Table 2.

Effect of Buffer RLT treatment on DNA and RNA enrichment from yeast cells for mixtures composed of different ratios of PBMCs and Candida cells, as determined by qPCR-based amplification^a of human DNA and fungal DNA, and by RT-qPCR based amplification^a of human RNA and fungal RNA.

Human DNA	Cq valueb	SD	Log cells/mlc	CV	DNA %c	CV
106 PBMCs + 105 Candida cells
No enrichment	24.39	0.67	6.08	0.03	100.00	0.34
Buffer RLT	32.52*	0.55	3.89	0.04	0.65	0.29
106 PBMCs + 104 Candida cells
No enrichment	24.87	0.13	5.95	0.01	100.00	0.08
Buffer RLT	32.95*	0.65	3.77	0.05	0.67	0.33
Fungal DNA	Cq valueb	SD	Log cells/mlc	CV	DNA %c	CV
106 PBMCs + 105 Candida cells
No enrichment	23.74	0.59	5.02	0.03	100.00	0.32
Buffer RLT	24.31	0.58	4.85	0.03	68.56	0.32
106 PBMCs + 104 Candida cells
No enrichment	27.77	0.10	3.85	0.01	100.00	0.07
Buffer RLT	28.72	0.25	3.57	0.07	53.05	0.45
Human RNA	Cq valueb	SD	Log cells/mlc	CV	RNA %c	CV
106 PBMCs + 105 Candida cells
No enrichment	30.65	0.88	6.09	0.04	100.00	0.45
Buffer RLT	35.15*	0.98	4.78	0.06	4.91	0.48
106 PBMCs + 104 Candida cells
No enrichment	30.74	1.65	4.37	0.10	100.00	0.64
Buffer RLT	34.57*	0.71	3.34	0.06	9.30	0.36
Fungal RNA	Cq valueb	SD	Log cells/mlc	CV	RNA %c	CV
106 PBMCs + 105 Candida cells
No enrichment	22.51	0.52	5.06	0.03	100.00	0.28
Buffer RLT	23.26	0.46	4.84	0.03	61.47	0.26
106 PBMCs + 104 Candida cells
No enrichment	25.47	0.54	4.22	0.04	100.00	0.30
Buffer RLT	27.10	0.82	3.76	0.06	34.69	0.41

^aHuman nucleic acids amplified with CXCL-1 primers and fungal nucleic acids amplified with ITS-1 primers.

^bCq values are means of six biological replicates. Statistically significant differences are marked with one asterisk (p < 0.05) (paired-samples Wilcoxon test).

^cLog cells/ml, DNA percentage and RNA percentage were calculated by extrapolation of Cq values with a standard curve.

SD: Standard deviation. CV: Coefficient of variation.