PCR Detection Of Viral DNA/RNA

In clinical molecular diagnostics, nucleic acid amplification tests such as the reverse transcriptase-polymerase chain reaction (RT-PCR) enable the early detection and identification of infectious diseases caused by retroviruses. Utilizing mRNA as its starting template, RT-PCR expeditiously transcribes, amplifies and detects the genetic material of retroviruses in vitro. Nucleic acid tests have been successfully used in the detection of well-known retroviruses including influenza viruses, enteroviruses, coronavirus, Ebola virus, and HIV.

Quantification of RT-PCR products can be generally divided into two categories: end-point RT-PCR and real-time PCR. The use of end-point RT-PCR is preferred for measuring gene expression changes in small number of samples, while real-time RT-PCR has become the standard method for validating gene expression changes on a global scale. Real-time PCR (RT-PCR) fluorescent reporter molecules include dyes that bind to the double-stranded DNA, such as Cyber Green™, or sequence-specific paired FRET probes, such as Tide Fluor™ and Tide Quencher™ dyes.

(Note that AAT Bioquest provides reagents intended for research use only)

RNA dose response with StrandBrite™ Green in a solid black 96-well microplate and measured using a Gemini microplate reader (Molecular Devices).