title: Antigen sandwich ELISA predicts RT-PCR detection of dengue virus genome in infected culture fluids of Aedes albopictus C6/36 cells.
abstract: Antigen detection by sandwich ELISA was evaluated 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 5 days from onset of fever in 2 hospitals in Metro Manila, Philippines, were inoculated into C6/36 cells, and incubated at 28 degrees 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 > or = 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 by RT-PCR for further epidemiological studies.
authors.affiliation: Institute of Biology, College of Science, University of the Philippines, Diliman Quezon City.
authors.forename: Corazon C
authors.initials: CC
authors.lastname: Buerano
authors.forename: Filipinas F
authors.initials: FF
authors.lastname: Natividad
authors.forename: Rodolfo C
authors.initials: RC
authors.lastname: Contreras
authors.forename: Ima Nurisa
authors.initials: IN
authors.lastname: Ibrahim
authors.forename: Marlou Noel M
authors.initials: MN
authors.lastname: Mangada
authors.forename: Futoshi
authors.initials: F
authors.lastname: Hasebe
authors.forename: Shingo
authors.initials: S
authors.lastname: Inoue
authors.forename: Ronald R
authors.initials: RR
authors.lastname: Matias
authors.forename: Akira
authors.initials: A
authors.lastname: Igarashi
mesh_terms: D000330:Aedes / Q000166:cytology / Q000821:virology*; D000818:Animals; D000956:Antigens, Viral / Q000032:analysis*; D003716:Dengue Virus / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D016679:Genome, Viral; D006801:Humans; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D000956:Antigens, Viral