甲型H1N1流感病毒核酸国家参考品的研制

目的研制甲型H1N1流感病毒核酸国家参考品。方法收集甲型H1N1流感病毒及其他病毒培养物,用实时荧光PCR方法逐一进行验证。参考品通过协同标定,确定最低检出限范围,7家单位进行适用性检测。反复冻融观察其稳定性。结果所用的3株病毒经实时荧光PCR验证,特异性良好,经基因芯片进一步验证,均为甲型H1N1流感病毒,且无其他病毒核酸污染,经7家单位检测,符合国家参考品标准要求;反复冻融3次后,稳定性良好。结论建立了第1套甲型H1N1流感病毒核酸国家参考品及相应的质量标准,并已被国家有关部门批准正式使用。

Preparation of National Reference Panel of Influenza A(H1N1)Virus Nucleic Acid

Objective To prepare national reference panel of influenza A(H1N1)virus nucleic acid. Methods The cultures of influenza A (H1N1)virus and other influenza viruses were collected and verified individually by real-time fluorescent PCR. The reference panels were collaboratively calibrated to determine the minimum detection limit, then tested for adaptivity by 7 laboratories, and observed for stability by repeat freezing and thawing. Results Three virus strains showed high specificities by real-time fluorescent PCR and was further verified as influenza A (H1N1)virus free from contamination with other viral nucleic acids by gene chip technique. The test results by 7 laboratories proved that the reference met the requirements for national reference panel. The reference showed good stability after 3 cycles of freezing and thawing. Conclusion The first generation of national reference panel of influenza A (H1N1)virus nucleic acid as well as the relevant quality standard were established and approved by the state departments concerned.