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

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.

Herpesvirus Diseases in Humans and Animals: Recent Developments, Challenges, and Charting Future Paths

“…They cause a spectrum of diseases, ranging from mild, self-limiting conditions to severe, life-threatening illnesses. With the ability to establish lifelong relationships with their hosts and trigger recurrent outbreaks, herpesviruses have emerged as formidable players in the realm of infectious diseases [1][2][3]. Human herpesviruses, including herpes simplex virus (HSV), varicella-zoster virus (VZV), cytomegalovirus (CMV), Epstein-Barr virus (EBV), Kaposi's sarcoma-associated herpesvirus (KSHV), and others, continue to exert a substantial impact on global public health [4,5].…”