Table 2 continued……..
-
Sample Number
|
Species
|
Virulent/Avirulent APMV-1 qPCR
|
APMV-1 Multiplex qPCR
|
Ct# ARP2 Probe
|
Ct VRP2 Primer
|
Ct APMV Wise
|
Ct APMV Kim
|
46
|
Chicken
|
28.29*
|
>40
|
28.13
|
>40
|
47
|
Chicken
|
>40
|
>40
|
>40
|
>40
|
48
|
Pigeon
|
>40
|
>40
|
>40
|
>40
|
49
|
Chicken
|
>40
|
>40
|
>40
|
>40
|
50
|
Chicken
|
>40
|
>40
|
30.3
|
>40
|
51
|
Chicken
|
28.88
|
>40
|
26.39
|
>40
|
52
|
Chicken
|
>40
|
>40
|
>40
|
>40
|
53
|
Chicken
|
>40
|
>40
|
>40
|
>40
|
54
|
Chicken
|
>40
|
>40
|
>40
|
>40
|
55
|
Chicken
|
>40
|
>40
|
29.4
|
>40
|
56
|
Chicken
|
>40
|
>40
|
>40
|
>40
|
57
|
Chicken
|
>40
|
>40
|
>40
|
>40
|
58
|
Chicken
|
>40
|
>40
|
>40
|
>40
|
59
|
Chicken
|
33.68
|
>40
|
27.73
|
>40
|
60
|
Chicken
|
>40
|
>40
|
>40
|
>40
|
61
|
Chicken
|
>40
|
>40
|
>40
|
>40
|
62
|
Chicken
|
>40
|
>40
|
>40
|
>40
|
* Confirmed as being avirulent based upon sequence analysis of the fusion gene
# Ct refers to the crossing threshold value in Real time PCR. Typically a value >40 is negative, and Ct values < 36-40 are positive (this value can vary depending on the test).
Table 3: Virulent/avirulent APMV-1 qPCR results for a range of wild bird samples that tested positive in the Wise/Kim qPCR test used to pre-screen the samples for the presence of APMV-1.
Sample Number#
|
Species
|
APMV-1 Multiplex qPCR
|
Virulent/Avirulent APMV-1 qPCR
|
APMV Kim qPCR
Ct
|
APMV Wise qPCR
Ct
|
Ct ARP2 Probe
|
Ct VRP2 Primer
|
578
|
Grey Teal
|
>40
|
32.18
|
>40
|
>40
|
589
|
Wood Duck
|
>40
|
37.89
|
>40
|
>40
|
595
|
Wood Duck
|
23.52
|
>40
|
>40
|
>40
|
596
|
Grey Teal
|
>40
|
37.37
|
>40
|
>40
|
642
|
Chestnut Teal
|
30.45
|
33.80
|
>40
|
>40
|
657
|
Chestnut Teal
|
30.91
|
38.59
|
>40
|
>40
|
800
|
Pacific black duck
|
28.12
|
>40
|
>40
|
>40
|
805
|
Pacific black duck
|
32.56
|
>40
|
>40
|
>40
|
816
|
Pacific black duck
|
35.56
|
>40
|
>40
|
>40
|
848
|
Duck
|
>40
|
30.97
|
>40
|
>40
|
856
|
Duck
|
33.66
|
31.87
|
>40
|
>40
|
858
|
Duck
|
>40
|
29.31
|
>40
|
>40
|
859
|
Duck
|
>40
|
33.10
|
>40
|
>40
|
866
|
Duck
|
33.10
|
29.74
|
>40
|
>40
|
873
|
Duck
|
>40
|
32.89
|
>40
|
>40
|
875
|
Duck
|
25.07
|
31.58
|
>40
|
>40
|
896
|
Duck
|
>40
|
29.36
|
>40
|
>40
|
897
|
Duck
|
34.68
|
30.30
|
>40
|
>40
|
898
|
Duck
|
>40
|
35.10
|
>40
|
>40
|
902
|
Duck
|
>40
|
31.02
|
>40
|
>40
|
909
|
Duck
|
>40
|
31.02
|
>40
|
>40
|
912
|
Duck
|
>40
|
33.10
|
>40
|
>40
|
921
|
Duck
|
32.89
|
28.60
|
>40
|
>40
|
923
|
Duck
|
>40
|
31.57
|
>40
|
>40
|
930
|
Duck
|
34.41
|
>40
|
>40
|
>40
|
931
|
Duck
|
>40
|
30.80
|
>40
|
>40
|
975
|
Pink ear
|
34.12
|
>40
|
>40
|
>40
|
986
|
Grey Teal
|
27.92
|
>40
|
>40
|
>40
|
995
|
Pink ear
|
>40
|
36.51
|
>40
|
>40
|
1001
|
Grey Teal
|
30.83
|
>40
|
>40
|
>40
|
1008
|
Grey Teal
|
23.02
|
>40
|
>40
|
>40
|
429 samples
|
Mixed duck
|
>40
|
>40
|
NT
|
NT
|
# Sample numbers are the same as used in the Avian influenza project and have not been renumbered for this project to avoid confusion.
NT Not tested
Red Also Positive in AIV PCR test
Table 4: Virulent/avirulent APMV-1 qPCR and nested Virulent/avirulent APMV-1 qPCR results for a set of control samples positive in the Wise/Kim qPCR test used to pre-screen the samples for the presence of APMV-1.
Samples
|
Class
|
Vir/Avir
|
Virulent/Avirulent APMV-1 qPCR
|
Nested Virulent/Avirulent APMV-1 qPCR
|
APMV-1 Multiplex qPCR
|
ARP
|
VRP
|
ARP
|
VRP
|
APMV Kim qPCR
|
APMV Wise qPCR
|
Strong Positive control
|
II
|
A
|
28.50
|
>40
|
5.19
|
>40
|
>40
|
24.67
|
Weak Positive control
|
II
|
A
|
38.56
|
>40
|
17.53
|
>40
|
>40
|
30.74
|
NDV NQC replicate 1
|
II
|
V
|
>40
|
>40
|
>40
|
16.47
|
>40
|
32.55
|
NDV NQC replicate 2
|
II
|
V
|
>40
|
>40
|
>40
|
15.70
|
>40
|
32.39
|
NDV NQC replicate 3
|
II
|
V
|
>40
|
>40
|
>40
|
35.86
|
>40
|
32.42
|
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