<br>基于CRISPR/Cas13a系统的BmNPV快速视觉检测技术的开发,Journal of Invertebrate Pathology

当前位置： X-MOL 学术 › J. Invertebr. Pathol. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

基于CRISPR/Cas13a系统的BmNPV快速视觉检测技术的开发
Journal of Invertebrate Pathology ( IF 3.6 ) Pub Date : 2024-02-08 , DOI: 10.1016/j.jip.2024.108072
Xue-Min Zhou ₁ , Zhen-Yu Shen ₁ , Yi-Xiang Wu ₁ , Su Lin ₁ , Meng-Dong Wang ₂ , Tao Xu ₁ , Lu-Lai Wang ₁ , Samreen Sadiq ₁ , Xin-Hao Jiao ₁ , Ping Wu ₂

Affiliation

蚕病原微生物是威胁蚕业高质量发展的重要因素。其中，家蚕核多角体病毒（BmNPV）引起的病害常导致疫情频繁、死亡率高，给蚕业造成巨大损失。目前BmNPV的分子检测方法需要昂贵的设备和熟练的技术人员。因此，蚕卵生产企业最常用的检测方法是在显微镜下观察是否存在多面体。但该方法准确度和灵敏度较低。蚕场、蚕卵生产企业和检疫部门迫切需要开发一种高灵敏度、高特异性、适用性的检测新技术。本研究将重组酶聚合酶扩增（RPA）技术与CRISPR/Cas13a系统相结合，成功建立了CRISPR/Cas13a BmNPV可视化检测技术。该技术基于微孔板横向、流动测试条和便携式荧光检测器。 30-45 min内，阳性标准质粒的检测灵敏度可达1拷贝/μL，BmNPV基因组的检测灵敏度可达1 fg/μL，具有较高的灵敏度。通过检测感染不同病原体的家蚕组织，我们确定CRISPR/Cas13a检测技术具有良好的特异性。综上所述，新建立的BmNPV核酸检测技术具有灵敏度高、特异性强、成本低、便于可视化等特点。可应用于现场检测和蚕卵质量监测系统。

"点击查看英文标题和摘要"

Development of a rapid visual detection technology for BmNPV based on CRISPR/Cas13a system

Pathogenic microorganism of silkworm are important factors that threaten the high-quality development of sericulture. Among them, Bombyx mori nucleopolyhedrovirus (BmNPV) caused diseases often lead to frequent outbreaks and high mortality, resulting in huge losses to sericultural industry. Current molecular detection methods for BmNPV require expensive equipment and sikilled technical personnel. As a result, the most commonly detection method for silkworm egg production enterprises involves observing the presence of polyhedra under a microscope. However, this method has low accuracy and sensitivity. There is an urgent need to develop a new detection technology with high sensitivity, high specificity, and applicability for silkworm farms, silkworm egg production enterprises and quarantine departments. In this study, we successfully established the CRISPR/Cas13a BmNPV visualized detection technology by combining Recombinase Polymerase Amplification (RPA) technology and CRISPR/Cas13a system. This technology is based on microplate lateral, flow test strips and portable fluorescence detector. The detection sensitivity can reach up to 1 copies/μL for positive standard plasmid and 1 fg/μL for BmNPV genome in 30–45 min, demonstrating high sensitivity. By detecting silkworm tissues infected with different pathogens, we determined that CRISPR/Cas13a detection technology has good specificity. In summary, the newly established nucleic acid detection technology for BmNPV is characterized by high sensitivity, high specificity, low cost and convenience for visualization. It can be applied in field detection and silkworm egg quality monitory system.

更新日期：2024-02-08

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南