Detection and typing of dengue virus using polymerase chain reaction and microwell plate hybridization / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To establish a specific, sensitive and practicable method for detection and typing of dengue virus.</p><p><b>METHODS</b>Based on the genomic sequence analysis of dengue virus types 1-4, 4 pairs of primers were designed. The specific capture probes of dengue virus types 1-4 were amplified using RT-PCR, cloned and sequenced before using them for precoating the microwell plate. The samples were amplified using biotin-labeled forward primer and reverse primer, and microwell plate hybridization was carried out for detection and typing of dengue virus types 1-4.</p><p><b>RESULTS</b>The absorbance of the positive samples were higher than 0.5, while the average absorbance of the negative samples was lower than 0.1, with the S/N higher than 10.</p><p><b>CONCLUSION</b>The method of PCR-ELISA we established for early detection and typing of all 4 dengue viruses seretypes.</p>