Evaluation of fast PCR reagents for rapid and sensitive detection of human herpesvirus 8

Elisa Bergamo, Giulia Chiapolino, Laura Lignitto, Maria Luisa Calabrò

Research output: Contribution to journalArticle

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 languageEnglish
Pages (from-to)125-130
Number of pages6
JournalJournal of Virological Methods
Volume181
Issue number1
DOIs
Publication statusPublished - Apr 2012

Keywords

  • Fast PCR
  • Fast ramping thermal cycler
  • HHV-8
  • KSHV

ASJC Scopus subject areas

  • Virology

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