DNA Chip For HBV, HCV And HIV-1 Simultaneous Detection

Hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus type 1 (HIV-1) are very dangerous to human and transmitted mainly through transfusion of blood. Since it is difficult to cure the diseases caused by these three kind viruses, detection of these viruses is very important in transfusion and manufacture in blood products. Traditional detection methods are time-consuming and laborious with low detection sensitivity. It is the advantages of DNA chips that are quick, accurate and simultaneous viruses' detection with higher sensitivity.In this research, the mode of DNA chip for virus detection is constructed. Then, the solid phase nested three loci PCR and the enzymatic indicator system are applicated in the three kind viruses' detection in the positive blood samples.The work include:Six plasmids, which contain six kind viruses' specific DNA fragments, were amplified by six loci PCR on a slide on the basis of six loci PCR in a tube.Two plasmids, which contain HBV specific DNA fragment and HSV1 DNA fragment, were amplified by solid phase two loci PCR and detected by enzymatic indicator system on a gene chip that was constructed by primer immobilization and modified with thiol group on chip surface.For building detection technology by DNA chip in clinical, The virus genomes were extracted from the clinical positive samples by one-step nucleic acid extraction. After one-step reverse transcription of the RNA virus genomes, the viruses' specific DNA fragments were amplified by nested three loci PCR, which the amplification system was optimized by uniform design. Finally, the solid phase nested three loci PCR was applied to detect HBV, HCV and HIV-1 in the blood samples and the amplified DNA products on the chip surface were detected by the enzymatic indicator system.