title: Calibrated real-time polymerase chain reaction for specific quantitation of HHV-6A and HHV-6B in clinical samples.
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 10(0) to 10(6) 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.
authors: Unit of Human Virology Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, Via Olgettina 58, 20132 Milan, Italy.
authors: Giulia
authors: G
authors: Cassina
authors: Domenico
authors: D
authors: Russo
authors: Davide
authors: D
authors: De Battista
authors: Francesco
authors: F
authors: Broccolo
authors: Paolo
authors: P
authors: Lusso
authors: Mauro S
authors: MS
authors: Malnati
mesh_terms: D001483:Base Sequence; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D015654:Herpesvirus 6, Human / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D019349:Roseolovirus Infections / Q000175:diagnosis*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D019562:Viral Load
chemical_list: D004279:DNA, Viral