基于RPA-CRISPR/Cas12a的异尖线虫快速可视化检测方法

被引：1

作者：

王皓璐 ^{[1
]}

赵连静 ^{[1
]}

陈秀琴 ^{[1
]}

杨亚明 ^{[2
]}

吴方伟 ^{[2
]}

刘晓雷 ^{[1
]}

王洋 ^{[1
]}

白雪 ^{[1
]}

机构：

[1] 吉林大学动物医学学院人兽共患病研究所人兽共患病研究教育部重点实验室

[2] 云南省寄生虫病防治所

来源：

中国兽医科学 | 2024年 / 54卷 / 06期

基金：

国家重点研发计划;

关键词：

异尖线虫; RPA; CRISPR/Cas12a; 可视化检测;

D O I：

10.16656/j.issn.1673-4696.2024.0088

中图分类号：

TS254.7 [水产制品的标准与检验];

学科分类号：

摘要：

为了实现对海产品中异尖线虫的快速、便捷、高效的检测以控制其流行，针对异尖线虫内转录间隔区（ITS）保守序列设计重组酶聚合酶扩增（RPA）引物和CRISPR RNA(crRNA)，并通过优化crRNA浓度等反应条件建立RPA-CRISPR/Cas12a荧光检测体系，并评估该方法的灵敏度、特异性与重复性，通过人工污染样品验证其实际检测能力。结果表明：RPA-CRISPR/Cas12a荧光检测体系可在40 min内完成反应，敏感性为1 copy/μL，该方法仅能检测出异尖线虫，不与宫脂线虫、肠舌状绦虫、带鱼鱼肉组织、小黄花鱼鱼肉组织反应。本实验建立的RPA-CRISPR/Cas12a荧光可视化检测体系快速、灵敏、特异，可应用于海产品中异尖线虫的检测。

引用

页码：735 / 741

页数：7

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