Multiplex amplification system for identifying whether body fluid to be detected is non-blood, menstrual blood or peripheral blood and primer combination used by multiplex amplification system
A compound amplification system and primer combination technology, applied in the field of forensic technology, can solve the problems of time-consuming, laborious, false negative, false positive, etc.
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Embodiment 1
[0090] Embodiment 1, the preparation of the compound amplification system based on 6 genes
[0091] 1. Screening of specifically expressed genes
[0092] The inventors of the present invention obtained a large number of candidate genes specifically expressed in peripheral blood, menstrual blood, saliva, semen, and vaginal secretions by consulting the literature, comprehensively considered the sequence structure of each gene, and finally screened and obtained 4 specifically expressed genes (respectively) HBA gene, HBB gene, MMP7 gene, MMP10 gene) and two housekeeping genes (GAPDH gene and ACTB gene respectively).
[0093] 2. Preparation of multiple amplification system based on 6 genes
[0094] According to the nucleotide sequence of each gene and the length range of the amplified fragment (between 100bp-300bp), different amplified fragment lengths were reasonably selected to distinguish each gene, and then a multiple amplification system based on 6 genes was prepared. Specif...
Embodiment 2
[0143] Example 2. Application of multiple amplification system based on 6 genes in identifying whether the body fluid to be tested is non-blood, menstrual blood or peripheral blood
[0144] 1. Sample collection
[0145] The samples to be tested were 80 samples provided by 80 volunteers (aged 18-35) from Henan. 80 volunteers were unrelated individuals, and all of them gave informed consent.
[0146] The 20 samples numbered PB1-PB20 are all peripheral blood samples. Peripheral blood samples were obtained from venous blood drawn from the arms of some volunteers and stored at -80°C.
[0147] The 20 samples numbered MB1-MB20 are all menstrual blood samples. The 20 menstrual blood samples were collected by some volunteers with tampons 4 days before the menstrual cycle, dried at room temperature for 1 day, and stored at -80°C.
[0148] The 20 samples numbered SA1-SA20 are all saliva samples. The 20 saliva samples were obtained by collecting saliva from some volunteers in sterile...
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