Table 1.

The accuracy and sensitivity of a bioluminescent signal-based digital LAMP assay.

Target RNA Concentrations (Copies/mL)	Target RNA Concentrations in Reaction Mix (Copies/μL) (10 μL Reaction Mixtures)	Theoretical Calculation (N = 20,000)			Actual Results
		λ	Np	Nn/N	Np	N	λ	C (Copies/μL)
104	1	0.0005	10	0.9995	9	18,554	0.00049	0.98
105	10	0.0050	99	0.9950	77	17,386	0.00443	8.89
106	100	0.0500	969	0.9516	669	19,097	0.03503	70.06

N: the total number of droplets. Np: number of positive droplets. Nn: number of negative droplets. λ: copies per droplet = −ln(Nn/N) = Copies in the reaction/Total droplets = Concentration in reaction (Copies/μL) × V (volume of each droplet). V = Reaction volume/N = 0.0005 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 = 10⁴³ × C_Copies/mL. P(0) = Nn/N = e^−λ, λ = −ln(Nn/N) = C_Copies/mL × V.