一步法mRNA-PCR荧光复合扩增检测体系在法医案件中的验证OA

To evaluate the application potential of the one-step mRNA-PCR fluorescence multiplex amplification detection technology for identifying body fluid stains in case samples,a total of 89 samples suspected to be blood,saliva,semen,and vaginal secretions were collected.These samples were analyzed collaboratively with two forensic DNA laboratories lacking prior RNA operation experience.The laboratories performed total RNA extraction,one-step RT-PCR amplification,and capillary electrophoresis detection using different experimental instruments.The results demonstrated that three housekeeping genes(GAPDH,ACTB,and PRL19)were successfully detected in all 89 case samples.Additionally,specific markers for each body fluid were identified:HBB and HBA for peripheral blood;MMP7 and MMP10 for menstrual blood;STATH and HTN3 for saliva;PRM2 and SEMG1 for semen;and CYP2B7P1 and HBD1 for vaginal fluid.Notably,each participating DNA laboratory was able to independently complete the detection of RNA markers in the case samples,and parallel testing of the same samples across different laboratories yielded consistent results.This study provides robust evidence supporting the application of the one-step mRNA-PCR fluorescence multiplex amplification detection system for the source identification of body fluid stains in forensic case samples.