Species identification and absolute quantification of biological samples by droplet digital PCR.
- Author:
Wei HU
;
Rong-hua CHEN
;
Chen ZHANG
;
Zhi-yuan AN
;
Bing WANG
;
Yuan PING
- Publication Type:Journal Article
- MeSH:
Animals;
DNA;
DNA Primers/genetics*;
DNA, Mitochondrial/genetics*;
Humans;
NADH Dehydrogenase/genetics*;
Polymerase Chain Reaction/methods*;
RNA, Ribosomal, 16S/genetics*;
Sensitivity and Specificity;
Species Specificity
- From:
Journal of Forensic Medicine
2014;30(5):342-345
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To test droplet digital PCR for species identification and absolute quantification of biological sample.
METHODS:Specific primers and probes for human mtDNA encoding gene ND4 and 16S rRNA were designed, and the species-specificity was assessed on DNA samples derived from human and common animals. To determine the sensitivity and stability of droplet digital PCR for species identification and absolute quantification, gradient dilution series of recombinant plasmid and 16 human DNA samples were analyzed.
RESULTS:Human recombinant plasmid FAM (ND4) could be used in detecting the samples of human. And the results of detecting were consistent with all levels of diluted concentrations. Droplet digital PCR was able to detect low and single copy of target DNA.
CONCLUSION:Droplet digital PCR, with high sensitivity and specificity, is fully amenable for species identification and absolute quantification of biological samples, also it can be applied on routine forensic examination.
