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Updated: Jul 17, 2026
Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction
Published on: November 1, 2018
[Study on the rapid detection of Dengue viruses by Taqman MGB real-time fluorescent PCR]
Jian-jun Liu1, Fan Yang, Jian-fan He
1Shenzhen Center for Disease Control and Prevention, Shenzhen 518020, China.
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.
Conclusion:
The Taqman MGB real-time PCR assay was fast, sensitive and specific. It could be applied to the quick early diagnosis of dengue viruses.
