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

Table 5

Effect of the matrix on qRT-PCR performance.

(a)

RABV (TCID50)	CSF	Saliva	Urine
1.00𝐸+05	19.65	18.80	18.53
1.00𝐸+04	24.50	23.39	23.13
1.00𝐸+03	28.12	27.17	26.85
1.00𝐸+02	33.15	32.75	30.56
1.00𝐸+01	40.53	ND	39.67
1.00𝐸+00	43.54	ND	39.78
1.00𝐸−01	ND	ND	ND
1.00𝐸−02	ND	ND	ND

(b)

EBLV-1 (TCID50)	CSF	Saliva	Urine
1.00𝐸+04	14.56	15.31	14.50
1.00𝐸+03	18.11	18.34	18.84
1.00𝐸+02	22.72	23.79	22.17
1.00𝐸+01	27.74	28.90	28.06
1.00𝐸+00	33.10	28.17	28.67
1.00𝐸−01	ND	ND	ND
1.00𝐸−02	ND	ND	ND

Saliva, CSF, and urine samples were spiked with decreasing doses of RABV or EBLV-1. Three independent runs with each time two repeats were performed per virus dose (10-fold serial dilution). The limit of detection for RABV was 10⁰ TCID₅₀ in urine, 10¹ TCID₅₀ in CSF, and 10² TCID⁵⁰ in saliva. For EBLV-1, the limit of detection was 10⁰ TCID₅₀ in CSF, saliva, and urine. ND: signal not detected.