摘要
目的建立鲜草莓中GⅡ型诺如病毒(No V GⅡ)的实时荧光RT-PCR检测方法,评价磁珠富集法和PEG(聚乙二醇)沉淀法对检测草莓中No V GⅡ的适用性,对北京地区采集的18份草莓样品进行检测。方法参照ISO/TS 15216-1《实时荧光RT-PCR方法测定食品中甲型肝炎病毒和诺如病毒水平方法》合成检测No V GⅡ的特异性引物和探针,分别采用磁珠富集法和PEG沉淀法富集病毒,然后提取RNA,建立实时荧光RT-PCR检测方法,并对提取条件进行优化。结果磁珠富集法的最高回收率为1.730%(PBS缓冲液),PEG沉淀法的最高回收率为1.682%(TGBE缓冲液),18份草莓样品均未检出No V GⅡ。结论通过磁珠富集法和PEG沉淀法建立的病毒检测的富集方法均适用于鲜草莓中No V GⅡ的检测。
Objective To develop a real-time PER method for detection of norovirus G Ⅱ in fresh strawberry, evaluate the application of magnetic beads enrichment method and PEG precipitation in detection of norovirus G Ⅱ and to detect norovirus G Ⅱ in 18 fresh strawberries from Beijing market. Methods Specific primers and probes were synthesized referenced to ISO/TS 15216-1. Magnetic beads and PEG precipitation were used to virus enrichment, and real-time PCR method was developed. Results The maximum recovery of magnetic beads enrichment and PEG methods were 1. 730% （ PBS buffer） and 1. 682% （TGBE buffer） respectively. All 18 samples tested were negative. Conclusion The two virus enrichment methods developed in this study were suitable for the detection of norovirus G Ⅱ in fresh strawberry.
出处
《中国食品卫生杂志》
2015年第3期242-246,共5页
Chinese Journal of Food Hygiene
基金
北京市自然科学基金重点项目"北京市售食品中诺如病毒污染
流行变异及风险评估研究"(5141002)
关键词
实时荧光RT-PCR
鲜草莓
GⅡ型诺如病毒
磁珠富集法
聚乙二醇沉淀法
Quantitative real-time PCR
fresh strawberry
uorovirus G Ⅱ
magnetic beads enrichment method
polyethylene glycol enrichment method
