PMA-LAMP检测单增李斯特活菌方法的建立  

Establishment of PMA-LAMP for detection of viable Listeria monocytogenes

作　　者：蒋亚男[1] 满朝新[2] 赵凤[1] 刘颖[1] 薛玉清[1] 胡惠秩[1] 李博[1] 姜毓君[1,2] 

机构地区：[1]东北农业大学食品学院,黑龙江哈尔滨150030 [2]东北农业大学国家乳业工程技术研究中心,黑龙江哈尔滨150086

出　　处：《食品工业科技》2011年第7期410-412,416,共4页Science and Technology of Food Industry

基　　金：国家科技支撑计划项目(2009BADB9B06)

摘　　要：建立一种将荧光染料Propidium Monoazide(PMA)与环介导等温扩增(LAMP)相结合的检测方法,用于高效检测活的单核细胞增多性李斯特氏菌(简称单增李斯特菌)。利用PMA抑制单增李斯特死菌后进行LAMP扩增实验、并研究了PMA-LAMP方法检测单增李斯特活菌的灵敏度,同时与PMA-PCR方法灵敏度进行比较。结果表明,50μmol/L的PMA处理浓度为5×108cfu/mL单增李斯特死菌,能够完全抑制LAMP扩增。PMA-LAMP方法检测单增李斯特活菌的检出限为4.9×101cfu/mL,其灵敏度是PMA-PCR方法的10倍。该方法可以作为一种快速检测单增李斯特活菌的新技术。In order to effectively detect viable Listeria monocytogenes, a new method was developed by combination of propidium monoazide（PMA） and loop-mediated isothermal amplification（ LAMP）.The dead Listeria monocytogenes of LAMP amplification were inhibited by PMA treated, the detection sensitivity of PMA-LAMP method for viable Listeria monocytogenes was analyzed.This method was compared with the PMA-PCR method. Treated with 50μmoL PMA, LAMP amplification were inhibited.The determination limit of PMA-LAMP method was 4.9 × 10^8cfu/mL for viable Listeria monocytogenes. It＇ s sensitivity was l0 times than the PMA- PCR method. Therefore PMA-LAMP method was an effective technology with high specificity and sensitivity to detect viable Listeria monocytogenes.

关 键 词：单核细胞增多性李斯特菌 环介导等温扩增 PMA—LAMP 

分 类 号：TS207.4[轻工技术与工程—食品科学]