Real-time PCR and RT-PCR comparative study of detection of dengue virus.docVIP

 PAGE \* MERGEFORMAT 14 Real-time PCR and RT-PCR comparative study of detection of dengue virus Author: Liu Jianjun, ancient Libahaer, Yang Fan, Chen Min, Ya-Qing, Yang [Abstract] Objective two kinds of methods of detection of Dengue virus, compared in order to improve detection sensitivity and specificity of dengue virus. Methods Taqman MGB technology, according to dengue virus 3 ‘non-coding region of a highly conserved sequence, dengue 1 to 4 universal fluorescent PCR primers and probes TaqmanMGB to dengue fever strain as a standard, to Japanese encephalitis strain as controls, the establishment of real-time fluorescence PCR detection of dengue virus quick. And 10 were detected by ELISA in serum samples positive RT-PCR and fluorescent PCR amplification. Results RT-PCR, 10 clinical serum samples, two were positive, the positive rate was 20%. Real-time fluorescence PCR detection of Dengue virus and Japanese encephalitis virus, no cross-reactions were detected in 10 clinical serum samples, five were positive, the positive rate was 50%. RNA extracted from the test results only 4h. Conclusion Taqman MGB real-time PCR detection method is rapid, sensitive, specific and dengue virus can be used as rapid detection method applied to the clinical early diagnosis of dengue fever. [Keywords:] dengue virus; RT-PCR; Taqman GMB Real-Time PCR [Abstract] Objective In order to improve the sensitivity and specificity of detecting dengue viruses, two methods for detecting dengue viruses were compared.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 10 serum specimens of ELISA positive were detected by the RT-PCR and fluo