Calibrated real-time polymerase chain reaction for specific quantitation of HHV-6A and HHV-6B in clinical samples

Giulia Cassina, Domenico Russo, Davide De Battista, Francesco Broccolo, Paolo Lusso, Mauro S. Malnati

Research output: Contribution to journalArticle

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 languageEnglish
Pages (from-to)172-179
Number of pages8
JournalJournal of Virological Methods
Volume189
Issue number1
DOIs
Publication statusPublished - Apr 2013

Fingerprint

Human Herpesvirus 6
Real-Time Polymerase Chain Reaction
Viruses
Herpesviridae
DNA
Viral DNA
Coinfection
Technology

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 journalArticle

Cassina, Giulia ; Russo, Domenico ; De Battista, Davide ; Broccolo, Francesco ; Lusso, Paolo ; Malnati, Mauro S. / Calibrated real-time polymerase chain reaction for specific quantitation of HHV-6A and HHV-6B in clinical samples. In: Journal of Virological Methods. 2013 ; Vol. 189, No. 1. pp. 172-179.
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