Dwight B. DuBois

Standards for PCR assays

Publisher Summary The wide versatility of polymerase chain reaction (PCR) has created numerous niches for this technology in research and clinical diagnostic laboratories. In both settings, well-defined DNA and RNA standards, compatible with exquisite sensitivity and specificity of PCR, are needed. Because of their relatively advanced state of development, standards for infectious agents are emphasized in this chapter. PCR-based assays have been formatted as qualitative and quantitative tools. Standards developed for quantitative assays present a special challenge as they must be precisely calibrated. Additionally, independent verification of the amount of very low numbers of nucleic acid templates is restricted by the limited number of corresponding technologies with sensitivities comparable to PCR. The chapter also outlines special problems with standardizing PCR assays. Various types of standards are also discussed including ideal, internal, and external standards. Finally, the discussion concludes by presenting the future needs for standards.