Molecular Detection of Methicillin-resistant Staphylococcus aureus Isolated From Foods in Germany Using LAMP Assay

Abstract

HE existence of methicillin-resistant Staphylococcus aureus (MRSA) represents a considerable a major danger to the public health, contributing to both foodborne illnesses and hospital-acquired infections. Hundred eighty-nine of Staphylococcus aureus (S. aureus) isolates were obtained from diverse food items, including poultry (103 isolates), sausage (9 isolates), pork (3 isolates), and minced meat (4 isolates), along with bovine mastitis milk (70 isolates). These isolates were submitted to the University of Veterinary Medicine Hannover, Germany, as part of routine diagnostic procedures for S. aureus. This study utilized numerous S. aureus isolates from food sources to develop an efficient the loop-mediated isothermal amplification (LAMP) technique enables the swift identification of S. aureus and MRSA. Six primers were specifically crafted for each S. aureus and MRSA sequence. Results indicated that all presumed S. aureus isolates were successfully identified with the nuc gene through LAMP and PCR techniques. However, 8.46% of isolates were MRSA which possessed the mecA gene. The LAMP assay exhibited high analytical sensitivity (1.49 pg/µL-1, 100% sensitivity) for detecting MRSA DNA with a dilution factor of 10-4. We determined the Limit of Detection (LOD) as 3.6 x 103 CFU/mL, with an 80% detection probability. This method can be seamlessly approved into any microbiology laboratory for mecA detection, offering the advantages of simplicity, time efficiency, and accurate results without the need for specialized equipment.