DEVELOPMENT OF A LOOP MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) ASSAY FOR IDENTIFICATION OF MEAT SPECIES

Date

2024-1-23

Author

Özden, Dilara

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Labeling meat species is crucial for sociocultural, health, and economic reasons. Therefore, developing cost-effective and precise methods is necessary. Loop-mediated isothermal amplification (LAMP) is an economical and sensitive method for visual detection. This study used LAMP for high-sensitivity identification of pork species in processed and unprocessed meats. The porcine-specific cytochrome b gene was targeted, and the LAMP assay was optimized. The Direct LAMP was used to reduce the total test time to less than an hour and eliminate the need for DNA isolation. Dual colorimetric detection was provided using hydroxynaphytol blue and cresol red dyes, improving visual detection. The cresol red detection had a sensitivity of 1 picogram, while dual colorimetric detection increased sensitivity to 0.1 picogram. Testing different kinds of processed meat samples has proven that the method successfully detects pork in processed meats. Meat mixtures with varying percentages of pork were tested, and it was found that the test sensitivity for processed and unprocessed meat was 0.1%. The developed assay was determined to be specific to pork identification by testing ten types of meats that belong to different species. The results of the developed assay were compared to the real-time PCR (polymerase chain reaction) results, and it was shown that the method could produce accurate and comparable results. It has been shown that the reaction tubes remain stable for at least six months at -20°C. Taken together, the results demonstrate that the developed LAMP assay can be used for on-site pork identification from processed and unprocessed meat samples.

Subject Keywords

LAMP, Colorimetric Detection, Porcine Detection, Direct LAMP

URI

https://hdl.handle.net/11511/108218

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Graduate School of Natural and Applied Sciences, Thesis