TY - JOUR AU - Tran, Hau Thi AU - Tran, Diem Hong AU - Pham, Trang Nguyen Minh AU - Phung, Huong Thi Thu PY - 2022/04/01 Y2 - 2024/03/29 TI - DIRECT RECOMBINASE POLYMERASE AMPLIFICATION ASSAY FOR ACCURATE AND RAPID DETECTION OF LISTERIA MONOCYTOGENES IN FOOD JF - Journal of microbiology, biotechnology and food sciences JA - J microb biotech food sci VL - 11 IS - 5 SE - Food Sciences DO - 10.55251/jmbfs.4749 UR - https://office2.jmbfs.org/index.php/JMBFS/article/view/4749 SP - e4749 AB - <p><em>Listeria monocytogenes </em>is one of the most common types of food poisoning bacteria which can cause serious foodborne diseases or even lethality. Generally, <em>L. monocytogenes </em>can be detected using traditional microbiology or molecular biology techniques, notably PCR. However, the application of these methods at the field is restricted due to the strict requirement of equipment and skilled personnel. In this study, recombinase polymerase amplification (RPA), an isothermal PCR assay was developed to rapidly detect <em>L. monocytogenes </em>in 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 <em>L. monocytogenes </em>genomic DNA, which was 1000-fold more sensitive than the conventional PCR. RPA also succeeded to directly detect <em>L. monocytogenes </em>cells at a concentration as low as 2.5 × 10<sup>1</sup> Colony Forming Unit (CFU)/mL in pure cultures. In addition, RPA could accurately detect <em>L. monocytogenes </em>at 2.5 × 10<sup>2</sup> 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 <em>L. monocytogenes </em>in food. Accordingly, this rapid and sensitive method could be further applied to clinical testing for the diagnosis of <em>L. monocytogenes </em>infection, especially in areas with limited settings.</p> ER -