猪瘟病毒C株检测用国家核酸标准物质的研制
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Preparation of National Nucleic Acid Reference Materials of CSFV C Strain for Detection
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    摘要:

    为研制均匀、稳定的猪瘟病毒(CSFV)核酸标准物质,选择CSFV C株接种于PK-15细胞并收获病毒液,然后经过病毒灭活、分装、冻干,最终形成核酸标准物质。用微滴式数字PCR对制备的CSFV标准物质进行均匀性和稳定性评估、标准物质合作定值,在评定不确定度后计算得出最终量值结果,并开展临床试用。结果显示,该标准物质均匀,4 ℃条件下可稳定存放4周,25 ℃可稳定存放1周,-20 ℃可稳定存放11个月,量值结果为(5.1±0.7)×105 copies/mg;临床试用显示,所制备的标准物质与临床样本的互通性良好。本研究制备的CSFV C株国家核酸标准物质均一性良好、稳定、量值准确度高,并通过了全国标准物质管理委员会评定,可用于动物及动物产品质量安全检测,从而有效保障猪瘟防治工作的开展。

    Abstract:

    In order to prepare a kind of uniform and stable nucleic acid reference materials of classical swine fever virus(CSFV),CSFV C strain was selected and inoculated into PK-15 cells to obtain viral fluid,followed by the process of inactivation,subpackage and freeze-drying,then the nucleic acid reference materials were produced. The homogeneity and stability of prepared materials were evaluated by droplet digital PCR,and the value of materials was determined by 8 collaborative laboratories,the quantity value was ultimately calculated after evaluation of uncertainty,then a clinical trial was carried out. The results showed that the material was homogeneous and could be stored stably for four weeks at 4 ℃,for one week at 25 ℃,and for 11 months at -20 ℃,the quantity value was(5.1±0.7)×105 copies/mg;it was found that the materials showed good interactivity with clinical samples in clinical trial. In conclusion,the prepared materials were advantaged by good homogeneity,stability and high accuracy,which had been successfully evaluated by National Administrative Committee for CRM's and could be used in the field of quality and safety detection of animals and animal products,so as to effectively guarantee the prevention and control of CSFV.

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王英超,高晓龙,梅力,高敏,宋彦军,韦海涛,赵启祖,李翠,冯小宇.猪瘟病毒C株检测用国家核酸标准物质的研制[J].《中国动物检疫》编辑部,2020,37(10):114-120.

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  • 在线发布日期: 2020-09-29
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