title: A novel low temperature PCR assured high-fidelity DNA amplification.
abstract: As previously reported, a novel low temperature (LoTemp) polymerase chain reaction (PCR) catalyzed by a moderately heat-resistant (MHR) DNA polymerase with a chemical-assisted denaturation temperature set at 85 °C instead of the conventional 94-96 °C can achieve high-fidelity DNA amplification of a target DNA, even after up to 120 PCR thermal cycles. Furthermore, such accurate amplification is not achievable with conventional PCR. Now, using a well-recognized L1 gene segment of the human papillomavirus (HPV) type 52 (HPV-52) as the template for experiments, we demonstrate that the LoTemp high-fidelity DNA amplification is attributed to an unusually high processivity and stability of the MHR DNA polymerase whose high fidelity in template-directed DNA synthesis is independent of non-existent 3'-5' exonuclease activity. Further studies and understanding of the characteristics of the LoTemp PCR technology may facilitate implementation of DNA sequencing-based diagnostics at the point of care in community hospital laboratories.
authors.affiliation: Milford Hospital and Milford Molecular Laboratory, 2044 Bridgeport Avenue, Milford, CT 06460, USA. gfhong@sibcb.ac.cn.
authors.forename: Sin Hang
authors.initials: SH
authors.lastname: Lee
authors.forename: Shichao
authors.initials: S
authors.lastname: Ge
authors.forename: Shaoxia
authors.initials: S
authors.lastname: Zhou
authors.forename: Guofan
authors.initials: G
authors.lastname: Hong
mesh_terms: D020137:Base Pair Mismatch / Q000235:genetics; D001483:Base Sequence; D003080:Cold Temperature; D004247:DNA / Q000378:metabolism*; D017931:DNA Primers / Q000378:metabolism; D005814:Genes, Viral; D006801:Humans; D007700:Kinetics; D027383:Papillomaviridae / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000378:metabolism
chemical_list: D017931:DNA Primers; D014764:Viral Proteins; D004247:DNA