Detection of human herpesvirus 8 by quantitative polymerase chain reaction: development and standardisation of methods

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-BHQ1^ξ	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-BHQ1^ξ	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	GAPDH_02.1	TACCATGAGTCCTTCCACGATAC	7,903 - 7,881	104 bp	Asahi-Ozaki et al., 2006
	GAPDH_Pb1	FAM-TCCTGCACCACCAACTGCTTAGCACC-BHQ1^ξ	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^®).

ISSN: 1471-2334