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    METHOD FOR DETECTION AND MULTIPLE, SIMULTANEOUS QUANTIFICATION OF PATHOGENS BY MEANS OF REAL-TIME POLYMERASE CHAIN REACTION
    1.
    发明申请
    METHOD FOR DETECTION AND MULTIPLE, SIMULTANEOUS QUANTIFICATION OF PATHOGENS BY MEANS OF REAL-TIME POLYMERASE CHAIN REACTION 有权
    通过实时聚合酶链反应检测和多次同时定量病原体的方法

    公开(公告)号：US20100159448A1

    公开(公告)日：2010-06-24

    申请号：US12298298

    申请日：2007-04-19

    申请人： Elvira Garza Gonzalez ,  Francisco Javier Bosques Padilla ,  Victor Manuel Moreno Campana

    发明人： Elvira Garza Gonzalez ,  Francisco Javier Bosques Padilla ,  Victor Manuel Moreno Campana

    IPC分类号： C12Q1/68 ,  C07H21/04

    CPC分类号： C12Q1/689 ,  C12Q2600/16 ,  Y02A50/451

    摘要： A method for the detection and multiple, simultaneous quantification of any combination of Listeria sp., Staphylococcus aureus, Campylobacter jejuni and/or Escherichia coli O157.-H7, in one or more test samples, by multiplex amplification reaction, using real-time polymerase chain reaction (PCR). The steps in the method are: (a) extracting DNA from the test sample or samples; (b) preparing a reaction mixture specific for the pathogens to be detected and quantified, such that the reaction mixture contains the necessary reagents for enzymatic amplification of the extracted DNA and identification of the pathogens to be detected and quantified; (c) amplifying the reaction mixture by means of multiplex amplification reaction using PCR; and (d) simultaneously determining the presence or absence of and quantification of the pathogens in the test sample or samples. The method has the special feature that (i) the reaction mixture for enzymatic amplification of the DNA has sets of pairs of oligonucleotide primers identified as SEQ ID No. 1 and SEQ ID No. 2, SEQ ID No. 4 and SEQ ID No. 5, SEQ ID No. 7 and SEQ ID No. 8, and SEQ ID No. 10 and SEQ ID No. 11, and probes with oligonucleotide sequences identified as SEQ ID No. 3, SEQ ID No. 6, SEQ ID No. 9 and SEQ ID No. 12; (ii) the presence or absence of and quantification of said pathogens in any combination is determined by means of a fluorescent signal or fluorescence emission specific for each pathogen.

    摘要翻译： 一种用于通过多重扩增反应在一个或多个测试样品中检测和多重同时定量任何组合的利斯特氏菌属，金黄色葡萄球菌，空肠弯曲杆菌和/或大肠杆菌O157.-H7的方法，使用实时聚合酶 连锁反应（PCR）。 该方法的步骤是：（a）从测试样品或样品中提取DNA; （b）制备对待检测和定量的病原体特异性的反应混合物，使得反应混合物含有用于酶扩增提取的DNA的必需试剂和待检测和定量的病原体的鉴定; （c）通过使用PCR的多重扩增反应扩增反应混合物; 和（d）同时确定测试样品或样品中病原体的存在或不存在和定量。 该方法具有以下特征：（i）用于DNA扩增的反应混合物具有鉴定为SEQ ID No.1和SEQ ID No.2，SEQ ID No.4和SEQ ID No.的寡核苷酸引物对。 5，SEQ ID No.7和SEQ ID No.8，以及SEQ ID No.10和SEQ ID No.11的寡核苷酸序列的探针，以及鉴定为SEQ ID No.3，SEQ ID No.6，SEQ ID No. 9和SEQ ID No.12; （ii）任何组合中所述病原体的存在或不存在和定量是通过对每种病原体特异性的荧光信号或荧光发射来确定的。

    Method for detection and multiple, simultaneous quantification of pathogens by means of real-time polymerase chain reaction
    3.
    发明授权
    Method for detection and multiple, simultaneous quantification of pathogens by means of real-time polymerase chain reaction 有权
    通过实时聚合酶链反应检测和多重同时定量病原体的方法

    公开(公告)号：US08206923B2

    公开(公告)日：2012-06-26

    申请号：US12298298

    申请日：2007-04-19

    申请人： Elvira Garza Gonzalez ,  Francisco Javier Bosques Padilla ,  Victor Manuel Moreno Campana

    发明人： Elvira Garza Gonzalez ,  Francisco Javier Bosques Padilla ,  Victor Manuel Moreno Campana

    IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H21/04

    CPC分类号： C12Q1/689 ,  C12Q2600/16 ,  Y02A50/451

    摘要： A method for multiple and simultaneously detecting and quantifying of Listeria spp, Staphylococcus aureus, Campylobacter jejuni, and Escherichia coli 0157:H7, by extracting DNA from a sample; preparing a reaction mixture for enzymatic amplification of the extracted DNA and sets of pairs of oligonucleotide primers identified as SEQ ID No. 1 and SEQ ID No. 2, SEQ ID No. 4 and SEQ ID No. 5, SEQ ID No. 7 and SEQ ID No. 8, and SEQ ID No. 10 and SEQ ID No. 11, and probes with oligonucleotide sequences identified as SEQ ID No. 3, SEQ ID No. 6, SEQ ID No. 9 and SEQ ID No. 12; providing a thermostable DNA polymerase and magnesium salt to the reaction mixture; amplifying the reaction mixture by a PCR reaction; and determining the presence or absence, and quantification of the pathogens by using a fluorescent signal or fluorescence emission specific for each pathogen.

    摘要翻译： 通过从样品中提取DNA多次同时检测和定量李斯特氏菌属，金黄色葡萄球菌，空肠弯曲杆菌和大肠杆菌0157：H7的方法; 制备用于提取的DNA的酶促扩增的反应混合物和鉴定为SEQ ID No.1和SEQ ID No.2，SEQ ID No.4和SEQ ID No.5，SEQ ID No.7的寡核苷酸引物对和 SEQ ID No.8和SEQ ID No.10和SEQ ID No.11的寡核苷酸序列的探针，以及鉴定为SEQ ID No.3，SEQ ID No.6，SEQ ID No.9和SEQ ID No.12的寡核苷酸序列的探针。 向反应混合物中提供热稳定的DNA聚合酶和镁盐; 通过PCR反应扩增反应混合物; 并通过使用针对每种病原体的荧光信号或荧光发射来确定病原体的存在或不存在以及定量。