Wang Jieru, Wang Nan, Xu Lei, Zeng Xiaoyu, Cheng Junsheng, Zhang Xiaoqian, Zhang Yinghui, Yin Dongdong, Gou Jiaojiao, Pan Xiaocheng, Zhu Xiaojie
Anhui Province Key Laboratory of Livestock and Poultry Product Safety Engineering, Livestock and Poultry Epidemic Diseases Research Center of Anhui Province, Key Laboratory of Pig Molecular Quantitative Genetics of Anhui Academy of Agricultural Sciences, Institute of Animal Husbandry and Veterinary Sciences, Anhui Academy of Agricultural Sciences, Hefei 230031, China.
China Institute of Veterinary Drug Control, Beijing 100000, China.
Foods. 2024 May 21;13(11):1601. doi: 10.3390/foods13111601.
(), the microorganism responsible for bovine tuberculosis (bTB), is transferred to people by the ingestion of unpasteurized milk and unprocessed fermented milk products obtained from animals with the infection. The identification of in milk samples is of the utmost importance to successfully prevent zoonotic diseases and maintain food safety. This study presents a comprehensive description of a highly efficient molecular test utilizing recombinase-aided amplification (RPA)-clustered regularly interspaced short palindromic repeat (CRISPR)-associated protein (Cas) 13a-lateral flow detection (LFD) for detection. In contrast to ELISA, RPA-CRISPR-Cas13a-LFD exhibited greater accuracy and sensitivity in the detection of in milk, presenting a detection limit of 2 × 10 copies/μL within a 2 h time frame. The two tests exhibited a moderate level of agreement, as shown by a kappa value of 0.452 (95%CI: 0.287-0.617, < 0.001). RPA-CRISPR-Cas13a-LFD holds significant potential as a robust platform for pathogen detection in complex samples, thereby enabling the more dependable regulation of food safety examination, epidemiology research, and medical diagnosis.
牛型结核分枝杆菌（Mycobacterium bovis）是牛结核病（bTB）的病原体，通过摄入来自受感染动物的未经巴氏杀菌的牛奶和未加工的发酵奶制品传染给人类。鉴定牛奶样本中的牛型结核分枝杆菌对于成功预防人畜共患病和维持食品安全至关重要。本研究全面描述了一种高效的分子检测方法，该方法利用重组酶辅助扩增（RPA）-成簇规律间隔短回文重复序列（CRISPR）-相关蛋白（Cas）13a-侧向流动检测（LFD）来检测牛型结核分枝杆菌。与酶联免疫吸附测定（ELISA）相比，RPA-CRISPR-Cas13a-LFD在检测牛奶中的牛型结核分枝杆菌时表现出更高的准确性和灵敏度，在2小时内检测限为2×10拷贝/微升。如kappa值为0.452（95%CI：0.287-0.617，P<0.001）所示，这两种检测方法的一致性处于中等水平。RPA-CRISPR-Cas13a-LFD作为一种强大的复杂样本病原体检测平台具有巨大潜力，从而能够更可靠地监管食品安全检查、流行病学研究和医学诊断。
