title: Evaluation of fast PCR reagents for rapid and sensitive detection of human herpesvirus 8.
abstract: The potential advantage of using fast PCR to detect human herpesvirus 8 (HHV-8) was tested by running a rapid cycling protocol (5s-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.
authors.affiliation: Immunology and Diagnostic Molecular Oncology, Istituto Oncologico Veneto, IOV-IRCCS, Padova, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Bergamo
authors.forename: Giulia
authors.initials: G
authors.lastname: Chiapolino
authors.forename: Laura
authors.initials: L
authors.lastname: Lignitto
authors.forename: Maria Luisa
authors.initials: ML
authors.lastname: Calabrò
mesh_terms: D006566:Herpesviridae Infections / Q000175:diagnosis* / Q000821:virology; D019288:Herpesvirus 8, Human / Q000302:isolation & purification*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors; D014773:Virology / Q000379:methods*
chemical_list: D011933:Reagent Kits, Diagnostic