Abstract
The potential advantage of using fast PCR to detect human herpesvirus 8 (HHV-8) was tested by running a rapid cycling protocol (5. s-steps) in one standard and two fast ramping thermal cyclers to evaluate the performance of 8 different fast reagents. Under this extremely short time profile, assay sensitivity comparable to that of the original protocol was maintained using fast reagents from five suppliers. Reproducibility was higher using fast ramping thermal cyclers, suggesting that fast chemistry may be better matched with advanced instruments. Few fast reagents showed a 2-log-increase in sensitivity and good consistency, that allowed the substitution of the standard nested PCR method with the fast, single round technique. Overall, these studies indicate that some of the fast reagents tested may be used to perform a highly sensitive and reproducible HHV-8 detection with a considerable time saving.
| Original language | English |
|---|---|
| Pages (from-to) | 125-130 |
| Number of pages | 6 |
| Journal | Journal of Virological Methods |
| Volume | 181 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Apr 2012 |
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Keywords
- Fast PCR
- Fast ramping thermal cycler
- HHV-8
- KSHV
ASJC Scopus subject areas
- Virology
Cite this
Evaluation of fast PCR reagents for rapid and sensitive detection of human herpesvirus 8. / Bergamo, Elisa; Chiapolino, Giulia; Lignitto, Laura; Calabrò, Maria Luisa.
In: Journal of Virological Methods, Vol. 181, No. 1, 04.2012, p. 125-130.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Evaluation of fast PCR reagents for rapid and sensitive detection of human herpesvirus 8
AU - Bergamo, Elisa
AU - Chiapolino, Giulia
AU - Lignitto, Laura
AU - Calabrò, Maria Luisa
PY - 2012/4
Y1 - 2012/4
N2 - The potential advantage of using fast PCR to detect human herpesvirus 8 (HHV-8) was tested by running a rapid cycling protocol (5. s-steps) in one standard and two fast ramping thermal cyclers to evaluate the performance of 8 different fast reagents. Under this extremely short time profile, assay sensitivity comparable to that of the original protocol was maintained using fast reagents from five suppliers. Reproducibility was higher using fast ramping thermal cyclers, suggesting that fast chemistry may be better matched with advanced instruments. Few fast reagents showed a 2-log-increase in sensitivity and good consistency, that allowed the substitution of the standard nested PCR method with the fast, single round technique. Overall, these studies indicate that some of the fast reagents tested may be used to perform a highly sensitive and reproducible HHV-8 detection with a considerable time saving.
AB - The potential advantage of using fast PCR to detect human herpesvirus 8 (HHV-8) was tested by running a rapid cycling protocol (5. s-steps) in one standard and two fast ramping thermal cyclers to evaluate the performance of 8 different fast reagents. Under this extremely short time profile, assay sensitivity comparable to that of the original protocol was maintained using fast reagents from five suppliers. Reproducibility was higher using fast ramping thermal cyclers, suggesting that fast chemistry may be better matched with advanced instruments. Few fast reagents showed a 2-log-increase in sensitivity and good consistency, that allowed the substitution of the standard nested PCR method with the fast, single round technique. Overall, these studies indicate that some of the fast reagents tested may be used to perform a highly sensitive and reproducible HHV-8 detection with a considerable time saving.
KW - Fast PCR
KW - Fast ramping thermal cycler
KW - HHV-8
KW - KSHV
UR - http://www.scopus.com/inward/record.url?scp=84858154717&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84858154717&partnerID=8YFLogxK
U2 - 10.1016/j.jviromet.2012.01.002
DO - 10.1016/j.jviromet.2012.01.002
M3 - Article
C2 - 22266419
AN - SCOPUS:84858154717
VL - 181
SP - 125
EP - 130
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 1
ER -