Study of Four STR Loci Fluorescent Multiplex PCR Amplification System and Its Application in Forensic Genetics
|Keywords||Forensic medicine DNA type Short Tandem Repeat PCR Fluorescent Multiplex amplification ABI-310 Population genetics|
Objective The purpose of this project is to establish the fluorescent MultiplexSTR system of D11S4951, D7S3062, GATA168F06 and D10S1426. Andvalidate the forensic application of this multiplex system under the guidelinesof TMGDAM. Methods Select D11S4951, D7S3062, GATA168F06 andD10S1426, with good polymorphisms and high Discriminate Probabilityfrom the loci that have been undertaken the population genetic research byour laboratory. And then begin the research of this fluorescent multiplexsystem using the traditional multiplex methods. Using designed primer set,four STR loci D11S4951, D7S3062, GATA168F06 and D10S1426, wereamplified and the products of the multiplex PCR were genotyped by ABI-310 Genetic Analyzer. The multiplex STR system was evaluated the sensitivity,accuracy and different PCR outcome under different cycle times, etc.Validation study of Forensic Science, according to the guidelines ofTMGDAM, included species specificity, tissular identity, repeatability andperformance of typing forensic samples. Result Successfully establish thefluorescent multiplex system of D11S4951, D7S3062, GATA168F06 andD10S1426. Moreover, it is easy to use. We can get the satisfied analytic resultonly using the home-made Taq polymerase and doesn’t need the hot start ofPCR. And compared with the commercial kit it is low cost and higher-performance. Validation study of Forensic Science shows that the samplesensitivity of this system is 0.25ng template DNA, and the four loci’s speciesspecificity and tissular identity is good. This system can make successfullygenotype mixed samples and the samples which come from different basematerial. And the utilization of fifteen forensic cases proves the application ofthis system in forensic individual identification and paternity test.Conclusion The multiplex system of D11S4951, D7S3062, GATA168F06and D10S1426 can be successfully used in the ABI-310 detection platformand can get the reliable genotype. The forensic genetics research implies thatit is an easily-regulated, low cost multiplex system with stable outcome, highsensitivity, accuracy, tissular identity and good species specificity. It canperfect detect the timeworn samples, mixed samples and samples come fromdifferent base material without cross reaction of common animals andmicroorganism. Based on the above advantages, it could be recommended tothe forensic genetics practice.