ArticleJournal of clinical microbiology2002
Calibrated real-time PCR assay for quantitation of human herpesvirus 8 DNA in biological fluids.
Article in Journal of clinical microbiology, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- Human Herpes Virus 8 in HIV-1 infected individuals receiving cancer chemotherapy and stem cell transplantation.PloS one · 2018Article
- Calibration technologies for correct determination of Epstein-Barr Virus, human herpesvirus 6 (HHV-6), and HHV-8 antiviral drug susceptibilities by use of real-time-PCR-based assays.Journal of clinical microbiology · 2013Article
- Detection of C. trachomatis in the serum of the patients with urogenital chlamydiosis.BioMed research international · 2013Article
- Isolation of Chlamydia pneumoniae from serum samples of the patients with acute coronary syndrome.International journal of medical sciences · 2010Article
- Chronic human herpes virus 8 infection successfully treated with valganciclovir.The World Allergy Organization journal · 2009Article
- Interactions between human immunodeficiency virus type 1 and vaccinia virus in human lymphoid tissue ex vivo.Journal of virology · 2007Article
- Human herpesvirus 6A accelerates AIDS progression in macaques.Proceedings of the National Academy of Sciences of the United States of America · 2007Article
- Viral interactions in human lymphoid tissue: Human herpesvirus 7 suppresses the replication of CCR5-tropic human immunodeficiency virus type 1 via CD4 modulation.Journal of virology · 2007Article
- Transfer and evaluation of an automated, low-cost real-time reverse transcription-PCR test for diagnosis and monitoring of human immunodeficiency virus type 1 infection in a West African resource-limited setting.Journal of clinical microbiology · 2005Article
- Evidence for both lytic replication and tightly regulated human herpesvirus 8 latency in circulating mononuclear cells, with virus loads frequently below common thresholds of detection.Journal of virology · 2004Article
- Development of a real-time PCR assay using SYBR Green I for provirus load quantification in a murine model of AIDS.Journal of clinical microbiology · 2004Article
- Performance assessment of eight high-throughput PCR assays for viral load quantitation of oncogenic HPV types.The Journal of molecular diagnostics : JMD · 2004Article
- Human herpesvirus 8 load in matched serum and plasma samples of patients with AIDS-associated Kaposi's sarcoma.Journal of clinical microbiology · 2003Article
- Rapid diagnosis of mycobacterial infections and quantitation of Mycobacterium tuberculosis load by two real-time calibrated PCR assays.Journal of clinical microbiology · 2003Article
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Authors and funding
10 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Accurate laboratory tests for the diagnosis of active human herpesvirus 8 (HHV-8) infection are becoming essential to study the pathogenesis of HHV-8-associated tumors and for the clinical management of HHV-8-infected individuals. We have developed a highly sensitive, calibrated quantitative real-time PCR assay for the measurement of cell-free HHV-8 DNA in body fluids, based on the addition of a synthetic DNA calibrator prior to DNA extraction. The calibrator controls each sample for the presence of PCR inhibitors, determines a cutoff value of sensitivity for negative samples, and normalizes positive samples for the efficiency of DNA recovery. The assay shows a wide dynamic range of detection (between 1 and 10(6) viral genome equivalents/reaction) and a high degree of accuracy even in the presence of high amounts (up to 1 micro g) of human genomic DNA. Moreover, the assay has a very high sensitivity (lower detection limit, 10 genome equivalents/ml) and a high degree of reproducibility and repeatability with a coefficient of variation (CV) of <15 and 23%, respectively. Furthermore, the use of the calibrator improves the accuracy of quantitation and decreases the intersample variability (CV, 9 and 6%, respectively). The sensitivity and specificity of the assay were tested with a series of clinical specimens obtained from patients affected by various HHV-8-related diseases, as well as from a wide number of controls. In conclusion, our calibrated real-time PCR assay provides a reliable high-throughput method for quantitation of HHV-8 DNA in clinical and laboratory specimens.
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