Duplex Real-Time Polymerase Chain Reaction Assay for Detection and Quantification of Herpes Simplex Virus Type 1 and Herpes Simplex Virus Type 2 in Genital and Cutaneous Lesions

Filén, Finn; Strand, Anders; Allard, Annika; Blomberg, Jonas; Herrmann, Björn

doi:10.1097/00007435-200406000-00002

Cited by 55 publications

(42 citation statements)

References 24 publications

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“…The principle of qPCR has been described elsewhere (27), as has the design of primers and probes for HSV-1 and HSV-2 detection (20). In brief, the qPCR was carried out using a degenerate primer pair specific for the glycoprotein G gene (HSV-2) and both the G glycoprotein gene and the J glycoprotein gene (HSV-1): forward primer (HSV-1, 5=-GGC CTG GCT ATC CGG AGA-3=; HSV-2, 5=-AGA TAT CCT CTT TAT CAT CAG CAC CA-3=) and reverse primer (HSV-1, 5=-GCG CAG AGA CAT CGC GA-3=; HSV-2, 5=-TTG TGC TGC CAA GGC GA-3=).…”

Section: Figmentioning

confidence: 99%

2-[4,5-Difluoro-2-(2-Fluorobenzoylamino)-Benzoylamino]Benzoic Acid, an Antiviral Compound with Activity against Acyclovir-Resistant Isolates of Herpes Simplex Virus Types 1 and 2

Strand

Islam

Edlund

et al. 2012

Antimicrob Agents Chemother

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e Herpes simplex viruses 1 and 2 (HSV-1 and HSV-2) are responsible for lifelong latent infections in humans, with periods of viral reactivation associated with recurring ulcerations in the orofacial and genital tracts. In immunosuppressed patients and neonates, HSV infections are associated with severe morbidity and, in some cases, even mortality. Today, acyclovir is the standard therapy for the management of HSV infections. However, the need for novel antiviral agents is apparent, since HSV isolates resistant to acyclovir therapy are frequently isolated in immunosuppressed patients. In this study, we assessed the anti-HSV activity of the antiadenoviral compounds 2-[2-(2-benzoylamino)-benzoylamino]benzoic acid (benzavir-1) and 2-[4,5-difluoro-2-(2-fluorobenzoylamino)-benzoylamino]benzoic acid (benzavir-2) on HSV-1 and HSV-2. Both compounds were active against both viruses. Importantly, benzavir-2 had potency similar to that of acyclovir against both HSV types, and it was active against clinical acyclovir-resistant HSV isolates.

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Section: Figmentioning

confidence: 99%

2-[4,5-Difluoro-2-(2-Fluorobenzoylamino)-Benzoylamino]Benzoic Acid, an Antiviral Compound with Activity against Acyclovir-Resistant Isolates of Herpes Simplex Virus Types 1 and 2

Strand

Islam

Edlund

et al. 2012

Antimicrob Agents Chemother

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“…While previous reports have shown that laboratory-developed PCR assays are more sensitive than culture for the diagnosis of HSV in dermal and genital samples (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11), no prior studies have analyzed the performance characteristics of the Simplexa HSV 1 & 2 direct PCR assay (Focus Diagnostics, Cypress, CA), a new commercial assay that can be performed directly on clinical samples in a nonmolecular laboratory.…”

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confidence: 99%

Comparison of Simplexa HSV 1 & 2 PCR with Culture, Immunofluorescence, and Laboratory-Developed TaqMan PCR for Detection of Herpes Simplex Virus in Swab Specimens

2013

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“…The quantification of HSV genomes by real-time PCR in genital fluid has been previously reported in a few studies (3,16,34,35,41), but no published procedure has been validated yet. We have herein validated a laboratory-developed one-step real-time PCR assay to quantify accurately HSV DNA in cervicovaginal fluid by using as standard a plasmid containing HSV DNA polymerase target.…”

Section: Discussionmentioning

confidence: 99%

Real-Time PCR Quantification of Genital Shedding of Herpes Simplex Virus (HSV) and Human Immunodeficiency Virus (HIV) in Women Coinfected with HSV and HIV

Goff

Bouhlal

Grésenguet³

et al. 2006

J Clin Microbiol

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The accuracy and usefulness of laboratory-developed real-time PCR procedures using a Light Cycler instrument (Roche Diagnostics) for detecting and quantifying human immunodeficiency virus type 1 (HIV-1) RNA and DNA as well as herpes simplex virus type 1 (HSV-1)/HSV-2 DNA in cervicovaginal secretions from women coinfected with HIV and HSV were evaluated. For HIV-1, the use of the NEC152 and NEC131 primer set and the NEC-LTR probe in the long terminal repeat gene allowed us to detect accurately the majority of HIV-1 subtypes of group M circulating in sub-Saharan Africa, including subtypes A, B, C, D, and G as well as circulating recombinant forms 02 and 11. The detection threshold of real-time PCR for HIV in cervicovaginal lavage samples was 5 copies per assay for both RNA and DNA; the intra-and interassay coefficients of variation of C T values were 1.30% and 0.69% (HIV-1 RNA) and 1.84% and 0.67% (HIV-1 DNA), respectively. Real-time PCR for HSV using primers and probe targeting the HSV DNA polymerase gene allowed both detection and quantification of HSV DNA and also differentiation between HSV-1 and HSV-2 genotypes. The detection threshold of real-time PCR for HSV was 5 copies per assay; the intra-and interassay coefficients of variation of C T values were 0.96% and 1.49%, respectively. Both manual and automated silica-based procedures were appropriate for combined extraction of HIV and HSV genomes from female genital secretions. Taken together, these findings indicate that real-time PCR may be used as a unique nucleic acid amplification procedure to detect and quantify HIV and HSV genomes in cervicovaginal secretions and thus to assess at reduced costs the genital shedding of both viruses in women included in intervention studies.

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Duplex Real-Time Polymerase Chain Reaction Assay for Detection and Quantification of Herpes Simplex Virus Type 1 and Herpes Simplex Virus Type 2 in Genital and Cutaneous Lesions

Abstract: Real-time PCR is a sensitive method for diagnosing genital herpes, and the duplex format is convenient for typing. The method increased the detection rate by 27% compared with virus culture.

Cited by 55 publications

References 24 publications

2-[4,5-Difluoro-2-(2-Fluorobenzoylamino)-Benzoylamino]Benzoic Acid, an Antiviral Compound with Activity against Acyclovir-Resistant Isolates of Herpes Simplex Virus Types 1 and 2

2-[4,5-Difluoro-2-(2-Fluorobenzoylamino)-Benzoylamino]Benzoic Acid, an Antiviral Compound with Activity against Acyclovir-Resistant Isolates of Herpes Simplex Virus Types 1 and 2

Comparison of Simplexa HSV 1 & 2 PCR with Culture, Immunofluorescence, and Laboratory-Developed TaqMan PCR for Detection of Herpes Simplex Virus in Swab Specimens

Real-Time PCR Quantification of Genital Shedding of Herpes Simplex Virus (HSV) and Human Immunodeficiency Virus (HIV) in Women Coinfected with HSV and HIV

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