Rapid Nucleic Acid Detection of <i>Listeria monocytogenes</i> Based on RAA-CRISPR Cas12a System
<i>Listeria monocytogenes</i> (<i>L. monocytogenes</i>) is a food-borne pathogenic bacteria that frequently contaminates animal-derived food and low-temperature preserved food. <i>Listeriosis</i> caused by its infection has a high mortality rate and poses a seriou...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-03-01
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| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/25/6/3477 |
| Summary: | <i>Listeria monocytogenes</i> (<i>L. monocytogenes</i>) is a food-borne pathogenic bacteria that frequently contaminates animal-derived food and low-temperature preserved food. <i>Listeriosis</i> caused by its infection has a high mortality rate and poses a serious threat to human health. Therefore, it is crucial to establish a sensitive, rapid and easy-to-operate technique. In this study, a Recombinase Aided Amplification (RAA) assisted CRISPR/Cas12a (RAA-CRISPR/Cas12a) fluorescence platform was established for highly sensitive nucleic acid detection of <i>L. monocytogenes</i>. The established RAA-CRISPR/Cas12a showed high sensitivity and high specificity, with the sensitivity of 350 CFU/mL and 5.4 × 10<sup>−3</sup> ng/μL for pure bacterial solution and genomic DNA, and good specificity for 5 strains of <i>Listeria</i> spp. and 14 strains of other common pathogenic bacteria. <i>L. monocytogenes</i> could be detected at an initial concentration of 2.3 CFU/25g within 2 h of enriching the beef in the food matrix, and this method could be applied to food samples that were easily contaminated with <i>L. monocytogenes</i> The results of RAA-CRISPR/Cas12a could be observed in 5 min, while the amplification was completed in 20–30 min. The speed and sensitivity of RAA-CRISPR/Cas12a were significantly higher than that of the national standard method. In conclusion, the RAA-CRISPR/Cas12a system established in this study has new application potential in the diagnosis of food-borne pathogens. |
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| ISSN: | 1661-6596 1422-0067 |