Optimization Of Real-Time Reverse Transcriptase Polymerase Chain Reaction For Detection Of Dengue Virus

Abstract Objective Rapid and accurate laboratory confirmatory is very essential for control measures of dengue virus infections. However, many cases of dengue virus infections in most of the hospitals remain undiagnosed due to presence of other febrile illnesses with overlapping symptoms and lack of specificity in most of laboratory diagnostic methods. This study was set to optimize conditions for real time reverse transcriptase polymerase chain reaction (RT-PCR) for detection of dengue virus by using rapid and simple nucleic acid extraction method.Results The real time RT-PCR technique was successfully optimized using simple and rapid method for purification of nucleic acid, ‘boom method’. The technique works better when performed in a two-step procedure and can works well with all range of real time PCR machines. The optimized real time RT-PCR used in the present study is a valuable and reliable technique for routine diagnosis of dengue. Further investigation on the cost effectiveness in adopting this technique for routine screening and monitoring of the dengue infection should be done.