The Comparison of Efficiency, Sensitivity, and Effect of Inhibitors on Commercial DNA Analysis Kits

Abstract

DNA  fingerprint provides great and useful  information for  human identification  in

criminal investigations  and court cases,  family  relationships and human ancestor study.  To

investigate DNA  fingerprint, a commercial kit based on polymerase chain reaction technique

representing a rapid and convenient use has been chosen to obtain the highest efficiency  and less

time-consuming.  The parameters such as sensitivity, efficiency,  and capability to amplify DNA

under inhibitors were considered for the selection of commercial kit. Hence, the aim of research was to compare these parameters between two commercial kits (Investigator®24 plex Go kit and PowerPlex®   Fusion 6C). The standard DNA was serial two-fold  diluted in the range between

0.25-5.0 ng/µL. In the case of inhibitors, DNA samples were mixed with different kinds of inhibitors (calcium chloride, humic acid, or hematin). The percentage of detected allele, average peak height, heterozygous  peak balance, and inter-locus  balance were calculated. The results found that percentage of detected allele, average peak height, and heterozygous peak balance in low concentration of DNA sample analyzed by Investigator®24 plex Go kit were better than those by PowerPlex®  Fusion 6C kit. However, DNA sample investigated by PowerPlex®  Fusion 6C kit provided low value ranging in acceptance criteria of inter-locus balance. It was found that calcium chloride affected DNA amplification processed by PowerPlex®  Fusion 6C kit, whereas hematin interfered with DNA amplification processed by Investigator®24 plex Go kit. Nevertheless, humic acid did not affect amplification process of both commercial kits. In conclusion, both commercial kits showed efficiency  in DNA fingerprinting. Investigator®24 plex Go kit also gave good typing which can be obtained from a very low amount of DNA.  The research data provides useful information to guide the selection of commercial kit that corresponds with services and specimens in laboratory resulting in maximal accuracy and precision of DNA typing result.