A quantification of human cells using an ERV-3 real time PCR assay.
A novel approach to quantifying human cells using a real time PCR assay was developed. The target sequence used in the assay is a 135 bp segment within the unique 1.7 kb Hind III / Pst I fragment of the ERV-3 envelope gene. ERV-3 is a full-length human endogenous retrovirus present in known copy number in all human cells. The detection range of ERV-3 by real time PCR is from 10(6) to 10(1). The precision described, sensitivity and specificity of the assay indicate that the ERV-3 sequence is an accurate cell quantitation marker. The quantitative ERV-3 assay enables simple, fast, and reproducible detection and quantitation of the cell number. The assay can be used to determine the sample DNA conditions and also it can be used to adjust the quantitative DNA measurements of other target gene assays relative to the number of cell equivalents.
Yuan
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Virus Epidemiology Section, AIDS Vaccine Program, SAIC Frederick, National Cancer Institute, Frederick Cancer Research and Development Center, Frederick, MD 21702, USA. yuan@avpaxp.1ncifcrf.gov
Miley
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Waters
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D000818:Animals; D015415:Biomarkers; D002452:Cell Count / Q000379:methods*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000097:blood; D020077:Endogenous Retroviruses / Q000235:genetics* / Q000302:isolation & purification; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013698:Templates, Genetic; D014407:Tumor Cells, Cultured; D014759:Viral Envelope Proteins / Q000032:analysis / Q000235:genetics
D015415:Biomarkers; D017931:DNA Primers; D004279:DNA, Viral; D014759:Viral Envelope Proteins