Primers are decisive for sensitivity of PCR.

摘要: A sufficient sensitivity of PCR is a prerequisite for its use in the diagnosis infectious diseases. We have used detecting gene elements Borrelia burgdorferi, mycobacteria and Bordetella pertussis. With all these microbe groups, difficulties were encountered achieving demanded with primer pairs primarily selected. An extensive testing various reaction parameters did not improve sensitivity. Subsequently, we synthesized more primers derived from slightly different positions original target sequences. When new tested possible combinations, some reached 100-fold to 1000-fold higher than primary pairs. conclude that optimizing PCR, emphasis should be put on several screening parameters. Thus far, trial-and-error approach has used, because there no means predict properties selected pair.