[Detection and typing of dengue virus using polymerase chain reaction and microwell plate hybridization].
OBJECTIVE
To establish a specific, sensitive and practicable method for detection and typing of dengue virus.


METHODS
Based 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.


RESULTS
The 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.


CONCLUSIONS
The method of PCR-ELISA we established for early detection and typing of all 4 dengue viruses seretypes.
Ren
Rui-wen
RW
Disease Control and Prevention Center of Guangzhou Command, Guangzhou 510507, China. renruiwen@hotmail.com
Xu
Xiao-li
XL
Li
Jian-jun
JJ
Fang
Mei-yu
MY
Liu
Jian-wei
JW
Ma
An-de
AD
D003716:Dengue Virus / Q000145:classification / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012703:Serotyping / Q000379:methods