Two nanobody-based immunoassays to differentiate antibodies against genotype 1 and 2 porcine reproductive and respiratory syndrome virus

Table 3 Optimized incubation time of the mixture containing pig sera and nanobody-HRP fusion proteins with the antigen and color reaction times using a checkerboard assay for the g1-2-cELISA and g1-cELISA

Color reaction time (min)	Sera type	Incubation time (min) of antigens, sera and Nbs-HRP fusion proteins
		g1-2-PRRSV-Nb3-HRP			g1-PRRSV-Nb136-HRP
		20	30	40	20	30	40
10	Negative	0.229	1.002	1.089	0.187	0.862	0.989
	Positive	0.174	0.211	0.219	0.114	0.241	0.263
	P/N	0.760	0.211	0.201	0.610	0.280	0.266
15	Negative	0.298	1.108	1.105	0.195	0.942	1.005
	Positive	0.172	0.198	0.207	0.115	0.243	0.267
	P/N	0.577	0.179	0.187	0.590	0.258	0.266
20	Negative	0.299	1.112	1.18	0.209	0.978	1.012
	Positive	0.177	0.212	0.221	0.127	0.251	0.272
	P/N	0.592	0.191	0.187	0.608	0.257	0.269