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English Abstract
Journal Article
[Study on the rapid detection of Dengue viruses by Taqman MGB real-time fluorescent PCR].
Wei Sheng Yan Jiu = Journal of Hygiene Research 2006 November
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.
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.
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