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The page below is a sample from the LabCE course Real-Time PCR. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Reverse Transcriptase PCR (RT-PCR)

PCR can be modified for the amplification of RNA with one additional step prior to the PCR process -- the addition of a retrovirus enzyme called reverse transcriptase. Reverse transcriptase is used to create a copy of DNA using the original RNA specimen. Though there are thermostable polymerases that have reverse transcriptase capabilities, they are not commonly used.

Reverse transcriptase PCR (RT-PCR) is used for the detection of viruses, such as HIV, that have an RNA genome. RT-PCR methods provide early detection of infection, even before the formation of antibodies. Therefore, it is a particularly useful method for HIV and hepatitis C virus (HCV) detection in blood bank nucleic acid testing.

In addition to testing for HIV and HCV, RT-PCR is also used for detection of influenza A virus and other microorganisms where the target is RNA.

RT-PCR is commonly combined with real time PCR.