RNA病毒检测质控物质-大肠杆菌噬菌体MS2标准样品的研制和应用

目的研制大肠杆菌噬菌体MS2标准样品,以其作为RNA病毒检测质控物质,建立RNA病毒检测全过程的质量控制体系。方法利用液体培养法制备大肠杆菌噬菌体MS2标准样品;采用双层琼脂法对标准样品进行稳定性、均匀性测定并定值;将标准样品添加于贝类样品中,应用于贝类诺如病毒实时荧光检测全过程质量控制。结果该标准样品稳定性与均匀性良好,定值为(1.57±0.0288)×10~(11) pfu/m L;保质期为12个月;在4种不同贝类样品中,病毒提取效率在3.70%~7.84%之间,符合国际标准ISO 15216:2-2013的病毒提取效率大于1%的要求。结论该研究制备的大肠杆菌噬菌体MS2标准样品可以对RNA病毒检测全过程进行良好的质量控制,具有良好的应用前景。

Development and application of coliphage MS2 standard material as quality control in RNA virus detection

Objective To investigate coliphage MS2 standard material used as quality control material so as to establish the whole process quality control system of RNA virus detection in which the standard material was used as quality control. Methods Standard materials of coliphage MS2 were developed by liquid culture method, and the stability, uniformity and valuing of the standard materials were also determined by double agar plate method. The standard material was applied in real-time fluorescent RT-PCR for detection of norovirus in shellfish to guarantee the quality of whole detection process. Results The standard material had good stability and uniformity and was valued as (1.57±0.0288)×1011 pfu/mL, and the shelf life was determined as 12 months. The extraction efficiencies of virus were between 3.70% and 7.84% in four kinds of shellfish samples, which were according with ISO 15216:2-2013 (virus extraction efficiency should be more than 1%). Conclusion The coliphage MS2 standard material is good at the whole process of quality control in RNA virus detection, which has a promising prospect.