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Highly specific, quantitative polymerase chain reaction probe for the quantification of human cells in cynomolgus monkeys
- Source :
- Drug metabolism and pharmacokinetics. 36
- Publication Year :
- 2020
-
Abstract
- Quantification of human cells may be performed using quantitative polymerase chain reaction (qPCR). In preclinical studies, the human Alu sequence is widely used as biomarker for human DNA. However, because the Alu gene is shared by primates, its use is limited to non-primate studies. The biodistribution of human cells in primates is also necessary for translational studies. Therefore, we aimed to design a novel, human-specific primer/probe that enables the quantification of human cells in primates and other animal models. A novel primer/probe set was successfully designed based on highly repetitive LINE1 sequences. qPCR efficiency (94.95–99.21%) and linearity of calibration curves (r2 = 0.996–0.999) were confirmed in tissue homogenates of cynomolgus monkey. The lower limit of detection was 10 cells per 15-mg tissue sample, a sensitivity that is equivalent to existing Alu primers/probes. The set was also effective in other animal models such as mice, rabbits, pigs, and common marmosets. To our knowledge, this is the first study describing the successful design of a human-specific qPCR primer/probe for human cell quantification in various animals, including non-human primates, using LINE1 sequence. The excellent selectivity, sensitivity, and versatility of the LINE1 primers/probes make it a promising quantification tool in preclinical biodistribution studies.
- Subjects :
- Male
Biodistribution
Swine
Guinea Pigs
Pharmaceutical Science
Alu element
Tissue sample
Computational biology
Mice, SCID
CD8-Positive T-Lymphocytes
030226 pharmacology & pharmacy
Polymerase Chain Reaction
Cell therapy
03 medical and health sciences
Mice
Young Adult
0302 clinical medicine
Species Specificity
Animals
Humans
Pharmacology (medical)
Tissue Distribution
Gene
030304 developmental biology
Pharmacology
0303 health sciences
Chemistry
Biomarker (cell)
Macaca fascicularis
Real-time polymerase chain reaction
Liver
Rabbits
Primer (molecular biology)
DNA Probes
Subjects
Details
- ISSN :
- 18800920
- Volume :
- 36
- Database :
- OpenAIRE
- Journal :
- Drug metabolism and pharmacokinetics
- Accession number :
- edsair.doi.dedup.....c53f9f4fa4f701970593f09f69fdd0a3