VJPM
Page: 49
Volume 28, Issue 10(209) 2018
Preparation of Armored Dengue virus RNA particle for quality control in RT-PCR assays
Author: Ha Thi Phuong Mai, Tran Thi Hai Au, Do Thi Quynh Nga, Vu Thi Kim Lien, Nguyen Thi Thu Thuy and Le Thi Quynh Mai
Summary:
Armored RNA (AR) technology is a mean for producing nuclease-resistant RNA positive controls or standards for RT-PCR assays. AR is a complex of MS2 bacteriophage coat protein and RNA fragment of a target gene produced in Escherichia coli. The advantages of phage control consist of the ability to protect the control RNA contained in their capsid from degradation by ubiquitous ribonucleases, the safety due to their non-pathogenicity to humans and the stability in room temperature (25°C to 37°C). In this study, we successfully produced MS2-phage like particles carrying internal control prepared for monitoring procedures of Reverse transcription (RT)-PCR for serotyping of Dengue virus infection. AR-DEN was completely resistant to DNase and RNase under conditions in which naked DNA and RNA were both degraded rapidly. Moreover, AR shows high stability at room temperature (25°C to 37°C) for 24 hours, 48 hours, 72 hours and 7 days; 3 and 6 months at -300C. In conclusion, AR-DEN could be used as a positive control or standard in RT-PCR assays for the detection and serotyping of Dengue virus
Keywords:
Armored RNA technology; quality control; RT-PCR; dengue virus
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6. Ha Thi Phuong Mai.pdf
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