Evaluation of a real time pcr to simultaneously detect and differentiate virulent and non-virulent Newcastle disease virus



Yüklə 421,27 Kb.
səhifə3/4
tarix05.05.2017
ölçüsü421,27 Kb.
1   2   3   4

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


Yüklə 421,27 Kb.

Dostları ilə paylaş:
1   2   3   4




Verilənlər bazası müəlliflik hüququ ilə müdafiə olunur ©azkurs.org 2020
rəhbərliyinə müraciət

    Ana səhifə