scholarly journals Calibrated Real-Time PCR Assay for Quantitation of Human Herpesvirus 8 DNA in Biological Fluids

2002 ◽  
Vol 40 (12) ◽  
pp. 4652-4658 ◽  
Author(s):  
F. Broccolo ◽  
G. Locatelli ◽  
L. Sarmati ◽  
S. Piergiovanni ◽  
F. Veglia ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Biological Fluids ◽  
Human Herpesvirus ◽  
Human Herpesvirus 8 ◽  
Pcr Assay ◽  
Herpesvirus 8

scholarly journals Simultaneous quantification of human herpesvirus 8 DNA by real time PCR in different tissues of HIV infected cuban patients with Kaposi's sarcoma

Herpesviridae ◽  
2010 ◽  
Vol 1 (1) ◽  
pp. 3 ◽  
Author(s):  
Vivian Kourí ◽  
Pedro A Martínez ◽  
Orestes Blanco ◽  
Virginia Capó ◽  
María E Rodríguez ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Kaposi's Sarcoma ◽  
Human Herpesvirus ◽  
Kaposi’S Sarcoma ◽  
Human Herpesvirus 8 ◽  
Herpesvirus 8 ◽  
Simultaneous Quantification ◽  
Different Tissues

scholarly journals Quantitative Analysis of Human Herpesvirus 8 Viral Load Using a Real-Time PCR Assay

2000 ◽  
Vol 38 (4) ◽  
pp. 1404-1408 ◽  
Author(s):  
Francis Lallemand ◽  
Nathalie Desire ◽  
Willy Rozenbaum ◽  
Jean-Claude Nicolas ◽  
Vincent Marechal
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Clinical Symptoms ◽  
Human Herpesvirus ◽  
Human Herpesvirus 8 ◽  
Clinical Samples ◽  
Taqman Assay ◽  
Published Data ◽  
Herpesvirus 8 ◽  
Viral Loads

We have developed a quantitative real-time PCR (TaqMan) assay aimed at measuring the cellular human herpesvirus 8 (HHV-8) DNA load in various clinical samples. Standard curves were obtained by serial dilutions of a control plasmid containing both HHV-8 (ORF73 gene) and the cellular target (human albumin gene). The assay appeared to be very sensitive (100% detection rate for at least 10 copies per well) and specific and was easily reproducible (less than 3% intra-assay variability, 5% interassay variability). This method allowed us to quantify precisely the average HHV-8 copy number per cell in various persistently HHV-8-infected cell lines (BBG-1 cells, n= 200; BC-1 cells, n = 59; BCBL-1 cells,n = 70). A retrospective study was also conducted to assess the HHV-8 DNA load in 12 human immunodeficiency virus-infected patients with either Kaposi's sarcoma (KS; seven patients monitored over a 3-month period) or multicentric Castleman's disease (MCD; five patients). The HHV-8 DNA load ranged from 0 to 9,171 copies/106 cells in low-risk KS patients (T0, I0, S0 according to the classification of the AIDS Clinical Trials group). We also measured the viral loads in MCD patients either during symptomatic periods or during remission. The results are in agreement with previously published data, with high viral loads correlating with clinical symptoms (1.3 × 106 copies/106cells) and low viral loads correlating with asymptomatic periods (less than 5,000 copies/106 cells).

scholarly journals Quantitation of Cell-Free and Cell-Associated Kaposi's Sarcoma-Associated Herpesvirus DNA by Real-Time PCR

2000 ◽  
Vol 38 (5) ◽  
pp. 1992-1995 ◽  
Author(s):  
Irene E. White ◽  
Thomas B. Campbell
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Kaposi's Sarcoma ◽  
Dynamic Range ◽  
Human Herpesvirus ◽  
Kaposi’S Sarcoma ◽  
Linear Dynamic Range ◽  
Pcr Assay ◽  
Herpesvirus 8 ◽  
Linear Dynamic

A real-time PCR assay for quantitation of Kaposi's sarcoma-associated herpes virus (KSHV or human herpesvirus 8) DNA was evaluated. The linear dynamic range was 10 to 105 copies of KSHV DNA (r 2 > 0.99). The accuracy of DNA purification and quantitation was less than ±0.4 log10copies for samples that contained from 10 to 105 copies of KSHV DNA. Cell-associated KSHV DNA was quantitated over a range of infected cell frequencies from 0.1 to 10−5, and cell-free KSHV DNA in plasma was quantitated over a range of 100 to 106 copies/ml. Real-time PCR provides a convenient method for quantitation of cell-free and cell-associated KSHV DNA in laboratory and clinical specimens.

Development of a LUX real-time PCR for the detection and quantification of human herpesvirus 7

2009 ◽  
Vol 55 (3) ◽  
pp. 319-325 ◽  
Author(s):  
Massimiliano Bergallo ◽  
Cristina Costa ◽  
Maria Elena Terlizzi ◽  
Francesca Sidoti ◽  
Samuela Margio ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Primary Infection ◽  
Latent Infection ◽  
Dynamic Range ◽  
Human Herpesvirus ◽  
Clinical Samples ◽  
House Light ◽  
Pcr Assay ◽  
Sensitivity Specificity

Human herpesvirus 7 is a highly seroprevalent β-herpesvirus that, following primary infection, remains latent in CD4+ T cells and determines a persistent rather than a latent infection in various tissues and organs, including the lung and skin. This paper describes the development of an in-house light upon extension real-time PCR assay for quantification of human herpesvirus 7 DNA in clinical samples. The efficiency, sensitivity, specificity, inter- and intra-assay variability, and dynamic range have been determined. Subsequently, the assay has been validated by evaluating the human herpesvirus 7 load in bronchoalveolar lavages and skin specimens, chosen as 2 persistency sites, from healthy and pathological individuals. The real-time PCR assay developed in this study could be a useful tool to detect and quantify human herpesvirus 7 DNA in different clinical specimens to elucidate its epidemiological and pathogenic roles.

Sign in / Sign up

Export Citation Format

Share Document