公开(公告)号：US06465201B1

公开(公告)日：2002-10-15

申请号：US09574124

申请日：2000-05-19

申请人： Esther Presente ,  Barbara Young ,  Susan Upperman

发明人： Esther Presente ,  Barbara Young ,  Susan Upperman

IPC分类号： C12Q166

CPC分类号： C12Q1/48 ,  C12Q1/04 ,  C12Q1/22 ,  C12Q1/66 ,  C12Q2304/60

摘要： A method for rapidly detecting and enumerating microorganisms, having a level of microbial ATP, in the presence of mammalian cell preparations, comprising the steps of: providing a mammalian cell preparation comprising mammalian cells having a level of mammalian ATP; reducing the level of mammalian ATP in said cell preparation by, selectively lysing the mammalian cells and not lysing microbial cells with detergent or osmotic shock to extract the mammalian ATP, treating the extracted mammalian ATP with one or more ATP hydrolyzing compounds; and immobilizing the microorganisms and washing away said detergent and the hydrolyzing compound by filtering the mammalian cell preparation through a micropartioned hydrophilic/hydrophobic membrane; extracting the microbial ATP using an extracting reagent; drying the membrane; applying a bioluminescent reagent onto the membrane; and detecting and enumerating the microorganisms.

摘要翻译： 一种用于在存在哺乳动物细胞制剂的情况下快速检测和列举具有微生物ATP水平的微生物的方法，包括以下步骤：提供包含哺乳动物ATP水平的哺乳动物细胞的哺乳动物细胞制剂; 通过选择性地裂解哺乳动物细胞并且不用洗涤剂或渗透压休克裂解微生物细胞以提取哺乳动物ATP，用一种或多种ATP水解化合物处理提取的哺乳动物ATP，从而降低所述细胞制剂中哺乳动物ATP的水平; 并通过微粒亲水/疏水膜过滤哺乳动物细胞制剂，固定微生物并洗去去除所述洗涤剂和水解化合物; 使用提取试剂提取微生物ATP; 干膜; 将生物发光试剂施加到膜上; 并检测并列举微生物。