Back to Search
Start Over
Automated type specific ELISA probe detection of amplified NS3 gene products of dengue viruses
- Source :
- Journal of Clinical Pathology. 50:346-349
- Publication Year :
- 1997
- Publisher :
- BMJ, 1997.
-
Abstract
- AIM: To apply an automated system of nucleic acid hybridisation coupled with the enzyme linked immunosorbent assay (ELISA) for the type specific detection of amplification products of dengue viruses. METHODS: Non-structural 3 (NS3) gene targets of reference strains of all four dengue and other flaviviruses, as well as dengue patient viraemic sera, were subjected to reverse transcription and polymerase chain reaction using consensus and dengue type specific primers and digoxigenin-11-dUTP label incorporation. The amplification products were detected by biotinylated type specific primers which served as ELISA capture probes bound to streptavidin coated tubes. RESULTS: Significantly high spectrophotometric absorbance readings were obtained by hybridisation of the consensus and seminested amplification products of all four dengue viruses with their respective capture probes. In contrast, extremely low absorbances were observed for consensus products of Japanese encephalitis, yellow fever, and Kunjin viruses, which served as negative controls. These ELISA data correlated well with agarose gel electrophoresis of dengue type specific amplified products of diagnostic sizes. CONCLUSIONS: The combination of in vitro amplification and antibody based detection offers rapid, type specific, high throughput, and gel-free detection of amplified products of dengue viruses.
- Subjects :
- Genes, Viral
Enzyme-Linked Immunosorbent Assay
Viral Nonstructural Proteins
Polymerase Chain Reaction
Sensitivity and Specificity
Pathology and Forensic Medicine
law.invention
Dengue fever
Dengue
Flaviviridae
Nucleic acid thermodynamics
law
medicine
Humans
Polymerase chain reaction
Retrospective Studies
biology
Flavivirus
Serine Endopeptidases
Nucleic Acid Hybridization
General Medicine
biology.organism_classification
medicine.disease
Virology
Molecular biology
Reverse transcriptase
Biotinylation
Agarose gel electrophoresis
RNA Helicases
Research Article
Subjects
Details
- ISSN :
- 00219746
- Volume :
- 50
- Database :
- OpenAIRE
- Journal :
- Journal of Clinical Pathology
- Accession number :
- edsair.doi.dedup.....80d3bf40f671b2bde38af252d46bec0a
- Full Text :
- https://doi.org/10.1136/jcp.50.4.346