Differentiating microbial forensic qPCR target and control products by electrospray ionization mass spectrometry.

ABSTRACT: Molecular bioforensic research is dependent on rapid and sensitive methods such as real-time PCR (qPCR) for the identification of microorganisms. The use of synthetic positive control templates containing small modifications outside the primer and probe regions is essential to ensure all aspects of the assay are functioning properly, including the primers and probes. However, a typical qPCR or reverse transcriptase qPCR (qRT-PCR) assay is limited in differentiating products generated from positive controls and biological samples because the fluorescent probe signals generated from each type of amplicon are indistinguishable. Additional methods used to differentiate amplicons, including melt curves, secondary probes, and amplicon sequencing, require significant time to implement and validat

SUBMITTER: Motley ST 

PROVIDER: S-EPMC3696942 | biostudies-literature | 2013 Jun

REPOSITORIES: biostudies-literature