Patent ID: 8012719

Claim:
A method for the quantitative detection of a target nucleic acid from HHV-6 subtype variants A and/or B in a sample, which comprises the following steps: a) extraction of the nucleic acid from the sample with a calibrator previously added to the sample itself, wherein said calibrator is a nucleic acid containing two regions annealing to the primers SEQ ID NOS: 1 and 2, a third region annealing to the probe SEQ ID NO: 7 that is located in between said two regions and two spacers containing from 1 to 30 nucleotides located between the primer- and probe-annealing regions, respectively, wherein said spacers are mismatched with both HHV-6 A and B nucleic acid sequences; b) mixing the extracted target nucleic acid and calibrator with: i) the following primers pairs: a) SEQ ID NOs: 1 and 2 for the calibrator, and b) SEQ ID NOs: 3 and 4 for HHV-6 type A, and c) SEQ ID NOs: 5 and 6 for HHV-6 type B; ii) the following probes: SEQ ID NO:7 for the calibrator, SEQ ID NO:8 for HHV-6A and SEQ ID NO:9 for HHV-6B, wherein said probes carry a fluorescent reporter label able to generate a fluorescence signal and a quencher label at their 5′ and 3′ ends, respectively; iii) a nucleic acid polymerase with 5′-3′ nuclease activity; c) performing a polymerization reaction whereby a signal associated with the reporters label is detected; and d) quantification of the copy number of the extracted target nucleic acid and of the calibrator copy number recovered upon extraction.