Table 1.
Primers and probes used for HHV-8 and GAPDH qPCR assays
| Assay | Primers/Probe | Sequence (5′-3′) | Nucleotide position* | Amplicon length | Reference |
|---|---|---|---|---|---|
|
Quantitative PCR Assays for HHV-8 | |||||
| HHV-8 ORF73 |
HHV8_73_01.1 |
GGTGATGTTCTGAGTACATAGCGG |
124,326 - 124,349 |
142 bp |
Lallemand et al., 2000 |
| HHV8_73_02.1 |
CCGAGGACGAAATGGAAGTG |
124,467 - 124,448 |
|||
| HHV8_73_Pb1 |
FAM-ACAAATTGCCAGTAGCCCACCAGGAGA-BHQ1ξ |
124,421 - 124,395 |
|||
| HHV-8 ORF26 |
HHV8_26_01.1 |
AGCCGAAAGGATTCCACCATT |
47,287 - 47,304 |
234 bp |
Hammock et al., 2005 |
| HHV8_26_02.1 |
TCCGTGTTGTCTACGTCCAGA |
47,519 - 47,499 |
|||
| HHV8_26_Pb.3 |
FAM-TGCAGCAGYTGTTGGTGTACCACAT-BHQ1ξ |
47,378 - 47,402 |
|||
|
Quantitative PCR Assays for Normalisation to Cell Counts | |||||
| GAPDH |
GAPDH_01.1 |
GCTCCCTCTTTCTTTGCAGCAAT |
7.800 - 7,822 |
104 bp |
Asahi-Ozaki et al., 2006 |
| GAPDH_02.1 |
TACCATGAGTCCTTCCACGATAC |
7,903 - 7,881 |
|||
| GAPDH_Pb1 | FAM-TCCTGCACCACCAACTGCTTAGCACC-BHQ1ξ | 7.826 - 7,851 | |||
*Nucleotide position was determined from the reference sequences [GenBank:NC_003409] for HHV-8 and [GenBank:NG_007073.2] for GAPDH.
ξProbes had 5′ terminus labeled with a FAM (6-carboxy-fluorescein) fluorophore and 3′ terminus labeled with BHQ-1 (Black Hole Quencher®).