The use of Armored RNA as a multi-purpose internal control for RT-PCR.
Real time reverse transcriptase-PCR (RT-PCR) is now used commonly for the detection of viral pathogens in respiratory samples. However, due to potential inhibition of the RT-PCR or inefficient extraction, this sample type can present significant challenges to accurate patient testing. The goal of this study was to create an internal control to be multiplexed in a real time RT-PCR assay for detecting a viral target in respiratory samples. This report describes an Armored RNA (aRNA) internal control developed originally to be multiplexed in a real time RT-PCR assay for detecting SARS-associated Coronavirus, but can be incorporated into any RT-PCR assay. The internal control primers and probe target a region in the coat protein gene of the E. coli F-specific bacteriophage ms2, which is contained within the aRNA.
Stevenson
Jeffery
J
Associated Regional and University Pathologists, Institute for Clinical and Experimental Pathology, 500 Chipeta Way, Salt Lake City, UT 84108, USA. stevenj@aruplab.com
Hymas
Weston
W
Hillyard
David
D
D017931:DNA Primers / Q000235:genetics; D006801:Humans; D017909:Levivirus / Q000235:genetics; D012313:RNA / Q000737:chemistry*; D012015:Reference Standards; D012141:Respiratory Tract Infections / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D014777:Virus Diseases / Q000175:diagnosis
D017931:DNA Primers; D012313:RNA