RESUMEN
Objective To establish a accurate and rapid method of loop-mediated isothermal amplification(LAMP) for detecting Legionella Pneumophila(LP) .Methods The strains of LP ,Staphylococcus aureus ,Pseudomonas aeruginosa ,Escherichia coli ,Enterobacter sakazakii ,Listeria monocytogenes ,Salmonella typhimurium ,shigella flexneri and Vibrio parahaemolyticus were selected . According to six special domains of toxicity-related mip gene on LP ,the LAMP primers(mip-1 ,mip-2 ,mip-3) were designed by using the Primer Explorer Version 4 .0 .The genomic DNA was extracted for conducting LAMP .Then its specificity ,lowest detectable limit and stability were evaluated .Results The screened primer mip-3 appeared the peak after amplification for about 10 min in the LAMP reaction system for detecting LP ,moreover the peak value was higher ;while the strains of non-LP had no amplification reaction ;the LAMP detection limit could reach 100 fg/μL .The primer mip-3 appeared the peak almost at the same time in 20 times of duplicated detection ,and its stability was good .Conclusion The established LAMP detection method has the advantages of strong specificity and high stability ,can rapidly and accurately detect LP and has large prospect of promotion and application .
RESUMEN
Ochratoxin A (OTA) is a toxic secondary metabolite mainly produced by Aspergillus and Penicillium species, with strong renal toxicity, teratogenic, carcinogenic, mutagenic effect. Studies have shown that OTA is not only widely contaminated in food and feed crops, but also has been widely contaminated in Chinese herbal medicines such as spices, licorice and so on. In view of OTA's universality and harmfulness, this paper summarizes the flow visualization test strip, microsphere, electrochemical sensor, surface enhanced Raman spectroscopy technology in OTA rapid detection, which provides reference for the research and application of high throughout detection instrument miniaturization in order to achieve OTA quick detection and simple operation.