Real-time PCR for detecting circulating dengue virus in the Guangdong Province of China in 2006.
Dengue virus (DENV) causes a wide range of diseases in humans, from the acute febrile illness dengue fever (DF) to life-threatening dengue haemorrhagic fever/dengue shock syndrome. We developed four real-time quantitative PCR assays for each serotype of DENV based on computational analysis. These assays had high sensitivity and specificity without cross-reactivity for the four serotypes. To evaluate the performance of these assays in detecting and typing the virus in clinical samples, we analysed 64 serum samples from Guangdong during 2006. The results showed that 71 % of those samples were positive by the DEN-1 assay. The DENV assay results, in agreement with the serological tests and sequencing analysis, showed that the pathogen resulting in the DF explosion in Guangdong in 2006 belonged to DEN-1. Compared to the serological assays, the real-time PCR assays that we developed were much more sensitive in the 1-3 days after onset of the symptoms.
Bai
Zhijun
Z
Guangzhou Medical Research Institute, Yan-Ling, Guangzhou 510507, PR China.
Liu
Licheng
L
Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 101300, PR China.
Tu
Zeng
Z
College of Biotechnology, Southwest University, Chongqing 400715, PR China.|Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 101300, PR China.
Yao
Lisi
L
Chinese Academy of Inspection and Quarantine, Beijing 100025, PR China.
Liu
Jianwei
J
Guangzhou Medical Research Institute, Yan-Ling, Guangzhou 510507, PR China.
Xu
Bing
B
Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 101300, PR China.
Tang
Boheng
B
Guangzhou Medical Research Institute, Yan-Ling, Guangzhou 510507, PR China.
Liu
Jinhua
J
Guangzhou Medical Research Institute, Yan-Ling, Guangzhou 510507, PR China.
Wan
Yongji
Y
College of Biotechnology, Southwest University, Chongqing 400715, PR China.
Fang
Meiyu
M
Guangzhou Medical Research Institute, Yan-Ling, Guangzhou 510507, PR China.
Chen
Weijun
W
Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 101300, PR China.
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002681:China / Q000453:epidemiology; D017931:DNA Primers; D003715:Dengue / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology*; D003716:Dengue Virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012703:Serotyping; D013045:Species Specificity
D000914:Antibodies, Viral; D017931:DNA Primers; D007074:Immunoglobulin G; D007075:Immunoglobulin M