TABLE 3.

qPCR assays to detect Pvrbps

Target gene	Primers and probea	Positionb (base)	R2	Effc (%)	Detection limit (copies/reaction)
Pvama1	CCTACTTCAGGAAGAAGGCTAACAAT	1514	0.999	107.5	<5
	GGTGGTGGGCTTCCCGTA	1590
	HEX-CCCTCTGCCTGGTCCATCTTGTCATACTT-BHQ1	1571
Pvdbp	TTAAAGACAAGACTTACTTTGCCGC	3146	0.999	105.8	<5
	TTTGATCATCTTCCTTGAAGCAATTAA	3330
	HEX-AACAGCAGTATCAGCAACGCTCCCG-BHQ1	3302
Pvrbp1a	GCATACAGAGAACACCCAGGATG	8417	0.999	98.7	<5
	GCTCCTCTTGGTCATCTTTCTTATGT	8598
	HEX-CCTATCAGGATACGTCAAATTCCAGCGATG-BHQ1	8443
Pvrbp1b	AGCAGGCTAAGACAGTTGCGAA	7408	0.999	100.0	<5
	AATGTCCTTCTTCTTCAGGTCAATTC	7529
	HEX-TCCAGCATGGCTTCATTCTCCCTTATATG-BHQ1	7501
Pvrbp2a	GGCAAGGCAAATTGTTCAGC	7321	0.999	102.5	<5
	ACGGACATTTCCATTTTCATTTCT	7442
	HEX-CCTCTTGAATGGTTTCCACTTCCATAATGG-BHQ1	7341
Pvrbp2b	CACAACATGAGGATCAGTCACAAC	8468	0.997	98.9	<5
	ATACCTGAAAAGACAGATAATCCCAAA	8561
	HEX-AATTCCTYCTGCAAGGCGAACTCGAC-BHQ1	8532
Pvrbp2c	CGCATGACGATACGCATGAC	8414	0.999	102.8	<5
	CCTGCTAAACGAGTCTTTCCAATT	8495
	HEX-TTTGCCGTTGAATCCCTTCCASTTTGT-BHQ1	8465
Pvrbp2p1	TTGTTTCTACTTGTGAAGAAGCCAAA	1622	0.998	100.0	<5
	CATCATAACTYTGTTTAACTGATGCAATT	1747
	HEX-TAGAATCCTTGCGTACYTCGAGCACCG-BHQ1	1655
Pvrbp2d	CAGAAGAGAAACTTTAGAAAGTGACCT	7402	0.999	100.8	<5
	CTTCACTTAATGTATTCGCATTTAATT	7487
	HEX-TTTCACATTTGTGTCCTCTWCATGTCCTTT-BHQ1	7459
Pvrbp2e	CGACTGCTACAAACATATTAGATAACACA	4336	0.999	101.1	<5
	TCTGAGCTACCGCTTCTTCTAATAATT	4446
	HEX-TGATTTCTCAAGTTCCTTTGCCGCCTC-BHQ1	4415
Pvrbp3	CTGGTGACAACCATTCTGATGAC	8413	0.999	97.3	<5
	AAATCGGCTTCTTGTGATTCATG	8599
	HEX-TTAATGATTGCGAATCCAGCACTTGAGC-BHQ1	8566

^aThe sequences of primers and probes are shown from 5′ to 3′.

^bThe position of the 5′ end of each oligonucleotide is relative to the start codon in the Sal-1 gene sequences, except for Pvrbp2e. Primer and probe locations of Pvrbp2e are based on the VTTY57 isolate.

^cEff, PCR amplification efficiency.