Editing Reverse transcription polymerase chain reaction (section)

==History==

Since its introduction in 1977, [[Northern blot]] has been used extensively for RNA quantification despite its shortcomings: (a) time-consuming technique, (b) requires a large quantity of RNA for detection, and (c) quantitatively inaccurate in the low abundance of RNA content.<ref name="pmid414220">{{cite journal |vauthors=Alwine JC, Kemp DJ, Stark GR |title=Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes |journal=Proc. Natl. Acad. Sci. U.S.A. |volume=74 |issue=12 |pages=5350–4 |date=December 1977 |pmid=414220 |pmc=431715 |doi= 10.1073/pnas.74.12.5350|bibcode=1977PNAS...74.5350A |doi-access=free }}</ref><ref name="pmid19131955">{{cite journal |vauthors=Streit S, Michalski CW, Erkan M, Kleeff J, Friess H |title=Northern blot analysis for detection and quantification of RNA in pancreatic cancer cells and tissues |journal=Nat Protoc |volume=4 |issue=1 |pages=37–43 |year=2009 |pmid=19131955 |doi=10.1038/nprot.2008.216 |s2cid=24980302 }}</ref> However, since [[Polymerase chain reaction|PCR]] was invented by [[Kary Mullis]] in 1983, RT PCR has since displaced Northern blot as the method of choice for RNA detection and quantification.<ref>{{cite journal |author=Bustin SA |title=Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays |journal=J. Mol. Endocrinol. |volume=25 |issue=2 |pages=169–93 |date=October 2000 |pmid=11013345 |doi= 10.1677/jme.0.0250169|doi-access=free }}</ref>

RT-PCR has risen to become the benchmark technology for the detection and/or comparison of RNA levels for several reasons: (a) it does not require post PCR processing, (b) a wide range (>10<sup>7</sup>-fold) of RNA abundance can be measured, and (c) it provides insight into both qualitative and quantitative data.<ref name="pmid15956331"/> Due to its simplicity, specificity and sensitivity, RT-PCR is used in a wide range of [[Applications of PCR|applications]] from experiments as simple as quantification of [[yeast|yeast cells]] in wine to more complex uses as diagnostic tools for detecting infectious agents such as the [[avian flu]] [[virus]] and [[SARS-CoV-2]].<ref name="pmid17088381">{{cite journal |vauthors=Hierro N, Esteve-Zarzoso B, González A, Mas A, Guillamón JM |title=Real-time quantitative PCR (QPCR) and reverse transcription-QPCR for detection and enumeration of total yeasts in wine |journal=Appl. Environ. Microbiol. |volume=72 |issue=11 |pages=7148–55 |date=November 2006 |pmid=17088381 |pmc=1636171 |doi=10.1128/AEM.00388-06 |bibcode=2006ApEnM..72.7148H }}</ref><ref name="pmid19627372">{{cite journal  |vauthors=Slomka MJ, Pavlidis T, Coward VJ, etal |title=Validated RealTime reverse transcriptase PCR methods for the diagnosis and pathotyping of Eurasian H7 avian influenza viruses |journal=Influenza and Other Respiratory Viruses |volume=3 |issue=4 |pages=151–64 |date=July 2009 |pmid=19627372 |pmc=4634683 |doi=10.1111/j.1750-2659.2009.00083.x }}</ref><ref>Mission summary: WHO Field Visit to Wuhan, China 20–21 January 2020: https://www.who.int/china/news/detail/22-01-2020-field-visit-wuhan-china-jan-2020</ref>