Optimization of Real-time Reverse Transcriptase Polymerase Chain Reaction for Detection of Dengue Virus

Main Article Content

Kaunara A. Azizi
Arnold J. Ndaro
Athanasia Maro
Adonira Saro
Reginald A. Kavishe

Abstract

Aims: 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.

Methodology: One step and two step real time RT-PCR were evaluated in different PCR thermocyclers. Extraction of viral RNA was done by using a simple boom 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.

Keywords:
Dengue virus, RT-PCR technique, boom method, RNA extraction.

Article Details

How to Cite
Azizi, K. A., Ndaro, A. J., Maro, A., Saro, A., & Kavishe, R. A. (2020). Optimization of Real-time Reverse Transcriptase Polymerase Chain Reaction for Detection of Dengue Virus. Journal of Advances in Microbiology, 20(8), 1-7. https://doi.org/10.9734/jamb/2020/v20i830269
Section
Original Research Article

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