1. Field
The present disclosure relates to methods and compositions for enhancing reaction efficiency and sensitivity in polymerase chain reaction (PCR).
2. Description of the Related Art
The number of diseases known to be caused by pathogenic microorganisms or genetic variations is increasing. For the case of rapidly progressing diseases, a precise and speedy diagnosis is desirable. For the case of diseases caused by microorganisms, the ability to detect even a trace amount is desirable because microorganism proliferation rates are high. Various methods using polymerase chain reaction (PCR) have been developed and used for early diagnosis or detection of such diseases. In particular, PCR is useful because results are obtained from a trace amount of a sample within a short time period. PCR methodology has been developed in various ways to improve detection efficiency and sensitivity. However, higher accuracy and sensitivity of PCR requires longer reaction time and higher cost. Each cycle in PCR consists of denaturation, annealing, and extension steps. In order to enhance the accuracy and sensitivity of PCR, each step in each cycle must be performed for sufficient time; and a modified polymerase is used to minimize non-specific reactions caused by the polymerase, leading to increased costs, or alternatively, a pre-activation time is added, increasing total reaction time. In addition, use of a sophisticated, high-performance apparatus leads to high costs in PCR. Depending on the sequences or structures of the primers used in PCR, annealing may occur between the primers, and as amplification proceeds, primer dimers may be generated. The primer dimers are one of the major reasons for reduction in the specificity and efficiency of PCR. Since generation of primer dimers competes with amplification of the target sequence, amplification efficiency may be significantly reduced. Generation of primer dimers may be prevented by enhancing stringency via control of the amount of salts, such as MgCl2, in the PCR reaction solution, or by using a “hot start” polymerase, e.g., hot start Taq. However, by doing so, PCR efficiency may be reduced or the total reaction time may be prolonged.
Accordingly, there is still a need to develop a PCR method capable of quickly and accurately detecting a trace amount of a target material in a sample.