关键词: 国际实验动物科学协会, 能力验证, 诺如病毒

Abstract: Objective Implemented virus detection Proficiency Testing plan organized by ICLAS PEP, which contained four blind samples, and further improve laboratory testing quality. Method Analyzed and extracted nucleic acid of the samples, selected appropriate primers targeted on the promising virus to proceed polymerase chain reaction. Based on the result of polymerase chain reaction, obtain the sequencing analysis of positive result and further verify the virus in blind samples. Mixed the different dilutions of cecum contents exclude murine norovirus infection and murine norovirus culture with different concentration in the same volume, subjected to RNA extraction applied by two disparate kits and quantitative polymerase chain reaction analysis. Compared the RNA copy number derived from each experimental group with the control group. Result The four blind samples were identified as Theiler ’ s mouse encephalomyelitis virus, murine norovirus, mouse rotavirus and mouse lactate dehydrogenase hypertrophic virus separately which were agreed with the ICLAS standard result. Comparing the result of fluorescent quantitative PCR detection of norovirus in cecal contents with different dilutions, it was found that when nucleic acid was extracted with viral DNA / RNA extraction kit, the detection result showed different degrees of deviation compared with the virus control when the dilution was not higher than 1 ∶ 16, and the detection result were similar to the virus control when the dilution was 1 ∶ 32. When nucleic acid was extracted with the viral RNA extraction kit and the dilution was not less than 1 ∶ 8, the detection result was similar to the viral control. Conclusion The international comparison shows that our laboratory is reliable in virus detection, after pollution experiment research, stool samples 1 ∶ 8 diluted use viral RNA extraction kit nucleic acid for faeces, such as the best virus detection method to determine the cecum contents in samples.

Key words: ICLAS, proficiency testing, murine norovirus