Patent ID: 6013435
Filing Date: 2000-01-11
Classification: C12Q

Abstract:
A method for detecting genetic elements associated with resistance of an infectious bacteria to an antibacterial drug, the method comprising:a) providing a test sample comprising nucleic acid isolated from a clinical specimen;b) providing multiple pairs of single stranded oligonucleotide primers selected so that each pair of oligonucleotide primers is complementary to one of a plurality of target sequences of double stranded DNA associated with resistance to an antibacterial drug, said double stranded DNA of each of said target sequences having 3' ends complementary to one of the multiple pairs of oligonucleotide primers provided, and wherein one or more pairs of primers is capable of hybridizing to regions of genetic elements from different sources, and wherein said primer pairs are selected so that target sequences associated with different mechanisms of resistance to an antibacterial drug can be detected, if said target sequences are present in the test sample;c) treating the test sample with the primers under conditions such that each primer will hybridize specifically to its target sequence such that any DNA synthesis primed from nonspecific hybridization will not prevent detection of target sequences associated with resistance to the drug;d) treating the hybridized primers in a common amplification reaction mixture under conditions such that primer extension products for each pair of oligonucleotide primers are simultaneously synthesized for the sequences to which a primer is hybridized in the common amplification reaction mixture,e) repeating steps c) and d) until the target sequences present are sufficiently amplified to be detected;f) detecting the amplified target sequences, and,g) identifying the amplified target sequences correlated with the genetic elements associated with antibacterial drug resistance.