Nanoplate-based digital PCR for highly sensitive pork DNA detection targeting multi-copy nuclear and mitochondrial genes

放大器 低拷贝数 数字聚合酶链反应 生物 核DNA 线粒体DNA 基因 DNA 分子生物学 核基因 限制性酶 计算生物学 聚合酶链反应 生物化学 基因组
作者
Nur Fadhilah Khairil Mokhtar,Ying Quah Shun,Raja Kamil,Nurhidayatul Asma Mohamad,Nur Maisarah Shahidan,Irwan Hanish Warsanah,Amalia Mohd Hashim
出处
期刊:Food Additives & Contaminants: Part A [Taylor & Francis]
卷期号:41 (2): 120-133 被引量:1
标识
DOI:10.1080/19440049.2023.2298476
摘要

The inclusion of ingredients derived from pigs in highly processed consumer products poses a significant challenge for DNA-targeted analytical enforcement, which could be overcome by using digital PCR. However, most species detection methods use digital PCR to target single-copy nuclear genes, which limits their sensitivity. In this work, we examined the performance of a nanoplate-based digital PCR method that targets multi-copy nuclear (MPRE42) and mitochondrial (Cytb) genes. Poor separation of positive and negative partitions, as well as a 'rain effect' were obtained in the porcine-specific MPRE42 assay. Among the optimization strategies examined, the inclusion of restriction enzymes slightly improved the separation of positive and negative partitions, but a more extensive 'rain effect' was observed. The high copy number of the MPRE42 amplicon is hypothesized to contribute to the saturation of the positive signal. In contrast, the porcine-specific Cytb assay achieved perfect separation of positive and negative partitions with no 'rain effect'. This assay can detect as little as 0.4 pg of pork DNA, with a sensitivity of 0.05% (w/w) in a pork-chicken mixture, proving its applicability for detecting pork in meat and meat-based products. For the MPRE42 assay, potential applications in highly degraded products such as gelatin and lard are anticipated.
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
练得身形似鹤形完成签到 ,获得积分10
1秒前
1秒前
1秒前
2秒前
鹿港猫妖完成签到,获得积分20
3秒前
chen完成签到,获得积分10
3秒前
4秒前
阿湫发布了新的文献求助10
4秒前
4秒前
5秒前
崔悦欣发布了新的文献求助10
5秒前
希望天下0贩的0应助kk子采纳,获得10
6秒前
丰富的不惜完成签到,获得积分10
8秒前
票子发布了新的文献求助10
8秒前
华仔应助魏莱采纳,获得10
8秒前
F123发布了新的文献求助10
9秒前
shen发布了新的文献求助10
10秒前
10秒前
苗觉觉完成签到,获得积分10
11秒前
帕金森完成签到,获得积分10
12秒前
活力的妙芙完成签到,获得积分10
12秒前
赘婿应助疯子魔煞采纳,获得10
13秒前
橙橙橙完成签到,获得积分10
13秒前
13秒前
你好CDY完成签到,获得积分10
14秒前
14秒前
14秒前
万能图书馆应助希尔伯特采纳,获得10
14秒前
我是老大应助123采纳,获得10
15秒前
芒果小鹌鹑完成签到,获得积分10
16秒前
上官若男应助F123采纳,获得10
17秒前
Singularity应助邢文瑞采纳,获得10
18秒前
c123发布了新的文献求助10
19秒前
嘉嘉琦发布了新的文献求助10
20秒前
wanci应助QWDSA采纳,获得10
22秒前
22秒前
愉快静曼发布了新的文献求助10
25秒前
乐乐乐乐乐乐应助科学家采纳,获得10
26秒前
99完成签到,获得积分10
26秒前
26秒前
高分求助中
【提示信息,请勿应助】关于scihub 10000
Les Mantodea de Guyane: Insecta, Polyneoptera [The Mantids of French Guiana] 3000
徐淮辽南地区新元古代叠层石及生物地层 3000
The Mother of All Tableaux: Order, Equivalence, and Geometry in the Large-scale Structure of Optimality Theory 3000
Global Eyelash Assessment scale (GEA) 1000
Picture Books with Same-sex Parented Families: Unintentional Censorship 550
Research on Disturbance Rejection Control Algorithm for Aerial Operation Robots 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 4038446
求助须知:如何正确求助?哪些是违规求助? 3576149
关于积分的说明 11374627
捐赠科研通 3305875
什么是DOI,文献DOI怎么找? 1819354
邀请新用户注册赠送积分活动 892680
科研通“疑难数据库(出版商)”最低求助积分说明 815048