Abstract:In order to develop plasmid nucleic acid reference material of porcine reproductive and respiratory syndrome virus (PRRSV), a digital PCR method was established, and several laboratories were combined to conduct cooperative evaluation of the reference material by using the digital PCR method. The factors affecting the accuracy of plasmid evaluation in the process of cooperative evaluation were discussed, and the uncertainty of the reference material was evaluated. The two reference materials have been certified with codes GBW(E)091038 and GBW(E)091039, which can provide quantitative standards for detection methods and can be used in method evaluation, product quality control and many other aspects.
牛春艳,张永卓,杨佳怡,董莲华,傅博强,王晶. 基于数字PCR的质粒核酸标准物质合作定值研究[J]. 计量学报, 2021, 42(11): 1522-1527.
NIU Chun-yan,ZHANG Yong-zhuo,YANG Jia-yi,DONG Lian-hua,FU Bo-qiang,WANG Jing. Resrach on the Collaborative Value Assignment of Plasmid Nucleic Acid Refernce Materials Based on Digital PCR. Acta Metrologica Sinica, 2021, 42(11): 1522-1527.
[1]JJF 1001-2011通用计量术语及定义[S].
[2]陈仕龙, 陈少莺, 江斌, 等. 3种鉴别诊断PRRSV美洲型经典毒株和变异株的方法比较[J]. 中国动物传染病学报, 2011, 19(4): 71-75.
Chen S L, Chen S Y, Jiang B, et al. Comparative study of three diagnostic methods for differential detection of classical PRRSV and PRRSV variant [J]. Chinese Journal of Animal Infectious Diseases, 2011, 19(4): 71-75.
[3]Tian H, Wu, J Y, Shang Y J, et al. The development of a rapid SYBR one step real-time RT-PCR for detection of Porcine Reproductive and Respiratory Syndrome Virus [J]. Virol J, 2010, 7: 90.
[4]Wernike K, Hoffmann B, Dauber M, et al. Detection and typing of highly pathogenic porcine reproductive and respiratory syndrome virus by multiplex real-time rt-PCR [J]. PLoS One, 2012, 7(6): 38251.
[5]Yang Q, Xi L, Chen X X, et al. The development of a sensitive droplet digital PCR for quantitative detection of porcine reproductive and respiratory syndrome virus [J]. Int J Biol Macromol, 2017, 104(Pt A): 1223-1228.
[6]Vogelstein B, Kinzler K W. Digital PCR [J]. Proc Natl Acad Sci, 1999, 96(16): 9236-9241.
[7]Sanders R, Huggett J F, Bushell C A, et al. Evaluation of digital PCR for absolute DNA quantification [J]. Analytical Chemistry, 2011, 83(17): 6474-6484.
[8]White R A, Blainey P C, Fan H C, et al. Digital PCR provides sensitive and absolute calibration for high throughput sequencing[J]. BMC Genomics 10, 2009: 116.
[9]Pinheiro L B, Coleman V A, Hindson C M, et al. Evaluation of a droplet digital polymerase chain reaction format for DNA copy number quantification[J]. Analytical Chemistry, 2012, 84(2): 1003-1011.
[10]董莲华, 李亮, 王晶, 等. 转基因玉米pUC57-BT11质粒标准分子的构建与适用性研究[J]. 计量学报, 2011, 32(6): 570-574.
Dong L H, Li L, Wang J, et al. Plasmid pUC57-BT11 construction and application of genetic modified maize line BT11[J]. Acta Metrologica Sinica, 2011, 32(6): 570-574.
[11]牛春艳, 杨佳怡, 王晶, 等. 逆转录过程对猪繁殖与呼吸综合症病毒定量检测的影响[J]. 中国动物传染病学报, 2020, 28(2): 69-74.
Niu C Y, Yang J Y, Wang J, et al. Effect of reverse transcription process on quantitative detection of porcine reproductive and respiratory syndrome virus[J]. Chinese Journal of Animal Infectious Diseases, 2020, 28(2): 69-74.
[12]Dong L H, Meng Y, Sui Z W, et al. Comparison of four digital PCR platforms for accurate quantification of DNA copy number of a certified plasmid DNA reference material[J]. Sci Rep, 2015, 5: 13174.
[13]刘思章, 隋志伟, 张韬, 等. 噬菌体X174标准物质定值方法研究[J]. 计量学报, 2019, 40 (3): 511-516.
Liu S Z, Sui Z W, Zhang T, et al. Study on Measurement for Quantitative Detection of Escherichia Coli [J]. Acta Metrologica Sinica, 2019, 40 (3): 511-516.
[14]陈敏璠, 傅博强, 林婧, 等. 阴沟肠杆菌定量标准物质的研制[J]. 计量学报, 2017, 38 (5): 527-531.
Chen M F, Fu B Q, Lin J, et al. Development of Quantitative Reference Material of Enterobacter Cloacae [J]. Acta Metrologica Sinica, 2017, 38 (5): 527-531.
[15]JJF 1343-2012 标准物质定值的的通用原则及统计学原理[S].