Construction and Identification of a Positive Control Recombinant Plasmid for Respiratory Syncytial Virus Nucleic Acid Detection

Yanlan Wang ； Bo Yu ； Jijian Zhou

《International Core Journal of Engineering》 5卷12期 (2019/11) Pp. 145-148

https://doi.org/10.6919/ICJE.201911_5(12).0022

摘要

Objective: Human respiratory syncytial virus (HRSV) is one of the major viruses of acute respiratory tract disease among infants and young children. Establishing a positive control for HRSV PCR detection is very important for quality control of HRSV PCR detection. Methods: We first designed a specific primer to amplify the HRSV N gene fragment, and inserted it into the pMDTM19-T vector to obtain a recombinant plasmid. The recombinant plasmid was then verified by PCR amplification, Enzyme digestion, sequencing and real-time PCR. Results and Conclusion: Amplification product length and Digestion length of positive control were consistent with the length of the target sequence and a significant amplification curve appeared, suggesting that the positive control can perform good laboratory quality control on PCR detection of HRSV.

關鍵字

Human respiratory syncytial virus ； N gene ； Real-time PCR

參考文獻

Shi T, McAllister D A, O'Brien K L, et al. Global, regional, and national disease burden estimates of acute lower respiratory infections due to respiratory syncytial virus in young children in 2015: A systematic review and modelling study[J]. The Lancet, 2017, 390(10098): 946-958.

Meng J, Hotard A L, Currier M G, et al. The respiratory syncytial virus attachment glycoprotein contribution to infection depends on the specific fusion protein[J]. J Virol, 2015, 90(1): 245.

Zhang Y, Yuan L, Zhang Y, et al. Burden of respiratory syncytial virus infections in china: Systematic review and meta-analysis[J]. J Glob Health, 2015, 5(2): 020417.

Pangesti K N A, Abd El Ghany M, Walsh M G, et al. Molecular epidemiology of respiratory syncytial virus[J]. Rev Med Virol, 2018, 28(2).

Madi N, Chehadeh W, Asadzadeh M, et al. Analysis of genetic variability of respiratory syncytial virus groups a and b in kuwait[J]. Arch Virol, 2018, 163(9): 2405-2413.