HERQ-9 Is a New Multiplex PCR for Differentiation and Quantification of All Nine Human Herpesviruses

Show full item record



Permalink

http://hdl.handle.net/10138/320479

Citation

Pyöriä , L , Jokinen , M , Toppinen , M , Salminen , H , Vuorinen , T , Hukkanen , V , Schmotz , C , Elbasani , E , Ojala , P M , Hedman , K , Välimaa , H & Perdomo , M F 2020 , ' HERQ-9 Is a New Multiplex PCR for Differentiation and Quantification of All Nine Human Herpesviruses ' , Msphere , vol. 5 , no. 3 , ARTN e00265-20 . https://doi.org/10.1128/mSphere.00265-20

Title: HERQ-9 Is a New Multiplex PCR for Differentiation and Quantification of All Nine Human Herpesviruses
Author: Pyöriä, Lari; Jokinen, Maija; Toppinen, Mari; Salminen, Henri; Vuorinen, Tytti; Hukkanen, Veijo; Schmotz, Constanze; Elbasani, Endrit; Ojala, Päivi M.; Hedman, Klaus; Välimaa, Hannamari; Perdomo, Maria F.
Contributor organization: Virus infections and immunity
Department of Virology
University of Helsinki
Klaus Hedman / Principal Investigator
Medicum
CAN-PRO - Translational Cancer Medicine Program
Faculty of Medicine
Department of Pathology
Biosciences
HUSLAB
Helsinki University Hospital Area
Department of Oral and Maxillofacial Diseases
Suu- ja leukakirurgian yksikkö
Date: 2020
Language: eng
Number of pages: 16
Belongs to series: Msphere
ISSN: 2379-5042
DOI: https://doi.org/10.1128/mSphere.00265-20
URI: http://hdl.handle.net/10138/320479
Abstract: Infections with the nine human herpesviruses (HHVs) are globally prevalent and characterized by lifelong persistence. Reactivations can potentially manifest as life-threatening conditions for which the demonstration of viral DNA is essential. In the present study, we developed HERQ-9, a pan-HHV quantitative PCR designed in triplex reactions to differentiate and quantify each of the HHV-DNAs: (i) herpes simplex viruses 1 and 2 and varicella-zoster virus; (ii) Epstein-Barr virus, human cytomegalovirus, and Kaposi's sarcoma-associated herpesvirus; and (iii) HHV-6A, -6B, and -7. The method was validated with prequantified reference standards as well as with mucocutaneous swabs and cerebrospinal fluid, plasma, and tonsillar tissue samples. Our findings highlight the value of multiplexing in the diagnosis of many unsuspected, yet clinically relevant, herpesviruses. In addition, we report here frequent HHV-DNA co-occurrences in clinical samples, including some previously unknown. HERQ-9 exhibited high specificity and sensitivity (LOD95 of similar to 10 to similar to 17 copies/reaction), with a dynamic range of 10' to 10 6 copies/p.I. Moreover, it performed accurately in the coamplification of both high- and low-abundance targets in the same reaction. In conclusion, we demonstrated that HERQ-9 is suitable for the diagnosis of a plethora of herpesvirus-related diseases. Besides its significance to clinical management, the method is valuable for the assessment of hitherto-unexplored synergistic effects of herpesvirus coinfections. Furthermore, its high sensitivity enables studies on the human virome, often dealing with minute quantities of persisting HHVs. IMPORTANCE By adulthood, almost all humans become infected by at least one herpesvirus (HHV). The maladies inflicted by these microbes extend beyond the initial infection, as they remain inside our cells for life and can reactivate, causing severe diseases. The diagnosis of active infection by these ubiquitous pathogens includes the detection of DNA with sensitive and specific assays. We developed the first quantitative PCR assay (HERQ-9) designed to identify and quantify each of the nine human herpesviruses. The simultaneous detection of HHVs in the same sample is important since they may act together to induce life-threatening conditions. Moreover, the high sensitivity of our method is of extreme value for assessment of the effects of these viruses persisting in our body and their long-term consequences on our health.
Subject: Epstein-Barr virus
HHV-6
coinfection
cytomegalovirus
diagnostics
human herpesviruses
multiplex
qPCR
quantitative methods
tonsils
viral persistence
virome
REAL-TIME PCR
EPSTEIN-BARR-VIRUS
HERPES-SIMPLEX-VIRUS
POLYMERASE-CHAIN-REACTION
VARICELLA-ZOSTER-VIRUS
INTERNATIONAL STANDARD
LIVER-TRANSPLANTATION
RAPID DETECTION
PARVOVIRUS B19
VIRAL LOAD
11832 Microbiology and virology
Peer reviewed: Yes
Rights: cc_by
Usage restriction: openAccess
Self-archived version: publishedVersion


Files in this item

Total number of downloads: Loading...

Files Size Format View
mSphere_2020_Py_ri_e00265_20.full.pdf 1.257Mb PDF View/Open

This item appears in the following Collection(s)

Show full item record