scholarly journals Evaluation of Susceptibility of Human Herpesvirus 8 to Antiviral Drugs by Quantitative Real-Time PCR

2003 ◽  
Vol 41 (8) ◽  
pp. 3897-3900 ◽  
Author(s):  
Y. Sergerie ◽  
G. Boivin
Herpesviridae ◽  
2010 ◽  
Vol 1 (1) ◽  
pp. 3 ◽  
Author(s):  
Vivian Kourí ◽  
Pedro A Martínez ◽  
Orestes Blanco ◽  
Virginia Capó ◽  
María E Rodríguez ◽  
...  

2000 ◽  
Vol 38 (4) ◽  
pp. 1404-1408 ◽  
Author(s):  
Francis Lallemand ◽  
Nathalie Desire ◽  
Willy Rozenbaum ◽  
Jean-Claude Nicolas ◽  
Vincent Marechal

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).


2002 ◽  
Vol 40 (12) ◽  
pp. 4652-4658 ◽  
Author(s):  
F. Broccolo ◽  
G. Locatelli ◽  
L. Sarmati ◽  
S. Piergiovanni ◽  
F. Veglia ◽  
...  

2000 ◽  
Vol 38 (5) ◽  
pp. 1992-1995 ◽  
Author(s):  
Irene E. White ◽  
Thomas B. Campbell

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.


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