 (1993), 774-777
Suggested Citation
Virat Sirisanthana, Kouichi Morita, Mariko Tanaka, Akira Igarashi, K. kul, Bruce L. Innis, Ananda Nisalak, N. Nimmanitya, Niwat Maneekarn, Wipawee Usawattana-, Nopporn Sittisombut, S. Suchitra, Akira Igarashi Applications of Polymerase Chain Reaction for Identification of Dengue Viruses Isolated from Patient Sera. Microbiology and Immunology. Vol.37, No.1 (1993), 774-777. doi:10.1111/j.1348-0421.1993.tb03177.x Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/22598
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Title
Applications of Polymerase Chain Reaction for Identification of Dengue Viruses Isolated from Patient Sera
Abstract
A simple and sensitive procedure of reverse transcriptase polymerase chain reaction (RT-PCR) was developed previously such that all 4 serotypes of dengue viruses could be detected and their serotypes identified simultaneously in a single-step procedure. In this study we compared the RT-PCR with a conventional immunoperoxidase (PAP) staining method for the identification of dengue viruses currently isolated from patient sera. Sixty-six sera taken from dengue hemorrhagic fever (DHF) patients were subjected to virus isolation by inoculating onto C6/36 cell cultures. Screening for the presence of dengue viruses in culture fluids was done after 7 days of incubation by PAP staining using hyperimmune rabbit anti-dengue virus antibody as the primary reagent. Dengue viruses in positive cultures were further identified for their serotypes by PAP using type-specific monoclonal antibodies (MAb) and by RT-PCR. Thirty-two out of the 66 serum specimens tested (48. 5%) were positive for dengue viruses. of these, 5 were type 1 (DEN-1), 25 were type 2 (DEN-2) and 2 contained both DEN-1 and DEN-2. All cultures that were positive by PAP method were also positive by RT-PCR and vice versa. Thus, the results obtained by RT-PCR were in good agreement with those by PAP. It is important to point out that while all 5 DEN-1 isolates reacted readily with the MAb 1F1, only 2 of them could be identified by the MAb 15F3. Our data suggest that antigenic variation among DEN-1 isolates occur frequently and this should be taken into consideration in the selection of appropriate type-specific MAb for serotyping of dengue viruses. We also demonstrated dual infection of DEN-1 and DEN-2 in two patients’ sera by RT-PCR. © 1993, Center For Academic Publications Japan. All rights reserved.