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Real-time PCR for detecting circulating dengue virus in the Guangdong Province of China in 2006

Zhijun Bai^1,, Licheng Liu^2,, Zeng Tu^2,3,, Lisi Yao⁴, Jianwei Liu¹, Bing Xu², Boheng Tang¹, Jinhua Liu¹, Yongji Wan³, Meiyu Fang¹ and Weijun Chen²

¹ Guangzhou Medical Research Institute, Yan-Ling, Guangzhou 510507, PR China

² Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 101300, PR China

³ College of Biotechnology, Southwest University, Chongqing 400715, PR China

⁴ Chinese Academy of Inspection and Quarantine, Beijing 100025, PR China

Correspondence
Meiyu Fang
fmeiyu{at}yahoo.com.cn
Weijun Chen
chenwj{at}genomics.org.cn

Received May 13, 2008
Accepted August 17, 2008

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.

Abbreviations: DF, dengue fever; DHF, dengue haemorrhagic fever; DSS, dengue shock syndrome; FAM, 6-carboxyfluorescein; IFA, immunofluorescence assay; RT-PCR, reverse transcriptase-PCR; TAMRA, 6-carboxytetramethylrhodamine; TCID₅₀, 50 % tissue culture infective dose.

These authors contributed equally to this work.

Figures showing the sensitivity and dynamic range of real-time PCR in detection of DENV RNA are available with the online version of this paper.