HPV16与HPV18假病毒核酸标准物质定值研究

The Research of Quantification for HPV16 and HPV18 Pseudovirus Nucleic Acid Reference Materials

以分别包含HPV16、HPV18 L1基因序列的假病毒为候选材料,通过数字PCR定量方法对L1基因的拷贝数浓度进行检测,同时验证了标准物质的均匀性和稳定性,研制了HPV16与HPV18假病毒核酸标准物质(NIM-RM5213和NIM-RM5214)。2种标准物质的拷贝数标准值及扩展不确定度(k=2)分别为:(1.52±0.13)×10^(3)copies/μL,(1.29±0.12)×10^(3)copies/μL。利用多家检测平台对2种标准物质进行定值验证,定值结果的相对标准偏差均在5%以内。该标准物质能够为HPV检测全过程的质量控制提供参考,有助于提升HPV核酸检测过程的准确性与有效性,具有广泛的应用前景。

Pseudoviruses containing L1 gene sequences of HPV16 and HPV18 were selected as reference materials(NIM-RM5213 and NIM-RM5214).Employing the digital PCR quantification method,we detected the copy number concentration of L1 gene,as well as the verified the uniformity and stability.The reference values with expanded uncertainty of HPV16 and HPV18 reference material were(1.52±0.13)×10^(3) copies/μL,(1.29±0.12)×10^(3) copies/μL,respectively.Multiple testing platforms were recruited to validate the values of the two reference materials,and the relative standard deviations of their values were less than 5%.These reference materials could be applied to quality control of the whole process of HPV detection.They help to improve the accuracy and effectiveness of the HPV nucleic acid detection and have a broad application prospect.Using pseudoviruses containing HPV16 and HPV18 L1 gene sequences as candidate materials,the copy number concentration of L1 gene was detected by digital PCR quantitative method,and the homogeneity and stability of the reference materials were verified.HPV16 and HPV18 pseudovirus nucleic acid reference materials(NIM-RM5213 and NIM-RM5214)were developed.The standard value of copy number and expanded uncertainty(k=2)of the two reference materials are(1.52±0.13)×10^(3) copies/μL,(1.29±0.12)×10^(3) copies/μL。The calibration results of the two reference materials were verified by multiple testing platforms,and the relative standard deviation of the calibration results was within 5%.The mentioned standard material can provide reference for the quality control of the whole process of HPV detection,help to improve the accuracy and effectiveness of HPV nucleic acid detection process,and has broad application prospects.