Melecular epidemiology of PRRSV in Europe

Report
Impact of PRRSV diversity on diagnostic
activities
Tomasz Stadejek
Type 1 PRRSV (European) belongs to several
distinct subtypes
ORF5 identity down to
69.5% (Clustal V)
S u b typ e 3
21
29
30
40
18 9
22
7
10
11
32
ORF7 identity down to
75.4% (Clustal V)
87
19
66 (S ubtype 1)
49
28
96
24
27
34
23
39
97
S u b typ e 2
59
42
107,108 42
106
69
24
99
63
988
101
38
80
37
781
743
94
3
35
108 107
53
50
102
2
4
38
97
37,109
25
98
36
26 105
44
686
60
61
12
S u b typ e 1
43
56
54
71
89
90
61
16
48
47
93
71
89
88
Belarus
Lithuania
Western Europe, Asia, North America
85
843
75
343
35
91
90
92
77
56
66
55
53
7
74
64
83
94
14
84
82
69
62
78
76
36
67
68
79
81
92
Russia
86
S u b ty p e 2
72
93
44
87
533
816
65
83
33
57
933
45
95
18
27
45
103
17
22
21
46
43
1
20
9
55
100
S u b ty p e 3
19
8
52
327
41
109
106
25
26
40
54
28
104
13
15
6
10
S u b ty p e 1
5
31
41
95
15 13
14
16
50
70
63
12
5
73
51
80
58
59
Type 1, subtype 2: 378, 375
ORF7
polymorphism
Type 1, atypical?: 393
Type 1, subtype 4: 375
Type 1, subtype 1: 387, 390, 399
Type 1, subtype 3: 375, 387
Type 1, atypical?: 378
Type 2: 372, 375
+--------+--------30
40
+--------+--------subtype 1 Lelystad 1991 NL-387
AAAGAA---AAGTACAGCTCCGA
subtype 1_776 RO-390 variant
AAAAAACAAAAACACAGCTCCGA
subtype 1_28M_2009 SK-399 variant
GAAGAA---AAGTACAGCTCCAA
subtype 2 IL-1999 RU-378
AAATAA---AAATACAGCTCCAA
subtype 2 Ili 2_2009 RU-375 variant AAACAA---AAATACAGCTCCAA
subtype 3 Soz 2004 BY-375
AAAGAA---AAATACAGCTCCAA
subtype 3 Soz_2009_BY-387 variant
AAAGAA---AAGTACAGCTCCAA
subtype 4_Okt BY-375
AAAGAA---AAATGTAGCTCCAA
atyp Bor-54 BY-393
AAATAA---AAACACAGCTCTAA
atyp VL RU-378
ACAAATCAAAAACACAACTCCAA
+--------+--------130
140
+--------+--------GGACAGGC---CAAAAAGAAAAA
GGACAGGC---AAAAAGGAAAAA
GGACAGGC---CAGAAAGAAAAA
GGACGGGCTAATAAAAAGAGGAA
GGACGGGT---TAAAAAGAGGAA
GGACAGGA---AAAAAAAAAAAG
GGACAGGA---AAAAAGGAAAAG
GGACAGGC---CAAAAGGAAAAA
GGACAGGCTCACAAAAGGAAAAA
GGACAGGC---CAAAAAGAAAAG
+---------+---------+------360
370
380
+---------+---------+------TACATCCGCCAGTCAGGGTGCAAGTTAA
TGCATCCGCCAGTCAGGGTGCAAGTTAA
CACATCCGCGAGTCAGGGTGCAAGTCAATTTGGTAGTTAG
CGCATCTGCCAGTTAA
CGCATCCGCCAGTTAG
CACGCCCGTTAGTTAA
CACGCCCACTAGTCAAGGCGCAAGTTAA
TACATCCAATAGTTAG
CGCACCCAATGATTTGGGCGCAACTCAATAG
CGCGCCTGCTAGTTAA
LV 40
50
Lelystad 387sub1
QPRGGQA-KKKKP
Czechy_47 387sub1_070118
QPRGGQA-RKKRP
AS_34M 387sub1EE
QPRGGQA-KKKKP
Eigir LT 2006 378sub2
QPRGGQASKRKKP
Ili_6_100210 378sub2
QPRGGRVNKRKKP
Ili_9_100210 375sub2
QPRGGRV-KKRKP
Zad_1_041201 375sub3
QPRGGQA-KKKKP
Obu_1_050310 375sub3
QPRGGQA-KKKRP
Yuz_34_050310 375sub3
QPRGGQA-KKKRP
Soz_091120 387sub3STOPCOD
QPRRGQE-KRKRP
Soz 2A_85_85_060215 375sub3 QPRRGQG-IKKKP
Okt-46 375atyp
QPKGGQA-KRKKP
Bor_59 393atyp_091209
QPRGGQANKRKKP
34.9
Zve_091120 378sub2
Bel arus 3 378sub2 _050718
Svit_1_091120 378sub2
Sno_4_050211 378sub2
Ukraine 2_2_ 080929 378sub2
Ili_6_100210 378sub2
Ili_9_100210 375sub2DELETION
Syberia_2_070118 378sub2
Ei gir LT 2006 378sub2
Bel -42 378sub2
Soz 2_4_14_060215 378sub2
Bor_59 393atyp_091209
VL-2001_RU 378atyp
Amervac 387sub1
Porcil is 387sub1
Zalewski 56 387sub1
Konskowola 6 tyg 387sub1
