Table 3.

The accuracy of the bioluminescent signal-based simulated digital LAMP assay.

Target RNA Concentrations (Copies/mL)	Target RNA Concentrations in Reaction (Copies/μL) (1000 μL Reaction Mixtures)	Theoretical Calculation (N = 96)			Actual Results
		λ	Np	Nn/N	Np	N	λ	C (Copies/μL)
0	0	0	0	1	0	96	0	0
103	0.1	1.0416	62	0.3535	60	96	0.9808	0.0943
104	1	10.4166	96	0	96	96	≥9.6	≥9.6

N: the total number of wells. Np: number of positive wells. Nn: number of negative wells. λ: copies per well = −ln(Nn/N) = Copies in the reaction/Total wells = Concentration in reaction (Copies/μL) × V (volume of each well). V = Reaction volume/N = 10.4 μL. P(0) = Nn/N = e^−λ, λ = −ln(Nn/N) = Concentration in reaction × V. Concentration in reaction = [−ln(N/N)]/V. Target RNA concentration = 10 × C_Copies/μL = 10⁴ × C_Copies/mL.