Preparation and application of pseudovirus armor RNA standard quality control for Sendai virus
10.13200/j.cnki.cjb.004293
- VernacularTitle:仙台病毒假病毒装甲RNA标准质控品的制备及其应用
- Author:
QI Xinyao
- Publication Type:Journal Article
- Keywords:
Sendai virus(SeV);
Ms2 bacteriophage;
Pseudovirus;
Armor RNA;
Standard quality control
- From:
Chinese Journal of Biologicals
2024;37(9):1085-1089+1095
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare a pseudovirus armor RNA standard control containing Sendai virus(SeV)RNA by Ms2 bacteriophage in order to use it in the diagnosis of SeV infection and the molecular biological safety detection of the virus.Methods The L gene fragment of SeV was inserted into plasmid pET-28b-Ms2 to construct recombinant plasmid pET-28b-Ms2-L(SeV),which was transformed into competent E.coli BL21(DE3)and induced by IPTG to express the recombinant protein,SeV pseudovirus armor RNA standard quality control. The morphology of the standard quality control was observed under electron microscope,and 10% SDS-PAGE analysis,anti-DNaseⅠ and anti-RNsaeA test,as well as stability analysis were performed. SeV,mouse hepatitis virus(MHV),pneumonia virus of mice(PVM),Hantaan virus(HV),reovirus type Ⅲ（ReoⅢ）and minute virus of mice(MVM)were detected by recombinase polymerase amplification(RPA)to verify the effect of the standard quality control. Results The standard quality control showed the structure of Ms2 bacteriophage pseudoviral particles under electron microscope,with a relative molecular mass of about 14 000,which effectively resisted the degradation of DNaseⅠand RNsaeA. The standard quality control was stored at-80 ℃ for 6 months,-20 ℃ for 6 months and 28 ℃ for 5,10 and 15 days,and still had good stability. RPA specific amplification curve was seen only in SeV,while no amplification curve was observed in other viruses. Conclusion The standard quality control of SeV pseudovirus armor RNA constructed in this study has good stability and specificity,and can be used as a positive control for various molecular biological safety detections of SeV.
