猪圆环病毒1型和2型核酸定性标准样品的研制

为了研制猪圆环病毒1型(PCV1)、猪圆环病毒2型(PCV2)核酸定性标准样品,分别将PCV1和PCV2病毒全序列克隆至pMD18-T载体中并进行稀释分装。制备后进行均匀性评估、稳定性评估、定值和临床试用。结果:制备的标准样品的参考值分别为:1.33×10⁴ copies/μL(PCV1)和1.11×10⁴ copies/μL(PCV2);样品均匀;-20℃可稳定保存24个月以上,4℃可稳定2个月以上。经国家标准化管理委员会组织的专家评审,达到了国家标准样品的要求,可作为核酸扩增检测的猪圆环病毒核酸标准样品。

Development of certified reference material for identification of nucleic acid of porcine circovirus type 1 and 2

To develop certified reference materials for identifying nucleic acid of porcine circovirus subtype,the complete genome sequences of PCV1 and PCV2 were cloned into the pMD18-T plasmid,diluted and sub-packed,respectively.The homogeneity and stability of the prepared reference materials were tested.Then,the prepared reference materials were valued using digital PCR and their clinical applicability was also evaluated.The results indicated that the reference values of the reference materials were 1.33×10⁴ copies/μL(PCV1)and 1.11×10⁴ copies/μL(PCV2).The prepared materials would be stable for at least 24 months at-20℃,for more than 2 months at 4℃.The prepared material of PCVs passed the review of Standardization Administration of China and might be used as standard samples for nucleic acid amplification test of nucleic acid of PCV.