Objective: To evaluate a HBV/HCV/HIV nucleic acids parallel assay kit via PCR and chemiluminescence. Methods: The viral nucleic acids of HBV， HCV and HIV were simultaneously extracted with the homemade magnetic nanoparticles and reagents. The information on the target viral nucleic acids was checked by multiple PCR， electrophoresis, and chemiluminescence. The sensitivity， specificity and stability of the kit were also evaluated. Results: The kit was highly sensitive， specific and stable. The detection limits of the kit were 2.0， 3.7 and 166.6 U/mL for HBV DNA， HCV RNA and HIV RNA， respectively. There was no cross-reactivity with the 10 kinds of viruses, and 2 kinds of bacteria, which might cause interference potentially. The expiry date was about 1 year. Among the 10 422 EIA negative samples of blood donors， 12 were HBV positive and 2 were HCV positive. These results completely coincided with those of other current diagnostic methods of commercial kit applied in blood screening. Conclusion: The kit is fast， accurate， highly sensitive and cost-effective. It can be applicable to the genetic diagnosis and epidemiological survey.