A rapid and sensitive Loop-mediated isothermal amplification assay for detection of pork DNA based on porcine tRNA lys and ATPase 8 genes
2017
Aliyu, S. | Igwenagu, E. | Mu’azu, A. | Rochman Naim | Abdullahi, U. F. | Wan Rohani Wan Taib
This study describes the development of a rapid and sensitive Loop-mediated isothermal amplification assay for detection of swine DNA in adulterated meat and meat products. The need to protect consumer’s right to eat foods of their choices, has made it imperative for researchers to develop efficient means of screening and certification of food products. Six sets of LAMP primers designed based on porcine tRNA lysine gene and ATPase subunit 8 genes were used for the assay. Amplification was carried out under constant temperature (630C), using a simple laboratory water bath. Average time spent in amplification and detection of results was25 min. All results were visually detected and confirmed by electrophoresis. Detection limit of the assay was 0.03 femtogram (fg) much high than the PCR assay, and detection probability ofthe assay was 100%. Detection of 0.5% of pork spiked with 99.5% of cattle beef is indicative of the sensitivity and robustness of the assay. This could serve as a prototype for development of a sensitive and inexpensive Swine DNA LAMP detection kit.
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