Abstract：
:Quantitative PCR assays are now the standard method for viral diagnostics. These assays must be specific, as well as sensitive, to detect the potentially low starting copy number of viral genomic material. We describe a new technique, polymerase chain displacement reaction (PCDR), which uses multiple nested primers in a rapid, capped, one-tube reaction that increases the sensitivity of normal quantitative PCR (qPCR) assays. Sensitivity was increased by approximately 10-fold in a proof-of-principle test on dengue virus sequence. In PCDR, when extension occurs from the outer primer, it displaces the extension strand produced from the inner primer by utilizing a polymerase that has strand displacement activity. This allows a greater than 2-fold increase of amplification product for each amplification cycle and therefore increased sensitivity and speed over conventional PCR. Increased sensitivity in PCDR would be useful in nucleic acid detection for viral diagnostics.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Harris CL,Sanchez-Vargas IJ,Olson KE,Alphey L,Fu Gdoi
10.2144/000113951subject
Has Abstractpub_date
2013-02-01 00:00:00pages
93-7issue
2eissn
0736-6205issn
1940-9818pii
000113951journal_volume
54pub_type
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