Abstract: ObjectiveTo prepare a safe and stable positive control for real-time RT-PCR (rRT-PCR) detection of Xinjiang hemorrhagic fever virus.MethodsThe nucleic acid sequence containing a rRT-PCR amplification target of Xinjiang hemorrhagic fever virus was synthesized and inserted into a lentiviral vector, the recombinant lentiviral vector and the lentivirus-packaging vector were co-transfected into 293T cells. After 48 h of transfection, the rRT-PCR assay was used for detection of the pseudoviral particles packaging and production. Then the pseudoviral particles were collected and treated with nucleic acid protective agent to prepare a positive control, and then the determination of stability was conducted.ResultsAfter the recombinant lentiviral vector and lentivirus-packaging vector were co-transfected into 293T cells, the supernatant was harvested and detected by rRT-PCR. Ten-fold serial dilutions from 10-1 to 10-4 of supernatant were detected, the result showed that the Ct value ranged from 13 to 33, indicating that the concentration of pseudoviral particles in the supernatant was high. The 1 000-fold diluted supernatant was used for preparing a positive control. The stability test was conducted after the positive control was placed in the incubator at 37 ℃ for 0, 3, 7, 15, 21, and 30 days. Ct value of the rRT-PCR detection was 25 to 28 approximately, demonstrating that the positive control possessed high heat stability.ConclusionThe pseudoviral particles produced by using lentiviral packaging technology in this study can be used for an ideal positive control for rRT-PCR detection of Xinjiang hemorrhagic fever virus.