期刊
NUCLEIC ACIDS RESEARCH
卷 33, 期 21, 页码 -出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gni181
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资金
- NATIONAL CANCER INSTITUTE [R03CA110136, R01CA109232] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF BIOMEDICAL IMAGING AND BIOENGINEERING [R21EB000983] Funding Source: NIH RePORTER
- NCI NIH HHS [CA109232, R03 CA110136, CA110136, R01 CA109232] Funding Source: Medline
- NIBIB NIH HHS [EB00983, R21 EB000983] Funding Source: Medline
Quantitative real-time PCR has become the method of choice for measuring mRNA transcription. Recently, fast PCR protocols have been developed as a means to increase assay throughput. Yet it is unclear whether more rapid cycling conditions preserve the original assay performance characteristics. We compared 16 primer sets directed against Epstein-Barr virus (EBV) mRNAs using universal and fast PCR cycling conditions. These primers are of clinical relevance, since they can be used to monitor viral oncogene and drug-resistance gene expression in transplant patients and EBV-associated cancers. While none of the primers failed under fast PCR conditions, the fast PCR protocols performed worse than universal cycling conditions. Fast PCR was associated with a loss of sensitivity as well as higher variability, but not with a loss of specificity or with a higher false positive rate.
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