tailieunhanh - Direct recombinase polymerase amplification assay for accurate and rapid detection of listeria monocytogenes in food

The results showed that the RPA reaction, without requiring complex thermal cycles, was well-performed in the optimal conditions of 39°C within only 25 minutes. The limit of detection was identified as 310 fg of L. monocytogenes genomic DNA, which was 1000-fold more sensitive than the conventional PCR. RPA also succeeded to directly detect L. monocytogenes cells at a concentration as low as × 101 Colony Forming Unit (CFU)/mL in pure cultures. In addition, RPA could accurately detect L. monocytogenes at × 102 CFU/mL in milk without sample extraction or processing. | DIRECT RECOMBINASE POLYMERASE AMPLIFICATION ASSAY FOR ACCURATE AND RAPID DETECTION OF LISTERIA MONOCYTOGENES IN FOOD Hau Thi Tran Diem Hong Tran Trang Nguyen Minh Pham Huong Thi Thu Phung Address es NTT Hi-Tech Institute Nguyen Tat Thanh University 298A Nguyen Tat Thanh Ho Chi Minh City 700000 Vietnam 84981411701. Corresponding author ptthuong@ https ARTICLE INFO ABSTRACT Received 7. 5. 2021 Listeria monocytogenes is one of the most common types of food poisoning bacteria which can cause serious foodborne diseases or even lethality. Generally L. monocytogenes can be detected using traditional microbiology or molecular biology techniques notably PCR. Revised 9. 11. 2021 However the application of these methods at the field is restricted due to the strict requirement of equipment and skilled personnel. In Accepted 9. 11. 2021 this study recombinase polymerase amplification RPA an isothermal PCR assay was developed to rapidly detect L. monocytogenes in Published crude samples. The results showed that the RPA reaction without requiring complex thermal cycles was well-performed in the optimal conditions of 39 C within only 25 minutes. The limit of detection was identified as 310 fg of L. monocytogenes genomic DNA which Regular article was 1000-fold more sensitive than the conventional PCR. RPA also succeeded to directly detect L. monocytogenes cells at a concentration as low as 101 Colony Forming Unit CFU mL in pure cultures. In addition RPA could accurately detect L. monocytogenes at 102 CFU mL in milk without sample extraction or processing. Therefore RPA established in this study could be an alternative standard method to confirm the presence of L. monocytogenes in food. Accordingly this rapid and sensitive method could be further applied to clinical testing for the diagnosis of L. monocytogenes infection especially in areas with limited settings. Keywords Listeria monocytogenes direct RPA foodborne diseases .