Hu Zhun, Qin Tao, Qian Luyao, Xu Lu, Zhang Liwu, Deng Shuangsheng, Tao Jianping, Hu Junjie
Yunnan Key Laboratory for Plateau Mountain Ecology and Restoration of Degraded Environments, School of Ecology and Environmental Sciences, Yunnan University, Kunming 650091, China.
Chongqing TopMe Bio-Engineering Co., Ltd., Chongqing 422460, China.
Animals (Basel). 2025 Sep 22;15(18):2763. doi: 10.3390/ani15182763.
The domestic pig () serves as an intermediate host for two species: the non-zoonotic and the zoonotic , both of which threaten animal and human health and contribute to economic losses in swine production. Existing diagnostic methods, such as microscopy and PCR, suffer from limitations regarding their sensitivity, cost, and field applicability, especially in resource-constrained settings. To address these challenges, we developed a highly specific and ultrasensitive loop-mediated isothermal amplification (LAMP) assay targeting the mitochondrial cytochrome c oxidase subunit I (1) gene for rapid detection. The optimized protocol (62 °C for 40 min) showed absolute specificity, with no cross-reactivity to related coccidia. The assay exhibited remarkable sensitivity, detecting as little as 6.7 × 10 ng/μL () and 5.4 × 10 ng/μL (), representing 10-fold and 10,000-fold improvements over conventional PCR, respectively. With a simple visual readout, the proposed LAMP assay eliminates the need for sophisticated equipment, making it an ideal field-deployable diagnostic tool for basic laboratories and under-resourced regions.
家猪（）是两种球虫的中间宿主：非人畜共患的和人畜共患的，这两种球虫都威胁动物和人类健康，并导致猪生产中的经济损失。现有的诊断方法，如显微镜检查和聚合酶链反应（PCR），在灵敏度、成本和现场适用性方面存在局限性，尤其是在资源有限的环境中。为应对这些挑战，我们开发了一种高度特异性和超灵敏的环介导等温扩增（LAMP）检测方法，该方法靶向线粒体细胞色素c氧化酶亚基I（1）基因以进行快速检测。优化后的方案（62℃，40分钟）显示出绝对特异性，与相关球虫无交叉反应。该检测方法具有显著的灵敏度，分别能检测低至6.7×10 ng/μL（）和5.4×10 ng/μL（），比传统PCR分别提高了10倍和10000倍。通过简单的视觉读数，所提出的LAMP检测方法无需复杂设备，使其成为基础实验室和资源匮乏地区理想的可现场部署的诊断工具。
