Table 1.

 Details of reverse transcriptase‐polymerase chain reactions (RT‐PCRs) for the detection of bovine torovirus‐, bovine coronavirus‐ and rotavirus‐specific nucleic acids

	Bovine torovirus	Bovine coronavirus	Rotavirus
Amplification product	741 bp	407 bp	433 bp
Upstream primer	5′‐GTG TTA AGT TTG TGC AAA AAT G‐3′	5′‐GCC GAT CAG TCC GAC CAA TC‐3′	5′‐AAG TAG CTG GAT TTG ATT ATT C‐3′
Downstream primer	5′‐TGC ATG AAC TCT ATA TGG TGT‐3′	5′‐AGA ATG TCA GCC GGG GTA T‐3′	5′‐GAC TCA CAA ACT GCA GAT TCA A‐3′
Volume template	2.5 μl	1.6 μl	1.5 μl
Volume RT‐PCR mixture	22.5 μl	18.4 μl	13.5 μl
Reverse transcription	50°C, 30 min	50°C, 30 min	50°C, 30 min
Initial denaturation	95°C, 15 min	95°C, 15 min	95°C, 15 min
Number of cycles	35	35	40
Temperature profile	95°C, 1 min	94°C, 30 s	94°C, 1 min
	55°C, 1.5 min	58°C, 1 min	55°C, 1 min
	72°C, 1.5 min	72°C, 2 min	72°C, 1 min
Final extension	72°C, 10 min	72°C, 10 min	72°C, 7 min