[Study on the rapid detection of Dengue viruses by Taqman MGB real-time fluorescent PCR].
OBJECTIVE
To develop the Taqman MGB real-time fluorescent PCR assay for rapid detection of Dengue viruses.


METHODS
Using Taqman MGB technique, a pair of universal primers and Taqman MGB probe were designed according to a highly reserved sequence of the 3'-noncoding region of dengue viruses type 1-4. Dengue virus strains were used as standard and Japanese encephalitis virus strains were used as control, the real-time PCR assay for specific and sensitive detection of the dengue viruses was established. While 8 serum specimens of ELISA positive were detected by the RT-PCR and fluorescent PCR.


RESULTS
The sensitivity of real-time PCR was 0.17pg/microl (cDNA)or 10(-5)TCID50. There was no cross-reaction with Japanese encephalitis virus. Of 8 specimens, 2 were positive by RT-PCR and 5 were positive by real-time PCR. The test could be completed in 4 hours.


CONCLUSIONS
The Taqman MGB real-time PCR assay was fast, sensitive and specific. It could be applied to the quick early diagnosis of dengue viruses.
Liu
Jian-jun
JJ
Shenzhen Center for Disease Control and Prevention, Shenzhen 518020, China.
Yang
Fan
F
He
Jian-fan
JF
Chen
Jia-min
JM
He
Ya-qing
YQ
Yang
Hong
H
D001483:Base Sequence; D017931:DNA Primers; D003715:Dengue / Q000175:diagnosis / Q000821:virology*; D003716:Dengue Virus / Q000235:genetics / Q000302:isolation & purification*; D005456:Fluorescent Dyes; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D017931:DNA Primers; D005456:Fluorescent Dyes