One-step duplex RT-droplet digital PCR assay for the detection of norovirus GI and GII in lettuce and strawberry

诺如病毒 数字聚合酶链反应 生物 实时聚合酶链反应 复式(建筑) 检出限 重复性 逆转录聚合酶链式反应 分子生物学 聚合酶链反应 食品科学 病毒学 化学 色谱法 DNA 病毒 基因 遗传学 信使核糖核酸
作者
Chongzhen Sun,Jiayin Chen,Hui Li,Ling Fang,Shiwei Wu,Pallavi Jayavanth,Shuze Tang,Glòria Sánchez,Xiyang Wu
出处
期刊:Food Microbiology [Elsevier BV]
卷期号:94: 103653-103653 被引量:12
标识
DOI:10.1016/j.fm.2020.103653
摘要

The study was designed to develop a sensitive one-step duplex reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) to detect norovirus genogroup I and II (NoV GI and GII) in lettuce and strawberry. The specificity, sensitivity, repeatability and robustness of the assay was compared with RT-qPCR. The lowest concentration detected by RT-ddPCR for NoV GI and NoV GII were 4.68 and 8.47 copies/μL respectively, much lower than that of RT-qPCR with a number of 46.8 and 84.7 copies/μL, respectively. Lettuce and strawberry samples were artificially contaminated with NoV GI and GII suspensions, with inoculum size of 3.00 × 106 to 1.70 × 108 copies and 4.80 × 105 to 2.50 × 107 copies, respectively. Strawberry spiked with low inoculum size revealed positive results by RT-ddPCR, while recorded negative by RT-qPCR. Meanwhile, RT-ddPCR also showed a higher average recovery rate for NoV in lettuce and strawberry than RT-qPCR.The limit of detection (LoDs) of RT-ddPCR for NoVs in lettuce was 2.32 × 104 copies/25g (NoV GI) and 2.36 × 104 ciopies/25g (NoV GII), and that in strawberry was 2.56 × 104 copies/25g (NoV GI) and 2.64 × 104 ciopies/25g (NoV GII), which were 10 folds lower than that of RT-qPCR. The developed duplex RT-ddPCR assay exhibited stability and increased capacity to resist inhibitors in food samples with low concentration of NoV, making it a reliable method to avoid false negative result as opposed to RT-qPCR. In conclusion, one-step RT-ddPCR method developed in this study is pertinent in detecting foodborne virus such as NoV.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
1秒前
烟花应助沉海采纳,获得10
1秒前
1秒前
陈小军完成签到 ,获得积分10
2秒前
stilem完成签到,获得积分10
2秒前
libaojunok发布了新的文献求助10
2秒前
2秒前
3秒前
5秒前
张可爱完成签到,获得积分10
6秒前
orixero应助额我认为采纳,获得10
6秒前
7秒前
彭于晏发布了新的文献求助10
8秒前
8秒前
SICHEN发布了新的文献求助20
8秒前
张可爱发布了新的文献求助20
8秒前
爆米花应助眼睛大寻冬采纳,获得10
8秒前
jojo发布了新的文献求助10
10秒前
jaytotti完成签到,获得积分10
10秒前
12秒前
kanwenxian发布了新的文献求助10
13秒前
77发布了新的文献求助10
14秒前
桐桐应助顺心火龙果采纳,获得10
14秒前
小彤完成签到 ,获得积分10
15秒前
TCB发布了新的文献求助10
16秒前
leo_twli发布了新的文献求助10
16秒前
17秒前
17秒前
冷傲迎梦完成签到,获得积分20
18秒前
传奇3应助lyyyy采纳,获得10
19秒前
wu关注了科研通微信公众号
19秒前
量子星尘发布了新的文献求助10
20秒前
bkagyin应助LKF采纳,获得10
20秒前
苇一发布了新的文献求助10
21秒前
TCB完成签到,获得积分10
21秒前
21秒前
啵清啵发布了新的文献求助20
22秒前
Irony发布了新的文献求助10
22秒前
23秒前
大模型应助7275XXX采纳,获得10
24秒前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
Cognitive Neuroscience: The Biology of the Mind 1000
Technical Brochure TB 814: LPIT applications in HV gas insulated switchgear 1000
Toward a Combinatorial Approach for the Prediction of IgG Half-Life and Clearance 500
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
Picture Books with Same-sex Parented Families: Unintentional Censorship 500
Nucleophilic substitution in azasydnone-modified dinitroanisoles 500
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3970008
求助须知:如何正确求助?哪些是违规求助? 3514711
关于积分的说明 11175563
捐赠科研通 3250077
什么是DOI,文献DOI怎么找? 1795198
邀请新用户注册赠送积分活动 875630
科研通“疑难数据库(出版商)”最低求助积分说明 804931