A Real-Time PCR Assay for Bat SARS-Like Coronavirus Detection and Its Application to Italian Greater Horseshoe Bat Faecal Sample Surveys

Table 2

Intra- and interassay variability of the SYBR green real-time PCR method.

Intra- and interassay variability
Samples	Replicate (and assay) numbers	Mean (SD)	log10 mean (SD)	log10 CV%
Intraassay variability
A1	3 (1)	1,86𝐸 + 06 (2,35𝐸 + 05)	6,27 (0,05)	0,84
A2	3 (1)	2,03𝐸 + 05 (1,77𝐸 + 04)	5,31 (0,04)	0,71
A3	3 (1)	2,00𝐸 + 04 (8,96𝐸 + 02)	4,30 (0,02)	0,45
A4	3 (1)	2,26𝐸 + 03 (2,53𝐸 + 02)	3,35 (0,05)	1,41
A5	3 (1)	2,09𝐸 + 02 (8,22)	2,32 (0,02)	0,74
A6	3 (1)	1,98𝐸 + 01 (5,29)	1,28 (0,13)	9,99
A7	3 (1)	2,09𝐸+ 00 (0,44)	0,31 (0,09)	28,58
B1	3 (1)	5,79𝐸 + 02 (4,65𝐸 + 01)	2,76 (0,03)	1,27
B2	3 (1)	2,00𝐸 + 00 (0,10)	0,30 (0,02)	7,56
B3	3 (1)	4,01𝐸 + 00 (0,74)	0,60 (0,08)	13,68
Interassay variability
B1	3 (3)	6,08𝐸 + 02 (5,58𝐸 + 01)	2,78 (0,04)	1,38
B2	3 (3)	2,06𝐸 + 00 (0,34)	0,29 (0,07)	23,97
B3	3 (3)	3,11𝐸 + 00 (0,82)	0,46 (0,13)	28,79

Successive 10-fold dilutions of recombinant plasmid: A1 (2×10⁶), A2 (2×10⁵), A3 (2×10⁴), A4 (2×10³), A5 (2×10²), A6 (2×10¹), and A7 (2×10⁰).
Three bat samples with different viral concentrations: B1 (10²), B2 and B3 (10⁰).
SD: standard deviation.
CV: coefficient of variation.