Prompunt Eakkapote, Thongkum Weeraya, Sumphanapai Thitima, Kamseng Parin, Saoin Somphot, Kloypan Chirapat, Tayapiwatana Chatchai, Nangola Sawitree
Division of Clinical Microbiology and Medical Parasitology, Department of Medical Technology, School of Allied Health Sciences, University of Phayao, Phayao, 56000, Thailand.
Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand.
Heliyon. 2024 Aug 28;10(17):e36942. doi: 10.1016/j.heliyon.2024.e36942. eCollection 2024 Sep 15.
., a zoonotic foodborne pathogen prevalent in Southeast Asia, poses a substantial threat to human and animal health because of its ability to cause severe and life-threatening illnesses. To address this challenge, a rapid and highly sensitive detection platform for in raw pork was developed by integrating loop-mediated isothermal amplification (LAMP) and a lateral flow assay (LFA), LAMP-LFA. LAMP reactions targeting the glutamate dehydrogenase () gene were optimized for specific detection of within 45 min at an isothermal temperature of 65 °C. The assay exhibited marked sensitivity, with a detection limit of 100 fg for genomic DNA extracted from cultures. Notably, this method showed no cross-reactivity with other bacterial contaminants commonly found in raw pork. The resulting LAMP amplicons were effectively detected using LFA, with a test limit of 10 CFU per 25 g of raw pork. LAMP-LFA proved to be highly specific and reliable, with no false-positives detected in spiked pork samples or pork samples containing other bacterial contaminants. Due to its high sensitivity, specificity, and rapid turnaround time, the proposed technique has immense potential as a field-deployable screening test for detection in raw pork, contributing to enhanced food safety and public health protection.
……是一种在东南亚流行的人畜共患食源性病原体，因其能够引发严重且危及生命的疾病，对人类和动物健康构成了重大威胁。为应对这一挑战，通过整合环介导等温扩增技术（LAMP）和侧向流动分析（LFA），即LAMP-LFA，开发了一种用于生猪肉中……的快速且高度灵敏的检测平台。针对谷氨酸脱氢酶（……）基因的LAMP反应进行了优化，可在65℃等温温度下45分钟内特异性检测……。该检测方法具有显著的灵敏度，从……培养物中提取的基因组DNA的检测限为100 fg。值得注意的是，该方法与生猪肉中常见的其他细菌污染物无交叉反应。使用LFA可有效检测LAMP扩增产物，每25克生猪肉的检测限为10 CFU。LAMP-LFA被证明具有高度特异性和可靠性，在加标猪肉样品或含有其他细菌污染物的猪肉样品中未检测到假阳性。由于其高灵敏度、特异性和快速周转时间，所提出的技术作为一种可现场部署的生猪肉中……检测筛选试验具有巨大潜力，有助于加强食品安全和公共卫生保护。
