论文部分内容阅读
目的建立果蔬生产用水中甲型肝炎病毒(HAV)的检测方法和质量控制标准体系。方法在水样中添加2.5×10~9pfu/m L的质控物质大肠杆菌噬菌体MS2100μL,采用正电荷滤膜法捕获病毒,并根据大肠杆菌噬菌体MS2的标准曲线计算出病毒回收效率,实时荧光RT-PCR检测水样中甲肝病毒,并建立质量控制标准体系。结果对6个水质样品同时加入HAV和大肠杆菌噬菌体MS2并进行病毒回收率的比较,HAV回收率为1.24%~32.68%,MS2回收率为1.64%~14.24%,达到了ISO/TS 15216-2-2013对病毒回收率>1%的要求。同时对30个实际水质样品进行HAV检测,1个样品检出HAV,质控物质大肠杆菌噬菌体MS2提取回收率为1.24%~24.19%,标准偏差为0.0612。结论所建立的水样中病毒检测质量控制体系保证了检测结果质量。
Objective To establish a method and quality control system for detection of hepatitis A virus (HAV) in fruit and vegetable production water. Methods 2.5 × 10 ~ 9 pfu / mL of E.coli phage MS2100μL was added to the water sample, and the virus was captured by positive charge filter. The virus recovery efficiency was calculated according to the standard curve of E. coli phage MS2. The real-time fluorescence RT -PCR detection of hepatitis A virus in water samples, and establish a standard system of quality control. Results The HAV and E. coli bacteriophage MS2 were simultaneously added to 6 water samples for recovery of HAV. The recoveries of HAV ranged from 1.24% to 32.68% and the recoveries of MS2 ranged from 1.64% to 14.24%, reaching ISO / TS 15216-2 -2013 Virus recovery rate> 1% of the requirements. At the same time, HAV was detected in 30 real water quality samples, HAV was detected in 1 sample, the recovery rate of quality control substance Escherichia coli phage MS2 was 1.24% ~ 24.19%, and the standard deviation was 0.0612. Conclusion The established quality control system for virus detection in water samples ensures the quality of test results.