Development and evaluation of a TaqMan MGB RT-PCR assay for detection of H5 and N8 subtype influenza virus

BMC Infectious Diseases

Table 3 Intra-assay variation in multiplex detection of H5 and N8 avian influenza viruses

Plasmid copy number (per reaction)	Gene	Ct value in intra-assays			Mean ± SD	CV%
Plasmid copy number (per reaction)	Gene	1	2	3	Mean ± SD	CV%
1 × 10⁵	H5	24.36	24.37	24.30	24.34 ± 0.03	0.13%
1 × 10⁵	N8	23.97	23.72	23.97	23.89 ± 0.12	0.49%
1 × 10⁴	H5	28.73	28.88	29.07	28.89 ± 0.14	0.48%
1 × 10⁴	N8	26.97	27.04	27.02	27.01 ± 0.03	0.11%
1 × 10³	H5	32.01	32.12	32.00	32.04 ± 0.05	0.17%
1 × 10³	N8	30.01	30.23	30.14	30.13 ± 0.09	0.30%
1 × 10²	H5	34.79	34.51	34.68	34.66 ± 0.12	0.33%
1 × 10²	N8	34.05	34.44	34.41	34.30 ± 0.18	0.52%
1 × 10¹	H5	38.59	38.23	38.13	38.32 ± 0.20	0.52%
1 × 10¹	N8	37.86	37.77	37.87	37.83 ± 0.04	0.12%
1 × 10⁰	H5	No Ct	No Ct	No Ct
1 × 10⁰	N8	No Ct	No Ct	No Ct

Note: Each concentration included three replicates on one plate. CV% (threshold = 3%) and Ct values for each concentration are shown

ISSN: 1471-2334