摘要
目的海产品中携带病毒的核酸检测的两种前处理方法的比较。方法以市售的甲肝疫苗作为标准品,梯度稀释成不同浓度的样品,对海产品染毒后,分别采取参照GB/T22287-2008贝类中甲型肝炎病毒检测方法的前处理和使用PBS拍碎海产品消化腺这种样品前处理方法进行标本处理并提取病毒核酸,并采样荧光PCR方法检测,对甲肝病毒核酸进行定性和半定量检测,从而比较两种处理方法的差异性。结果用两种前处理方法均能检测到甲肝病毒。疫苗液、方法 1和方法 2处理的样品,其检出最高稀释倍数的lgCCID50(细胞培养半数感染量)分别为1、2和4。结论海产品的病毒核酸检测可以采用处理方法 1,该方法既简便易操作,灵敏性也优于方法 2。
Objective To compare PEG precipitation method and mashing method without PEG in viral PCR detection in sea- foods. Methods Commercial hepatitis A vaccine as standard substance were diluted to various concentrations with gradient method. Then seafoods were contaminated with the hepatitis A vaccine, and the viral nucleic acid was extracted following treat- ment by pretreatment method for hepatitis A vaccine detection of GB/T22287 -2008 and PBS mashing digestive glands, then fluorescent PCR was employed for qualitative analysis and semi - quantitative detection of hepatitis A virus severally, finally the difference of the two different pretreatment methods were compared. Results Hepatitis A virus could be detected by both meth- ods. The lgCCID50 of the highest dilution ratio were 1, 2 and 4 for vaccine solution, sample pretreated by PEG precipitation method and that by mashing method without PEG. Conclusion Pretreatment by PEG precipitationt method is better for virus PCR detection in seafoods due to easy operation and better sensitivity.
出处
《中国卫生检验杂志》
北大核心
2014年第3期372-373,共2页
Chinese Journal of Health Laboratory Technology
基金
宁波市重点科技创新团队计划资助(2012B82018)
