TABLE 4.

Typing of aquabirnaviruses by the two-step restriction analysis assay^a

First step					Second step
Restriction endonuclease employed	Restriction fragment profile no.	No. of sitesb	Fragment size(s) (bp)	Reference strain (serotype)c	Restriction endonuclease employed	Fragment profile no.	No. of sitesb	Fragment size(s) (bp)	Reference strain (serotype)c
PvuII	1	0	1,179	WB (A1)	EcoRI	1	0	1,179	He (A4)
				He (A4)		2	1	979, 200	WB (A1)
	2	1	745	Te-2 (A5)	BstEII	1	0	1,179	Te-2 (A5)
			434	C1 (A6)		2	1	787, 392	C1 (A6)
	3	1	622 557	Ja (A9)	NNd				Ja (A9)
	4	2	557	Ab (A3)	BstII	1	0	1,179	C2 (A7)
			434	C2 (A7)		2	1	787, 392	Ab (A3)
			188	C3 (A8)		3	1	743, 436	C3 (A8)
	5	3	434	Sp (A2)	MboI	6	5	434, Snfe	TV-1 (B1)
			433	TV-1 (B1)		7	6	321,297, Smff	Sp (A2)
			188
			124

^aIn step 1 the first endonuclease reaction was applied to the initial 1,179-bp cDNA fragment. In step 2 the second endonuclease reaction was applied to the initial 1,179-bp cDNA fragment.

^bNumber of specific restriction sites in the initial cDNA fragment.

^cReference strain (corresponding serotype) showing the corresponding specific RFLP.

^dNN, not needed.

^eSnf, five fragments smaller than 240 bp, with 745 bp added.

^fSnf, five small fragments around 100 to 150 bp long, representing a total of 561 bp.