Viral loads of herpes simplex virus in clinical samples-A 5-year retrospective analysis

2010 ◽  
Vol 82 (11) ◽  
pp. 1911-1916 ◽  
Author(s):  
Julian W. Tang ◽  
Mingxuan Lin ◽  
Lily Chiu ◽  
Evelyn S.C. Koay
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Retrospective Analysis ◽  
Clinical Samples ◽  
Viral Loads ◽  
Simplex Virus

Differentiation of herpes simplex virus types 1 and 2 in clinical samples by a real-time taqman PCR assay

2005 ◽  
Vol 76 (3) ◽  
pp. 350-355 ◽  
Author(s):  
Lawrence Corey ◽  
Meei-Li Huang ◽  
Stacy Selke ◽  
Anna Wald
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Real Time ◽  
Clinical Samples ◽  
Pcr Assay ◽  
Taqman Pcr ◽  
Simplex Virus

scholarly journals Development of a High-Throughput Quantitative Assay for Detecting Herpes Simplex Virus DNA in Clinical Samples

1999 ◽  
Vol 37 (6) ◽  
pp. 1941-1947 ◽  
Author(s):  
Alexander J. Ryncarz ◽  
James Goddard ◽  
Anna Wald ◽  
Meei-Li Huang ◽  
Bernard Roizman ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Real Time ◽  
High Throughput ◽  
Real Time Pcr ◽  
Genital Tract ◽  
Clinical Samples ◽  
Pcr Assay ◽  
The Mean ◽  
Simplex Virus

We have developed a high-throughput, semiautomated, quantitative fluorescence-based PCR assay to detect and type herpes simplex virus (HSV) DNA in clinical samples. The detection assay, which uses primers to the type-common region of HSV glycoprotein B (gB), was linear from <10 to 108 copies of HSV DNA/20 μl of sample. Among duplicate samples in reproducibility runs, the assay showed less than 5% variability. We compared the fluorescence-based PCR assay with culture and gel-based liquid hybridization system with 335 genital tract specimens from HSV type 2 (HSV-2)-seropositive persons attending a research clinic and 380 consecutive cerebrospinal fluid (CSF) samples submitted to a diagnostic virology laboratory. Among the 162 culture-positive genital tract specimens, TaqMan PCR was positive for 157 (97%) specimens, whereas the quantitative-competitive PCR was positive for 144 (89%) specimens. Comparisons of the mean titer of HSV DNA detected by the two assays revealed that the mean titer detected by the gel-based system was slightly higher (median, 1 log). These differences in titers were in part related to the fivefold difference in the amount of HSV DNA used in the amplicon standards with the two assays. Among the 380 CSF samples, 42 were positive by both assays, 13 were positive only by the assay with the agarose gel, and 3 were positive only by the assay with the fluorescent probe. To define the subtype of HSV DNA detected in the screening assay, we also designed one set of primers which amplifies the gG regions of both types of HSV and probes which are specific to either HSV-1 (gG1) or HSV-2 (gG2). These probes were labeled with different fluorescent dyes (6-carboxyfluorescein for gG2 and 6-hexachlorofluorescein for gG1) to enable detection in a single PCR. In mixing experiments the probes discriminated the correct subtype in mixtures with up to a 7-log-higher concentration of the opposite subtype. The PCR typing results showed 100% concordance with the results obtained by assays with monoclonal antibodies against HSV-1 or HSV-2. Thus, while the real-time PCR is slightly less sensitive than the gel-based liquid hybridization system, the high throughput, the lack of contamination during processing, the better reproducibility, and the better ability to type the isolates rapidly make the real-time PCR a valuable tool for clinical investigation and diagnosis of HSV infection.

scholarly journals Herpes simplex virus 2 serostatus and viral loads of HIV-1 in blood and semen as risk factors for HIV transmission among men who have sex with men

AIDS ◽  
2008 ◽  
Vol 22 (13) ◽  
pp. 1667-1671 ◽  
Author(s):  
David M Butler ◽  
Davey M Smith ◽  
Edward R Cachay ◽  
George K Hightower ◽  
Charles Thomas Nugent ◽  
...  
Keyword(s):  
Risk Factors ◽  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Hiv Transmission ◽  
Viral Loads ◽  
Sex With Men ◽  
Simplex Virus ◽  
Herpes Simplex Virus 2 ◽  
Hiv 1

Occurrence of herpes simplex virus 1 and three periodontal bacteria in patients with chronic periodontitis and necrotic pulp

2008 ◽  
Vol 54 (4) ◽  
pp. 326-330 ◽  
Author(s):  
Sheila Alexandra Belini Nishiyama ◽  
Viviane Nakano ◽  
Gustavo Velásquez-Melendez ◽  
Mario Julio Avila-Campos
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Chronic Periodontitis ◽  
Clinical Samples ◽  
Treponema Denticola ◽  
Tannerella Forsythia ◽  
Herpes Simplex Virus 1 ◽  
Nested Polymerase Chain Reaction ◽  
Periodontal Bacteria ◽  
Simplex Virus

