PCR METHOD USING MULTIPLE PRIMER PAIRS AND THE SOLUTION USED IN IT AND THE USE OF THE METHOD IN THE DETECTION REAGENTS

摘要

The invention relates to a method of polymerase chain reaction which use multiple primer pairs and also relates to the solution is used in it. Specifically, the PCR reaction was divided into three phases. During the first phase's 2-25 cycles, annealing temperature varies from 60-80°C. During the third phase's 5-25 cycles; denaturing temperature varies from 65-87°C. The second phase, which consists of 2-4 cycles, exists between the first and the third phase. The annealing temperature of this phase is same with that of the first phase. The method of the invention has overcome the disadvantage in the art that non-specific amplification between the sense and antisence primers and the nonspecific amplification product thus obtained in the multiplex PCR. The method is suitable for the preparation of the rapid detect agents that used in single-tube multiplex PCR and it also has good application future in the detection of the virus, such as HBV, HCV, HIV, HPV and so on, and the variants, stereotype, genotype or the subtype thereof, and in the detection of drug resistant gene of the bacterium such as tubercle bacillus.