Detection and typing of dengue virus using polymerase chain reaction and microwell plate hybridization.
- Author:
Rui-wen REN
1
;
Xiao-li XU
;
Jian-jun LI
;
Mei-yu FANG
;
Jian-wei LIU
;
An-de MA
Author Information
- Publication Type:Journal Article
- MeSH: Dengue Virus; classification; genetics; Enzyme-Linked Immunosorbent Assay; methods; Nucleic Acid Hybridization; methods; Polymerase Chain Reaction; methods; Reproducibility of Results; Serotyping; methods
- From: Journal of Southern Medical University 2006;26(9):1356-1362
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a specific, sensitive and practicable method for detection and typing of dengue virus.
METHODSBased on the genomic sequence analysis of dengue virus types 1-4, 4 pairs of primers were designed. The specific capture probes of dengue virus types 1-4 were amplified using RT-PCR, cloned and sequenced before using them for precoating the microwell plate. The samples were amplified using biotin-labeled forward primer and reverse primer, and microwell plate hybridization was carried out for detection and typing of dengue virus types 1-4.
RESULTSThe absorbance of the positive samples were higher than 0.5, while the average absorbance of the negative samples was lower than 0.1, with the S/N higher than 10.
CONCLUSIONThe method of PCR-ELISA we established for early detection and typing of all 4 dengue viruses seretypes.
