Two clinical cases of renal syndrome caused by Dobrava/Saaremaa hantaviruses imported to the Netherlands from Poland and Belarus, 2012–2014

Figures & data

Table 1 Hantavirus diagnostics performed in convalescent sera of two patients in the Netherlands with HFRS after travelling to Belarus and Poland, 2012–2014

			Patient 1		Patient 2
			Day 10	Day 52	Day 3	Day 9
			After symptom onset		After symptom onset
ELISA ^a	PUUV	IgM	17.475	17.728	1.159	1.446
		IgG	11.96	12.104	0.295	2.392
IFA ^b	DOBV	IgM	4,000	1,000	100	2,000
		IgG	128,000	64,000	4,000	≥16,000
	SAAV	IgM	16,000	1,000	1,000	2,000
		IgG	4,000	64,000	4,000	≥16,000
	PUUV	IgM	<100	<100	<100	<100
		IgG	<100	<100	<100	100
FRNT ^c	DOBV			1:320		1:160
	SAAV			1:160		1:160
	PUUV			<40		<40
PCR ^d	DOBV		Negative		Positive
	PUUV		Negative		Negative
	SEOV		Negative		Negative

PUUV, Puumula virus; DOBV, Dobrava virus; SAAV, Saarema virus; SEOV, Seoul virus.

Bold figures indicate positivity. ^aEnzyme-linked immunosorbent assay (Progen), values <1.0 were considered negative (4)]. ^bIndirect immunofluorescence assay (Euroimmuun), titers <100 were considered negative (5). ^cVirus neutralization test, titers <40 were considered negative (1)]. ^dIn-house real-time reverse transcriptase-polymerase chain reaction.

Table 2 Primer and probes used for internally controlled DOBV real-time RT-PCR

	Name	Sequence 5′-3′	Conc (pmol/µl)	References
Dobrova	Dobrava -rev-TM	AGACATTCAGGAAGCAAATYAATGA	30	In-house
	Dobrava-fwd-short	GGTGGTTTAGGAYGTCACCTTAAGTG	45
	Dobrava-probe-new	FAM-ACAACAACTAYCTACC^aCAAAACAACAACACTACCTCA	10
PDV	PDV fwd	CGGGTGCCTTTTACAAGAAC	30	(3)
	PDV rev	TTCTTTCCTCAACCTCGTCC	7.5
	PDV probe	CY5-ATGCAAGGGCCAATT-MGB	10

^aBHQ1, internally coupled quencher. PDV, phocine distemper virus, used as an internal process control.

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