Patent ID: 6312930
Filing Date: 2001-11-06
Classification: C12Q

Abstract:
A method for the detection of a target bacteria in a complex sample mixture suspected of containing a target bacteria comprising:(i) obtaining total target bacteria DNA from said target bacteria;(ii) contacting said total target bacteria DNA with a test replication composition to form a first reaction mixture and a positive control replication composition to form a second reaction mixture, said test replication composition comprising:a) a polymeraseb) a primer pair consisting of a first primer and a second primer, each primer capable of hybridizing to a portion of said total target bacteria DNA;c) reagents and buffers necessary to effect DNA amplification;said postive control replication composition comprising:a) a polymeraseb) at least one control nucleic acid fragmentc) a single primer capable of hybridizing to a portion of said control nucleic acid fragmentd) reagents and buffers necessary to effect DNA amplification;(iii) thermocycling the first and second reaction mixtures of step (ii) thereby producing DNA amplification products consisting of either or both:a) amplified total target bacteria DNA to produce multiple copies of target DNA orb) amplified control nucleic acid fragment; and(iv) detecting the amplification products of step (iii) wherein the presence of amplified control nucleic acid fragment alone indicates a successful reaction and wherein the presence of multiple copies of target DNA indicates the presence of the target bacteria in the complex mixture.