核糖核酸标准物质研制与应用进展

doi:10.13560/j.cnki.biotech.bull.1985.2023-1202

摘要/Abstract

摘要：

核糖核酸（RNA）标准物质是保证核酸体外诊断数据准确一致的重要实物标准，广泛用于病毒类疾病的检测、仪器校准、试剂盒评价及能力验证等工作中，其目的是保证测量过程和测量结果的准确一致。自1997年，WHO研制出了首个针对核酸扩增技术的丙型肝炎核酸国际标准物质，使全球不同实验室、不同方法检测的结果有了可比性。截至2023年，中国在国家标准物质资源共享平台发布的RNA有证标准物质有近百种，对于RNA标准物质的研究与应用，日益受到世界各国计量机构、相关国际学术组织与企业的重视，尤其是新型冠状病毒疫情暴发后，RNA标准物质成为了生物标准物质的研制热点。随着标准物质研制技术的不断发展，标准物质的量值准确性也有所提升，为测量结果的准确一致提供保障。通过对当前RNA标准物质的制备方法、分装、定值技术、不确定度评定及应用领域进行综述，为RNA标准物质的研制工作以及未来在临床医学、农业生产、动物医学、食品安全等领域的应用提供参考。

关键词: 核糖核酸, 标准物质, 标准物质定值, 稳定性, 均匀性, 不确定度评定

Abstract:

RNA reference materials is an important physical standard to ensure the accuracy and consistency of nucleic acid in vitro diagnostic data. It is widely used in the detection of viral diseases. It can also be used in calibration instruments, evaluation kits and proficiency testing, which purpose is to ensure the accuracy and consistency of the measurement process and measurement results. Since 1997, WHO has developed the first hepatitis C nucleic acid international reference material for nucleic acid amplification technology, which makes the results of different laboratories and different methods in the world comparable. By 2023, there are nearly 100 kinds of RNA certified reference materials released by China on the National Sharing Platform for Reference Materials. The research and application of RNA reference materials have been paid more and more attention by metrology institutions, relevant international academic organizations and enterprises all over the world. Especially after the outbreak of the novel coronavirus epidemic, the development of RNA reference materials has become a hot spot in the development of biological reference materials. With the continuous development of the reference materials development technology, the accuracy of the reference materials has also been improved, which provides a guarantee for the accuracy and consistency of the measurement results. The preparation methods, sub-package, fixed value technology, uncertainty evaluation and application fields of RNA reference materials are briefly reviewed, which provides a reference for the development of RNA reference materials and their future applications in various fields.

Key words: RNA, reference material, characterization of reference materials, stability, homogeneity, evaluation of uncertainty

王丹, 赵文江, 齐鑫, 李欣然, 李凯, 樊丽, 李亮. 核糖核酸标准物质研制与应用进展[J]. 生物技术通报, 2024, 40(6): 105-113.

WANG Dan, ZHAO Wen-jiang, QI Xin, LI Xin-ran, LI Kai, FAN Li, LI Liang. Progress in the Development and Application of RNA Reference Materials[J]. Biotechnology Bulletin, 2024, 40(6): 105-113.

图/表 2

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标准物质名称Name of reference standard	定值结果Quantification result	不确定度Uncertainty/%	参考文献Reference
丙型肝炎病毒脱氧核糖酸血清标准物质	1.29×10⁵ IU/mL	18.6	[8]
猪繁殖与呼吸综合征病毒欧洲株（PRRSV LV）核酸标准物质	1.83×10⁴ copies/μL	12	[12]
新城疫病毒（NDV Lasota株）核酸标准物质	1.7×10⁵ copies/mg	17.6	[27]
新型冠状病毒体外转录RNA标准物质（新冠） ORF1ab基因	7.70×10⁵ copies/μL	16.7	[28]
新型冠状病毒体外转录RNA标准物质（新冠） E基因	4.30×10⁵ copies/μL	15	[28]
新型冠状病毒体外转录RNA标准物质（新冠） N基因	6.10×10⁵ copies/μL	23	[28]
HPV16 E6/E7 RNA假病毒标准物质	1.10×10³ copies/μL	17	[29]
猪繁殖与呼吸综合征病毒美洲变异株（PRRSV JXA1）核酸标准物质	1.13×10⁴ copies/μL	16	[30]

标准物质名称Name of reference standard	定值结果Quantification result	不确定度Uncertainty/%	参考文献Reference
丙型肝炎病毒脱氧核糖酸血清标准物质	1.29×10⁵ IU/mL	18.6	[8]
猪繁殖与呼吸综合征病毒欧洲株（PRRSV LV）核酸标准物质	1.83×10⁴ copies/μL	12	[12]
新城疫病毒（NDV Lasota株）核酸标准物质	1.7×10⁵ copies/mg	17.6	[27]
新型冠状病毒体外转录RNA标准物质（新冠） ORF1ab基因	7.70×10⁵ copies/μL	16.7	[28]
新型冠状病毒体外转录RNA标准物质（新冠） E基因	4.30×10⁵ copies/μL	15	[28]
新型冠状病毒体外转录RNA标准物质（新冠） N基因	6.10×10⁵ copies/μL	23	[28]
HPV16 E6/E7 RNA假病毒标准物质	1.10×10³ copies/μL	17	[29]
猪繁殖与呼吸综合征病毒美洲变异株（PRRSV JXA1）核酸标准物质	1.13×10⁴ copies/μL	16	[30]