A multiplex real-time PCR quantitation of human herpesvirus-6, 7, 8 viruses: application in blood transfusions

Table 2 Optimization of PCR conditions

Type of ingredient	Gradient (variation rate)		Optimal ultimate concentration (type of virus)
Mgcl₂^a	2.5 mM–5.5 mM (0.5)	3.5 mM (HHV-6)	4 mM (HHV-7)	3.5 mM (HHV-8)
Deoxynucleotide mixture^a	0.14 mM–0.32 mM (0.02)	0.16 mM (HHV-6)	0.16 mM (HHV-7)	0.16 mM (HHV-8)
Primer^a	0.26 μM–0.35 μM (0.01)	0.29 μM (HHV-6)	0.29 μM (HHV-7)	0.31 μM (HHV-8)
Probe^a	0.12 μM–0.40 (0.02)	0.24 μM (HHV-6)	0.26 μM (HHV-7)	0.24 μM (HHV-8)
Taq DNA polymerase^a,b	0.1U–0.6U (0.1)	0.5U (HHV-6)	0.6U (HHV-7)	0.6U (HHV-8)

^aInitial concentration of Mgcl₂, deoxynucleotide mixturea, primer, probe, Taq DNA Polymerase is 25 mM, 10 mM, 10 μM, 10 μM, 5U/μL respectively
^bTaq DNA Polymerase was obtained from TaKaRa Biotechnology Dalian Co. Ltd., Liaoning, China

ISSN: 1743-422X