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专利名称：METHOD FOR DETECTION AND MULTIPLE,
SIMULTANEOUS QUANTIFICATION OFPATHOGENS BY MEANS OF REAL-TIMEPOLYMERASE CHAIN REACTION
发明人：GARZA GONZÁLEZ, Elvira,BOSQUES
PADILLA, Francisco Javier,MORENOCAMPAÑA, Victor Manuel
申请号：EP07747198.5申请日：20070419公开号：EP2020449A1公开日：20090204
摘要：A method for the detection and multiple, simultaneous quantification of anycombination of Listeria sp., Staphylococcus aureus, Campylobacter jejuni and/orEscherichia coli O157.-H7, in one or more test samples, by multiplex amplificationreaction, 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 mixturespecific for the pathogens to be detected and quantified, such that the reaction mixturecontains the necessary reagents for enzymatic amplification of the extracted DNA andidentification of the pathogens to be detected and quantified; (c) amplifying the reactionmixture by means of multiplex amplification reaction using PCR; and (d) simultaneouslydetermining the presence or absence of and quantification of the pathogens in the testsample or samples. The method has the special feature that (i) the reaction mixture forenzymatic 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 IDNo. 8, and SEQ ID No. 10 and SEQ ID No. 11, and probes with oligonucleotide sequencesidentified as SEQ ID No. 3, SEQ ID No. 6, SEQ ID No. 9 and SEQ ID No. 12; (ii) the presenceor absence of and quantification of said pathogens in any combination is determined bymeans of a fluorescent signal or fluorescence emission specific for each pathogen.
申请人：Sigma Alimentos, S.A. De C.V.
地址：Ave. Gomez Morin 1111 Co. Carrizalejo, C.P. 66254 San Pedro Garza GarciaNeuvo Leon MX
国籍：MX
代理机构：Elzaburu Marquez, Alberto
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