Author:
Okoh God’spower Richard,Lockhart Michelle,Grimsey Joanne,Whitmore David,Ariel Ellen,Butler Jeff,Horwood Paul F.
Abstract
AbstractConsensus PCR assays that can be used to sensitively detect several herpesvirus (HV) species across the different subfamilies were developed in this study. Primers containing degenerate bases were designed to amplify regions of the DNA polymerase (DPOL) gene of alpha- and gamma-HVs, and the glycoprotein B (gB) gene of beta-HVs in a singleplex, non-nested touchdown PCR format. The singleplex touchdown consensus PCR (STC-PCR) was used to amplify the DNA of eight human and 24 animal HVs. The assay was able to detect the lowest DNA dilution of 10−5 for alpha-HVs and 10−3 for beta- and gamma-HVs. In comparison, lowest detection limits of 10−5, 10−3, and 10−2 were obtained for alpha-, beta-, and gamma-HVs respectively when a nested PCR was used. The findings in this study suggest that the STC-PCR assays can be employed for the molecular surveys and clinical detection of novel and known HVs.
Funder
Australian Research Centre in Sex, Health and Society, La Trobe University
JCU Higher Degree Research Enhancement Scheme Grant
James Cook University
Publisher
Springer Science and Business Media LLC
Subject
Infectious Diseases,Microbiology (medical),General Medicine
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