Going for three - what is the role for triplex testing for HSV-1, HSV-2, and VZV

Sep 29, 2026

Uploaded by Aran

Herpes simplex virus-1 (HSV-1), HSV-2, and varicella zoster virus (VZV) commonly cause mucocutaneous lesions. While diagnostic practice has moved toward nucleic acid amplification tests (NAATs), testing is often absent or limited to virus-specific approaches based on classic clinical appearance. A recent study highlights that provider-ordered testing can miss the causative virus, affecting management and antiviral choices and differing infection-control needs across HSV-1/2 and VZV.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages4

Reading Time10 min

AccessFree to download

Abstract

Background and diagnostic shift

Limitations of virus-specific testing

Clinical impact on treatment and infection control

Need for broader triplex implementation

| Commentary Going for three: what is the role for triplex testing for HSV-1, HSV-2, and VZV? David R. Peaper1 AUTHOR AFFILIATION See affiliation list on p. 4. ABSTRACT Herpes simplex virus-1 (HSV-1), HSV-2, and varicella zoster virus (VZV) are common causes of mucocutaneous lesions. Diagnostic methods have shifted from culture or direct fluorescent antibody testing to nucleic acid amplification tests (NAATs), but testing is (i) not often undertaken or (ii) often specific for either HSV-1/2 or VZV based upon classic clinical findings associated with typical disease. A recent study by B. W. Buchan and P. Pancholi published in ASM Case Reports (2:e00112-25, 2025, [https://](https://)[ ](https://)[doi.org/10.1128/asmcr.00112-25](doi.org/10.1128/asmcr.00112-25) highlights the limitations of this approach, revealing a)[) highlights the limitations of this approach, revealing a](doi.org/10.1128/asmcr.00112-25) highlights the limitations of this approach, revealing a)[ ](doi.org/10.1128/asmcr.00112-25) highlights the limitations of this approach, revealing a)[substantial number of cases where provider-ordered testing missed the causative virus.](substantial number of cases where provider-ordered testing missed the causative virus.)[ ](substantial number of cases where provider-ordered testing missed the causative virus.)Accurate viral identification is critical in these cases, as missed or misdiagnosis impacts patient management. Treatment regimens, including antiviral dosage and duration, vary among clinical presentations of HSV-1, HSV-2, and VZV. More importantly, infection control protocols differ among these pathogens. Many cases of VZV require airborne and contact isolation in healthcare settings to prevent nosocomial transmission, a precaution not necessary for most HSV-1/2 infections. In some cases, clinical diagnosis may be insufficient for differentiation among HSV-1, HSV-2, and VZV. More studies are needed to help identify use cases where triplex testing is most appropriate, but broader availability and implementation of triplex NAATs have the potential to promote better patient management and infection prevention practices. H erpes simplex virus-1 (HSV-1), HSV-2, and varicella zoster virus (VZV) are members of Alphaherpesvirinae subfamily of the Herpesviridae family of DNA viruses (1, 2) . They are important causes of disease in both immunocompetent and immunocompromised patients, predominantly causing non-life-threatening mucocutaneous infections, but severe infections of the eye, central nervous system, liver, lungs, or disseminated infections may occur and are associated with severe morbidity and mortality. They establish latency in nerve ganglia, leading to recurrent infections and/or eruption of lesions long after primary infection (3, 4) . Seropositivity for these viruses is high, suggesting that many patients are at risk of recurrence. The clinical significance of these viruses is well established, leading to the availability of diagnostic methods, vaccines (for VZV), and antiviral medications. Traditional viral cell culture can detect HSV-1, HSV-2, and VZV, but with varying sensitivity and time to detection. Rapid viral culture and direct fluorescent antibody (DFA) testing accelerate the time to diagnosis for these pathogens, but these methods are virus specific (5, 6) . Culture and DFA have been supplanted by nucleic acid amplification tests (NAAT) due to their sensitivity and turn-around time, but virus specific testing remains the norm (5–7) . There are over 10 Food and Drug Administration (FDA)-authorized duplex tests for HSV-1 and HSV-2, a single FDA-authorized singleplex test for VZV, and only four FDA-authorized assays for triplex testing for HSV-1, HSV-2, and VZV from mucocutaneous lesions (8). Triplexing may not be compatible with some NAAT systems; Editor Nicholas M. Moore, Rush University Medical Center, Chicago, Illinois, USA Address correspondence to David R. Peaper, [david.peaper@y

FAQs

Why is identifying HSV-1, HSV-2, and VZV accurately important?

Accurate identification prevents missed or incorrect diagnoses that can alter patient management. It also determines appropriate antiviral dosage and duration and guides infection-control decisions.

How have diagnostic methods for these viruses changed?

Methods have shifted from culture and direct fluorescent antibody testing to nucleic acid amplification tests (NAATs). However, testing often remains virus-specific for HSV-1/2 or VZV rather than using triplex approaches.

What limitations does triplex testing aim to address?

Provider-ordered testing can miss the causative virus when suspicion focuses on a single pathogen. Triplex testing can support better discrimination among HSV-1, HSV-2, and VZV when clinical diagnosis is insufficient.

Sep 29, 2026

Uploaded by Aran