Identification and validation of suitable endogenous reference genes for gene expression studies in human peripheral blood

Table 5 Diagnosis effect on the candidate control genes.

	ΔΔCt Method	Comparative Ct Method
	Normalized to TRAP1	Normalized to Optimal Set of 4 Genes*
Gene	Fold Change (MS/H)	Fold Change (MS/H)	p-value(Diagnosis)
FPGS	1.08	1.00	0.964
ACTb	1.08	ND	0.788
GINS2	1.21	1.11	0.691
HMBS	1.12	1.06	0.581
B2M	-1.04	-1.10	0.575
FARP1	1.34	1.16	0.559
A4GALT	-1.01	ND	0.416
TRAP1	**	-1.08	0.342
GAPDH	-1.45	-1.61	0.198
DECR1	-1.09	-1.11	0.161
PPIB	1.32	1.22	0.155
PEX16	1.32	1.23	0.073
MAPRE2	1.42	1.30	0.072
CRY2	1.58	1.45	0.065
CSNK1G2	1.76	1.63	0.026

Expression of the candidate reference genes in whole blood of multiple sclerosis (MS) subjects is compared to age and gender matched control subjects using the ΔΔCt Method and the Comparative Ct Method. Genes are arranged in the order of decreasing p-value for diagnosis.
*ANOVA: μ + Diagnosis + Plate + (Diagnosis × Plate) + ε
**TRAP1 fold change not calculated since that gene was used as a normalizer.

ISSN: 1755-8794