Linear After The Exponential (LATE) Polymerase Chain Reaction: Potential for Clinical Diagnostics

Abstract

The linear-after-the-exponential polymerase chain reaction (LATE-PCR) is an advanced asymmetric PCR method that generates single-stranded DNA amplicons under unique amplification conditions that involve unequal concentrations of limiting and excess primers. When the limiting primer is depleted, the excess primer will continue to form single-stranded DNA (ssDNA) products, in contrast to conventional PCR. Additionally, LATE-PCR potentially has increased allele discrimination because of the wide temperature probing window between mis-matched and matched targets. Mismatch-tolerant and temperature-dependent probe use enables highly multiplexing amplification. The LATE-PCR assay provides detection capability for single cell diagnosis of congenital diseases, genotyping, sequencing, and serotypes of viruses causing disease in animal. LATE-PCR has demonstrated higher sensitivity and precision than conventional PCR. Rapid, multiplex LATE-PCR has the potential to improve performance of nucleic acid detection and may be a useful diagnostic tool at the point of care.