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Antigen sandwich elisa predicts RT-PCR detection of dengue virus genome in infected culture fluids of aedes albopictus C6/36 cells.

Author

Corazon C. Buerano,
Filipinas F. Natividad,
Rodolfo C. Contreras,
Ima Nursa Ibrahim,
Marlou Noel M. Mangada,
Futoshi Hasebe,
Shingo Inoue,
Ronald R. Matias,
Akira Igarashi

Related Institution

Institute of Biology, College of Science, University of the Philippines

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Publication Information

Publication Type
Journal
Publication Sub Type
Journal Article, Original
Title
St. Luke's Journal of Medicine
Frequency
Semi-Annual
Publication Date
January-June 2010
Volume
6
Issue
1
Page(s)
63-76

Abstract

Antigen detection by sandwich ELISA was evaluaated to predict RT-PCR detection of dengue viral genome in infected culture fluid of Aedes albopictus clone C6/36 cells. Serum specimens collected from dengue patients within five days from onset of fever in two hospitals in Metro Manila, Philippine, were inoculated into C6/36 cells incubated at 28 C.A total of 282 infected culture fluid specimens were harvested and examined by sandwich ELISA and RT-PCR to detect dengue viral antigen and genome respectively. In the sandwich ELISA, the P/N ratio was calculated by dividing optical density (OD) of a given test specimen by the OD of the standard negative specimen. Samples with a P/N ratio > 4.001 were positive for viral genome detection by RT-PCR. The sensitivity and specificity of antigen sandwich ELISA with RT-PCR as the standard were 90.4% and 100% respectively. Although antigen sandwich ELISA is less sensitive than RT-PCR, its usefulness lies in its capability to screen a large number of samples at a minimum cost, especially during an outbreak. Samples that meet a set cutoff value can undergo confirmation of RT-PCR for further epidemiological studies.

 
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