Abstract
The recent classification of human herpesvirus 6 (HHV-6) A and B, previously considered as two variants of the same virus, as two distinct herpesvirus species, emphasizes the need to develop and standardize specific methods for their detection and quantitation for clinical use. The development of two highly sensitive calibrated real-time PCR to quantify HHV-6A and -6B variants in clinical specimen is described. Both assays displayed the same wide linear dynamic range from 100 to 106 copies of viral DNA in a single reaction and sensitivity of one copy/reaction. These systems allow for HHV-6A/B DNA load quantitation in different types of clinical specimens: blood or tissue cells when combined with the CCR5 assay; cell-free samples (plasma or other biological fluids) in combination with the calibrator technology. Due to the absence of cross-amplification and cross-hybridization, these methods detect minute amounts of one viral species even in the presence of a large excess of the other, allowing a specific quantitation of both viruses in the case of mixed infections. The new qPCR methods provide sensitive and specific tool for monitoring HHV-6A/B DNA load in clinical samples, facilitating the study of these viruses in human diseases.
| Original language | English |
|---|---|
| Pages (from-to) | 172-179 |
| Number of pages | 8 |
| Journal | Journal of Virological Methods |
| Volume | 189 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Apr 2013 |
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Keywords
- Calibrated PCR
- Clinical diagnosis
- DNA load
- HHV-6A
- HHV-6B
- Real-time quantitative PCR
ASJC Scopus subject areas
- Virology
Cite this
Calibrated real-time polymerase chain reaction for specific quantitation of HHV-6A and HHV-6B in clinical samples. / Cassina, Giulia; Russo, Domenico; De Battista, Davide; Broccolo, Francesco; Lusso, Paolo; Malnati, Mauro S.
In: Journal of Virological Methods, Vol. 189, No. 1, 04.2013, p. 172-179.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Calibrated real-time polymerase chain reaction for specific quantitation of HHV-6A and HHV-6B in clinical samples
AU - Cassina, Giulia
AU - Russo, Domenico
AU - De Battista, Davide
AU - Broccolo, Francesco
AU - Lusso, Paolo
AU - Malnati, Mauro S.
PY - 2013/4
Y1 - 2013/4
N2 - The recent classification of human herpesvirus 6 (HHV-6) A and B, previously considered as two variants of the same virus, as two distinct herpesvirus species, emphasizes the need to develop and standardize specific methods for their detection and quantitation for clinical use. The development of two highly sensitive calibrated real-time PCR to quantify HHV-6A and -6B variants in clinical specimen is described. Both assays displayed the same wide linear dynamic range from 100 to 106 copies of viral DNA in a single reaction and sensitivity of one copy/reaction. These systems allow for HHV-6A/B DNA load quantitation in different types of clinical specimens: blood or tissue cells when combined with the CCR5 assay; cell-free samples (plasma or other biological fluids) in combination with the calibrator technology. Due to the absence of cross-amplification and cross-hybridization, these methods detect minute amounts of one viral species even in the presence of a large excess of the other, allowing a specific quantitation of both viruses in the case of mixed infections. The new qPCR methods provide sensitive and specific tool for monitoring HHV-6A/B DNA load in clinical samples, facilitating the study of these viruses in human diseases.
AB - The recent classification of human herpesvirus 6 (HHV-6) A and B, previously considered as two variants of the same virus, as two distinct herpesvirus species, emphasizes the need to develop and standardize specific methods for their detection and quantitation for clinical use. The development of two highly sensitive calibrated real-time PCR to quantify HHV-6A and -6B variants in clinical specimen is described. Both assays displayed the same wide linear dynamic range from 100 to 106 copies of viral DNA in a single reaction and sensitivity of one copy/reaction. These systems allow for HHV-6A/B DNA load quantitation in different types of clinical specimens: blood or tissue cells when combined with the CCR5 assay; cell-free samples (plasma or other biological fluids) in combination with the calibrator technology. Due to the absence of cross-amplification and cross-hybridization, these methods detect minute amounts of one viral species even in the presence of a large excess of the other, allowing a specific quantitation of both viruses in the case of mixed infections. The new qPCR methods provide sensitive and specific tool for monitoring HHV-6A/B DNA load in clinical samples, facilitating the study of these viruses in human diseases.
KW - Calibrated PCR
KW - Clinical diagnosis
KW - DNA load
KW - HHV-6A
KW - HHV-6B
KW - Real-time quantitative PCR
UR - http://www.scopus.com/inward/record.url?scp=84875515600&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84875515600&partnerID=8YFLogxK
U2 - 10.1016/j.jviromet.2013.01.018
DO - 10.1016/j.jviromet.2013.01.018
M3 - Article
C2 - 23391825
AN - SCOPUS:84875515600
VL - 189
SP - 172
EP - 179
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 1
ER -