寵物食品中肉毒桿菌LAMP檢測方法的研究
發(fā)布時間:2018-06-08 23:52
本文選題:肉毒桿菌 + LMAP技術(shù)。 參考:《中國畜牧雜志》2017年04期
【摘要】:為研究寵物食品中肉毒桿菌環(huán)介導(dǎo)恒溫擴增(LAMP)檢測方法,本試驗根據(jù)肉毒桿菌特異性基因片段(Ntn H)設(shè)計1組引物,應(yīng)用LMAP技術(shù),通過擴增肉毒桿菌和14種非肉毒桿菌基因片段,對建立的LAMP反應(yīng)體系進行最佳溫度及反應(yīng)時間篩選試驗、采用濁度法和染色法進行特異性試驗以及與PCR對比的敏感性試驗。結(jié)果表明:建立的LAMP反應(yīng)體系只擴增肉毒桿菌基因,特異性良好;體系最佳反應(yīng)溫度為64℃,最佳反應(yīng)時間為60 min;最低檢測濃度為0.0001 pg/μL,比PCR高出10倍。本試驗所建立的方法特異性好、靈敏度高,反應(yīng)時間短,適用于寵物食品中肉毒桿菌的檢測。
[Abstract]:In order to study the detection method of botulinum loop mediated isothermal amplification (Lampp) in pet food, a group of primers was designed according to the specific gene fragment of Botox and 14 kinds of non-botulinum gene fragments were amplified by LMAP technique. The optimum temperature and reaction time of the lamp reaction system were screened, the specificity test and the sensitivity test compared with PCR were carried out by turbidimetric method and staining method. The results showed that the lamp reaction system only amplified the Botox gene with good specificity, the optimum reaction temperature was 64 鈩,
本文編號:1997796
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