Development of subfamily-based consensus PCR assays for the detection of human and animal herpesviruses

Okoh, God'spower Richard, Lockhart, Michelle, Grimsey, Joanne, Whitmore, David, Ariel, Ellen, Butler, Jeff, and Horwood, Paul F. (2023) Development of subfamily-based consensus PCR assays for the detection of human and animal herpesviruses. European Journal of Clinical Microbiology and Infectious Diseases, 42 (6). pp. 741-746.

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Abstract

Consensus 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.

Item ID: 78700
Item Type: Article (Research - C1)
ISSN: 1435-4373
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Copyright Information: © The Author(s) 2023. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
Date Deposited: 17 May 2023 00:31
FoR Codes: 30 AGRICULTURAL, VETERINARY AND FOOD SCIENCES > 3009 Veterinary sciences > 300904 Veterinary diagnosis and diagnostics @ 100%
SEO Codes: 10 ANIMAL PRODUCTION AND ANIMAL PRIMARY PRODUCTS > 1099 Other animal production and animal primary products > 109999 Other animal production and animal primary products not elsewhere classified @ 100%
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