A Two-Step Lyssavirus Real-Time Polymerase Chain Reaction Using Degenerate Primers with Superior Sensitivity to the Fluorescent Antigen Test

Table 4

Comparison of the sensitivity between the two-step lyssavirus qRT-PCR and the one-step qRT-PCR published by Hayman et al. in 2011 [33].

Dilutions	RABV strain		EBLV-1 strain
	Two-step developed assay	One-step published assay	Two-step developed assay	One-step published assay
	Cq value	Cq value	Cq value	Cq value
1.00𝐸+00	9.52	17.41	6.77	19.51
1.00𝐸−02	12.58	23.80	11.40	26.92
1.00𝐸−03	17.59	27.31	17.07	30.70
1.00𝐸−04	21.97	30.95	21.37	33.70
1.00𝐸−05	26.16	34.29	24.98	37.36
1.00𝐸−06	30.94	38.10	28.35	ND
1.00𝐸−07	34.30	ND	ND	ND

RNA was extracted from infected cell culture supernatants, serially diluted and used to generate cDNA. The cDNA samples were tested by both qRT-PCR protocols. ND: no signal detected.