Integrating loop-mediated isothermal amplification with lateral flow assay to achieve a highly sensitive method for detecting Streptococcus suis Genome in raw pork
Eakkapote Prompunt,
Weeraya Thongkum,
Thitima Sumphanapai,
Parin Kamseng,
Somphot Saoin,
Chiraphat Kloypan,
Chatchai Tayapiwatana,
Sawitree Nangola
Affiliations
Eakkapote Prompunt
Division of Clinical Microbiology and Medical Parasitology, Department of Medical Technology, School of Allied Health Sciences, University of Phayao, Phayao, 56000, Thailand
Weeraya Thongkum
Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand; Center of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand; Center of Innovative Immunodiagnostic Development, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand
Thitima Sumphanapai
Division of Clinical Hematology and Microscopy, Department of Medical Technology, School of Allied Health Sciences, University of Phayao, Phayao, 56000, Thailand
Parin Kamseng
Division of Clinical Hematology and Microscopy, Department of Medical Technology, School of Allied Health Sciences, University of Phayao, Phayao, 56000, Thailand
Somphot Saoin
Division of Clinical Immunology and Transfusion Sciences, Department of Medical Technology, School of Allied Health Sciences, University of Phayao, Phayao, 56000, Thailand
Chiraphat Kloypan
School of Medicine, University of Phayao, Phayao, 56000, Thailand
Chatchai Tayapiwatana
Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand; Center of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand; Center of Innovative Immunodiagnostic Development, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand
Sawitree Nangola
Division of Clinical Immunology and Transfusion Sciences, Department of Medical Technology, School of Allied Health Sciences, University of Phayao, Phayao, 56000, Thailand; Corresponding author.
Streptococcus suis (S.suis), 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 S. suis in raw pork was developed by integrating loop-mediated isothermal amplification (LAMP) and a lateral flow assay (LFA), S. suis LAMP-LFA. LAMP reactions targeting the S. suis glutamate dehydrogenase (gdh) gene were optimized for specific detection of S. suis 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 S. suis 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 101 CFU per 25 g of raw pork. S. suis 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 S. suis detection in raw pork, contributing to enhanced food safety and public health protection.
