Using Droplet Digital PCR to Detect Coinfection of Human Herpesviruses 6A and 6B (HHV-6A and HHV-6B) in Clinical Samples

Ashley Vellucci; Emily C Leibovitch; Steven Jacobson

doi:10.1007/978-1-4939-7778-9_6

Using Droplet Digital PCR to Detect Coinfection of Human Herpesviruses 6A and 6B (HHV-6A and HHV-6B) in Clinical Samples

Methods Mol Biol. 2018:1768:99-109. doi: 10.1007/978-1-4939-7778-9_6.

Authors

Ashley Vellucci¹, Emily C Leibovitch^{1

2}, Steven Jacobson³

Affiliations

¹ Neuroimmunology Branch, Viral Immunology Section, National Institute of Neurological Disorders and Stroke (NINDS), National Institutes of Health, Bethesda, MD, USA.
² Institute for Biomedical Sciences, School of Medicine and Health Sciences, Washington, DC, USA.
³ Neuroimmunology Branch, Viral Immunology Section, National Institute of Neurological Disorders and Stroke (NINDS), National Institutes of Health, Bethesda, MD, USA. jacobsons@ninds.nih.gov.

PMID: 29717439
DOI: 10.1007/978-1-4939-7778-9_6

Abstract

Droplet digital™ polymerase chain reaction (ddPCR™) is a unique digital PCR technique that allows for absolute quantification of nucleic acid samples. This technique operates on the basis of amplification within water-oil emulsion droplets and can detect very small quantities of target molecules, yielding extremely precise data. Here, we describe in detail a ddPCR procedure for multiplexed detection of two clinically relevant herpesviruses, HHV-6A and HHV-6B.

Keywords: Absolute quantification; Droplet digital; HHV-6A; HHV-6B; Human herpesvirus 6; Polymerase chain reaction; Viral detection; ddPCR.

MeSH terms

Coinfection / diagnosis*
Coinfection / virology
DNA, Viral / isolation & purification*
Herpesvirus 6, Human / genetics
Herpesvirus 6, Human / isolation & purification*
Humans
Multiplex Polymerase Chain Reaction / instrumentation
Multiplex Polymerase Chain Reaction / methods*
Roseolovirus Infections / diagnosis*
Roseolovirus Infections / virology

Substances

DNA, Viral