Table 1.

Primers used in this study to optimize one-step reverse transcription-recombinase polymerase amplification (RT-RPA) for detection of potato virus A (PVA)

Target virus	Primer set No	Primers	Polarity	Sequence 5ʹ–3ʹ	Bases	Target position on coat protein gene	Position on accession number in NCBI GeneBank (AJ131400.1)	GC (%)	Tm (°C)	Amplicon size (bp)
PVA	A	PVARPAF1	Sense	CTGAAGGTAAGAAGAAAGAAGGAGAAG	27	37–63	8572–8756	40.7	60.7	185a
		PVARPAR1	Antisense	GTAAGATAGCAAGTGATCTAGGTTAACGAC	30	192–221		40	61.3
	B	PVARPAF1	Sense	CTGAAGGTAAGAAGAAAGAAGGAGAAG	27	37–63	8572–8747	40.7	60.7	176
		PVARPAR2	Antisense	CAAGTGATCTAGGTTAACGACACTCTTAC	29	184–212		41.4	61.7
	C	PVARPAF2	Sense	GAAGGTAAGAAGAAAGAAGGAGAAGG	26	39–64	8574–8756	42.3	60.6	183
		PVARPAR1	Antisense	GTAAGATAGCAAGTGATCTAGGTTAACGAC	30	192–221		40	61.3

^aFor optimization and validation of one-step RT-RPA, the best primer pair was selected