Czechy_47 387sub1_070118
AS _34M 387sub1E E
Okt-46 375atyp
Soz 2A _48_70_060215 387sub3S TOPCOD
Soz_091120 387sub3S TOPCOD
Soz 2A _85_85_060215 375sub3
Mozy_100210 375sub3
Czys_1 375sub3_091209
ZAO TURIE C_1_ORF7 375sub3
Obu_1_050310 375sub3
Szaj _100210 375sub3
Zad_1_041201 375sub3
Yuz_34_050310 375sub3
Vos-29 375sub3
vr2332
30
25
20
15
10
Nucl eoti de S ubstituti on per 100 residues
5
0
2
1
3
Strain
Type 1
subtype
SDOW17
(EU+NA)
VO17 (NA)
WB4 (EU)
231/Sza
3
415/And
nd
6695/Bie
2
8380/Okt
4
Bor
atypical
+
+
+
+
+
+
+
+
+
-
Courtesy of Anette Botner
Belarusian subtype 3 PRRSV isolate
Vietnamese PHFD PRRSV isolate
NADIR experiment DTU Vet Lindholm,
Denmark
•
•
•
•
To assess virulence of east European variants of type 1 PRRSV
PAM isolation
Pigs free from 29 known pig pathogens, intranasally inoculated
4 groups
–
–
–
–
7 pigs inoculated with Danish isolate 18794 subtype 1
7 pigs inoculated with Russian isolate Ili subtype 2
7 pigs inoculated with Belarusian atypical isolate Bor
7 pigs sham inoculated
• Monitored for 24 dpi
9
VET LIndholm
9
Development of body temperature in pigs
infected with type 1 PRRSV strains
Danish(subtype 1)
18794
ILI(subtype 2)
Ili
Bor(atypical)
Bor
controls
Controls
oC
41.5
41.0
40.5
40.0
39.5
39.0
38.5
38.0
37.5
-4
-1
2
5
8
11
PID
14
17
20
23
26
Courtesy of Jens Nielsen
10
VET LIndholm
Can wild boars transmit
East European strains
westward?
+ Sid
+Au s
Lithuania
Upa*
* Sma
Prz *
* Ch e
+Vas
# Obu
x Okt
Sn o
+
Stankevicius et al., Barcelona 2011
Dzi
Kon
Bie
*
Bor
+
Gro Kwi
*
Sok
*
*
*
* Lek Poland
*
*
Rak
Krz
Gra
Po l
*
*
Zad
#
Vos
#
Belarus
*
+/# Bel
# Zap
Yuz
#
Soz
#
* Jed
* Nie
* Zbr
100 km
ESVV 2012, Sep 5, Madrid
11
NextGen PRRS (Tetracore)
Adiavet PRRS EU/NA (Adiagene)
TaqVet PRRS Triplex (LSI)
Toplak et al.: J Vir Meth
13
17454/5-1
100.0
2.5
% of positive samples
90.0
2
70.0
60.0
1.5
50.0
40.0
1
30.0
20.0
0.5
10.0
17454/5-3
0.0
0
4
100.0
90.0
80.0
70.0
60.0
50.0
40.0
30.0
20.0
10.0
0.0
6
2.5
8
10
12
14
16
18
age in weeks
2
1.5
1
0.5
0
4
6
8
10
12
14
age in weeks
% of positive samples
% of positive samples
80.0
16
1111
18
100.0
90.0
80.0
70.0
60.0
50.0
40.0
30.0
20.0
10.0
0.0
2.5
2
1.5
1
0.5
0
4
6
8
10
12
age in weeks
14
16
19
ESVV 2012, Sep 5, Madrid
15
Genotype 2 PRRSV in
Europe
Poland 98.7%
Spain 97.7%
Poland 98%
Germany 96%
Hungary 95.7%
Poland 96.2%
Poland 93.7%
Germany 91.2%
Hungary 87.7%
% identity to Ingelvac PRRS
MLV
Clustal W
Discriminatory ELISA in a herd co-infected with two
genotypes
17454/5-3
100.0
2.5
90.0
% of positive samples
80.0
2
70.0
60.0
1.5
PRRSV type 1
50.0
ELISA
40.0
1
PRRSV type 2
mean S/P
30.0
20.0
0.5
10.0
0.0
0
4
6
8
10
12
age in weeks
14
16
18
Jens Nielsen
Lars E. Larsen
Anette Botner
Lise Kvisgaard
Martin Oleksiewicz Gyula Balka
Torben Storgaard
Daniel Cadar
Arunas Stankevicius Gabi Schageman
Denis Potapchuk
Oliver Duran
Alexei Scherbakov Fred Leung
Ewelina Skrzypiec
Manreet Brar
Katarzyna Szymanek Jose Angulo
Michal Fabisiak
Mike Roof
Rafal Sapierzynski
Eric Nelson
Katarzyna Podgorska Rolf Rauh
Artur Jablonski
Marian Porowski
Iwona Stankiewicz
Thank you!
ESVV 2012, Sep 5, Madrid
19

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