Rapid quantification of hepatitis B virus DNA by real-time PCR using efficient TaqMan probe and extraction of virus DNA

Table 1 Copy number and Ct values of HBV serum samples derived from three different standard curves (mean  ±  SD)

Standardcurve	Ct		Copy number ( /mL)
	Mean ± SD	CV(%)	Mean ± SD	CV (%)
S	26.6 ± 0.061	0.2	5.7 × 104± 2.29 × 103	4.0
C	31.8 ± 0.073	0.2	6.3 × 102± 2.73 × 101	4.3
X	29.5 ± 0.357	1.2	1.6 × 103± 1.77 × 102	11.1

CV: Coefficient variation.

Table 2 Copy number and Ct values of HBV serum samples detected by six different extraction methods (mean ± SD)

Method	Copy number/mL and Ct value
	Sample A	Sample B	Sample C
Guanidinium isothiocyanate	1.41 × 105± 1.23 × 104	1.92 × 105± 1.70 × 104	3.41 × 104± 3.65 × 103
	31.3 ± 0.16	30.9 ± 0.07	33.3 ± 1.59
Proteinase K	2.99 × 105± 2.88 × 104	2.21 × 105± 2.97 × 104	3.59 × 105± 5.10 × 104
	30.5 ± 0.19	30.3 ± 0.38	29.6 ± 0.38
NaI	2.51 × 105± 2.91 × 104	2.12 × 105± 3.01 × 104	9.12 × 104± 9.65 × 103
	30.7 ± 0.17	30.5 ± 0.66	31.4 ± 0.14
NaOH lysis	3.49 × 108± 2.72 × 107	2.08 × 106± 2.80 × 105	4.40 × 107± 2.06 × 106
	19.9 ± 0.18	30.0 ± 0.52	26.2 ± 0.78
Alkaline lysis	N/ N	N/ N	N/ N
Boiling	2.14 × 108± 2.16 × 107	1.83 × 106± 1.05 × 105	1.48 × 107± 1.22 × 106
	23.9 ± 0.10	30.1 ± 0.69	27.4 ± 0.94

N: Negative.

Table 3 Copy number and Ct values of 8 clinical HBV serum samples detected by optimized primers, TaqMan probe from S region and NaOH lysis method (mean ± SD)

Sample	Ct		Copy number /mL
	Mean ± SD	CV (%)	Mean ± SD	CV (%)
1	25.4 ± 0.24	0.9	1.4 × 109± 2.38 × 108	17.0
2	31.9 ± 0.20	0.6	1.4 × 107± 1.87 × 106	13.4
3	31.5 ± 0.52	1.7	1.8 × 107± 2.32 × 106	12.9
4	27.3 ± 0.14	0.5	3.7 × 108± 3.73 × 107	10.0
5	34.0 ± 0.27	8.1	6.4 × 106± 6.08 × 105	9.5
6	32.3 ± 0.52	0.2	1.1 × 107± 4.43 × 106	4.0
7	27.0 ± 0.23	0.8	4.6 × 108± 7.22 × 106	15.7
8	33.9 ± 0.12	0.4	3.1 × 106± 2.76 × 105	8.9

CV: Coefficient variation.