Table 1 Primer sequences and assay parameters of quantitative polymerase chain reactions for selected gene transcripts of warthogs.

From: Cytokine gene expression assay as a diagnostic tool for detection of Mycobacterium bovis infection in warthogs (Phacochoerus africanus)

Gene

Primer sequence (5′-3′)

Reverse

Tannealing

Pig identity

E%

AF

R2

IVA (%)

DM peak (°C)

Forward

B2M

CTCACTGTCTGGCCTGGATG

GGCGGATGGAACCCAGATAC

60 °C

F: 100%; R: 100%

103

2.03

0.99

0.5

82.5

H3F3A

AAACAGATCTGCGCTTCCAG

ACGTTTGGCATGGATAGCAC

60 °C

F: 100%; R: 100%

105

2.05

0.99

0.4

80.5

LDHA

TGCAACATGGCAGCCTTTTC

ACAACCAGCCTAGAGTTTGC

62 °C

F: 100%; R: 100%

111

2.11

0.99

0.5

79.5

PPIA

TGAGTGGTTGGATGGCAAAC

TGGTCTTGCCATTCCTGGAC

60 °C

F: 100%; R: 100%

114

2.14

0.99

0.7

77.5

YWHAZ

TTCTGAACTCCCCAGAGAAAGC

GCGTGCTGTCTTTGTATGACTC

62 °C

F: 100%; R: 100%

109

2.09

0.99

0.3

77.5

CXCL9

TCATCTTCCTGACTCTGCTTGG

TGGATCATCCTTTGGCTGGTG

62 °C

F: 95%; R: 100%

97

1.97

0.99

0.8

77.0

CXCL10

CCCACATGTTGAGATCATTGCC

TCTCTCTGTGTTCGAGGAGATC

60 °C

F: 77%; R: 100%

108

2.08

0.99

0.5

76.5

CXCL11

AAAGCGGGAAGGTGTCTTTG

GGCATCTTCGTCCTTTATGTGC

62 °C

F: 100%; R: 100%

105

2.05

0.99

0.7

78.0

IFNG

AGGCCATTCAAAGGAGCATG

AGTTCACTGATGGCTTTGCG

60 °C

F: 100%; R: 100%

115

2.15

0.98

1.0

78.5

TNFA

GGCCCAAGGACTCAGATCATC

ATACCCACTCTGCCATTGGAG

62 °C

F: 100%; R: 100%

101

2.01

0.98

1.0

81.5

  1. E%: Amplification efficiency; AF: Amplification factor; IVA: Intra-assay variability; DM: Derivative melt curve peak temperature; F: Forward primer identity; R: Reverse primer identity.