Viral and bacterial associations appear to be implicated in the development of periodontal infections. Little information is available describing the periodontopathic agents in root canals with necrotic pulp. In this study, the occurrence and the combinations among herpes simplex virus type 1 (HSV-1) and Dialister pneumosintes , Tannerella forsythia , and Treponema denticola in patients with chronic periodontitis and necrotic pulp were evaluated. Clinical samples from healthy subjects and patients with periodontal or pulp infections were analyzed using a nested polymerase chain reaction PCR to detect HSV and PCR to detect the 3 periodontal bacteria. The presence of Tannerella forsythia and Treponema denticola was observed in healthy, periodontitis, and necrotic pulp patients. HSV was observed in periodontitis and necrotic pulp patients, and no healthy subject harbored D. pneumosintes or HSV. The occurrence of Tannerella forsythia was not statistically significant in patients with necrotic pulp (P = 0.704). Periodontal bacteria were observed varying from 10.3% to 20.7% in periodontitis and necrotic pulp patients. The presence of Treponema denticola – HSV association was predominant in patients showing necrotic pulp (24.1%); however, HSV alone was observed in one patient with periodontitis and in another patient with necrotic pulp. The presence of double association among bacteria or bacteria – HSV could indicate a role in both periodontitis and necrotic pulp, and Tannerella forsythia – Treponema denticola – HSV and Tannerella forsythia – D. pneumosintes – Treponema denticola – HSV associations might be important in periodontitis.

scholarly journals Unlabeled Probes for the Detection and Typing of Herpes Simplex Virus

2007 ◽  
Vol 53 (10) ◽  
pp. 1847-1854 ◽  
Author(s):  
Shale Dames ◽  
David C Pattison ◽  
L Kathryn Bromley ◽  
Carl T Wittwer ◽  
Karl V Voelkerding
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Internal Control ◽  
Clinical Samples ◽  
Attractive Alternative ◽  
Nucleotide Polymorphisms ◽  
Unlabeled Probe ◽  
Melting Analysis ◽  
Simplex Virus ◽  
Labeled Probe

Abstract Background: Unlabeled probe detection with a double-stranded DNA (dsDNA) binding dye is one method to detect and confirm target amplification after PCR. Unlabeled probes and amplicon melting have been used to detect small deletions and single-nucleotide polymorphisms in assays where template is in abundance. Unlabeled probes have not been applied to low-level target detection, however. Methods: Herpes simplex virus (HSV) was chosen as a model to compare the unlabeled probe method to an in-house reference assay using dual-labeled, minor groove binding probes. A saturating dsDNA dye (LCGreen® Plus) was used for real-time PCR. HSV-1, HSV-2, and an internal control were differentiated by PCR amplicon and unlabeled probe melting analysis after PCR. Results: The unlabeled probe technique displayed 98% concordance with the reference assay for the detection of HSV from a variety of archived clinical samples (n = 182). HSV typing using unlabeled probes was 99% concordant (n = 104) to sequenced clinical samples and allowed for the detection of sequence polymorphisms in the amplicon and under the probe. Conclusions: Unlabeled probes and amplicon melting can be used to detect and genotype as few as 10 copies of target per reaction, restricted only by stochastic limitations. The use of unlabeled probes provides an attractive alternative to conventional fluorescence-labeled, probe-based assays for genotyping and detection of HSV and might be useful for other low-copy targets where typing is informative.

Analysis of varicella-zoster virus and herpes simplex virus in various clinical samples by the use of different PCR assays

2009 ◽  
Vol 160 (1-2) ◽  
pp. 193-196 ◽  
Author(s):  
Niklas Finnström ◽  
Karin Bergsten ◽  
Helena Ström ◽  
Therese Sundell ◽  
Sophie Martin ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Varicella Zoster Virus ◽  
Clinical Samples ◽  
Varicella Zoster ◽  
Pcr Assays ◽  
Simplex Virus

scholarly journals Microglia Reduce Herpes Simplex Virus 1 Lethality of Mice with Decreased T Cell and Interferon Responses in Brains

2021 ◽  
Vol 22 (22) ◽  
pp. 12457
Author(s):  
Meng-Shan Tsai ◽  
Li-Chiu Wang ◽  
Hsien-Yang Tsai ◽  
Yu-Jheng Lin ◽  
Hua-Lin Wu ◽  
...  
Keyword(s):  
T Cells ◽  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Infection Model ◽  
Virus Infectivity ◽  
Herpes Simplex Virus 1 ◽  
Viral Loads ◽  
Simplex Virus ◽  
Hsv 1

Herpes simplex virus 1 (HSV-1) infects the majority of the human population and can induce encephalitis, which is the most common cause of sporadic, fatal encephalitis. An increase of microglia is detected in the brains of encephalitis patients. The issues regarding whether and how microglia protect the host and neurons from HSV-1 infection remain elusive. Using a murine infection model, we showed that HSV-1 infection on corneas increased the number of microglia to outnumber those of infiltrating leukocytes (macrophages, neutrophils, and T cells) and enhanced microglia activation in brains. HSV-1 antigens were detected in brain neurons, which were surrounded by microglia. Microglia depletion increased HSV-1 lethality of mice with elevated brain levels of viral loads, infected neurons, neuron loss, CD4 T cells, CD8 T cells, neutrophils, interferon (IFN)-β, and IFN-γ. In vitro studies demonstrated that microglia from infected mice reduced virus infectivity. Moreover, microglia induced IFN-β and the signaling pathway of signal transducer and activator of transcription (STAT) 1 to inhibit viral replication and damage of neurons. Our study reveals how microglia protect the host and neurons from HSV-1 infection.

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