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 106 viral genome equivalents/reaction) and a high degree of accuracy even in the presence of high amounts (up to 1 μg) 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
| Original language | English |
|---|---|
| Pages (from-to) | 4652-4658 |
| Number of pages | 7 |
| Journal | Journal of Clinical Microbiology |
| Volume | 40 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published - Dec 1 2002 |
Fingerprint
ASJC Scopus subject areas
- Microbiology (medical)
- Microbiology
Cite this
Calibrated real-time PCR assay for quantitation of human herpesvirus 8 DNA in biological fluids. / Broccolo, Francesco; Locatelli, Giuseppe; Sarmati, Loredana; Piergiovanni, Sara; Veglia, Fabrizio; Andreoni, Massimo; Buttò, Stefano; Ensoli, Barbara; Lusso, Paolo; Malnati, Mauro S.
In: Journal of Clinical Microbiology, Vol. 40, No. 12, 01.12.2002, p. 4652-4658.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Calibrated real-time PCR assay for quantitation of human herpesvirus 8 DNA in biological fluids
AU - Broccolo, Francesco
AU - Locatelli, Giuseppe
AU - Sarmati, Loredana
AU - Piergiovanni, Sara
AU - Veglia, Fabrizio
AU - Andreoni, Massimo
AU - Buttò, Stefano
AU - Ensoli, Barbara
AU - Lusso, Paolo
AU - Malnati, Mauro S.
PY - 2002/12/1
Y1 - 2002/12/1
N2 - 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 106 viral genome equivalents/reaction) and a high degree of accuracy even in the presence of high amounts (up to 1 μg) 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
AB - 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 106 viral genome equivalents/reaction) and a high degree of accuracy even in the presence of high amounts (up to 1 μg) 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
UR - http://www.scopus.com/inward/record.url?scp=18744393373&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=18744393373&partnerID=8YFLogxK
U2 - 10.1128/JCM.40.12.4652-4658.2002
DO - 10.1128/JCM.40.12.4652-4658.2002
M3 - Article
C2 - 12454167
AN - SCOPUS:18744393373
VL - 40
SP - 4652
EP - 4658
JO - Journal of Clinical Microbiology
JF - Journal of Clinical Microbiology
SN - 0095-1137
IS - 12
ER -