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Table 1 The primers and probes of the targeted genes in this study

From: Real-time PCR using atpE, conventional PCR targeting different regions of difference, and flow cytometry for confirmation of Mycobacterium bovis in buffaloes and cattle from the Delta area of Egypt

Type of PCR

Target gene

Primers and probe sequences

(5′-3′)

Amplicon size (bp)

Annealing temperature (°C)

Melting temperature (°C)

Accession number

Reference

Real-time

atpE

F 5′-CGGGCCGGATCGGGA-3′

R 5′-CGAAGACGAACAGCCAT-3′

P FAM 5′-ACGTGATGAAGAACGGGT AA-3′

182

60

63.7

57.1

58.9

CP023630.1

[22]

Conventionala

RD1

5′-AAGCGGTTGCCGCCGACCGACC-3′

5′-CTGGCTATATTCCTGGGCCCGG-3′

5′-GAGGCGATCTGGCGGTTTGGGG-3′

146

62

60.3

63.4

66

CP009186.1

[32]

RD4

5′-ATGTGCGAGCTGAGCGATG-3′

5′-TGTACTATGCTGACCCATGCG-3′

5′-AAAGGAGCACCATCGTCCAC-3′

172

67.5

66.7

67.1

CP009186.1

RD9

5′-CAAGTTGCCGTTTCGAGCC-3′

5′-CAATGTTTGTTGCGCTGC-3′

5′-GCTACCCTCGACCAAGTGTT-3′

235

66.6

63.9

66.5

CP009186.1

RD12

5′-GGGAGCCCAGCATTTACCTC-3′

5′-GTGTTGCGGGAATTACTCGG-3′

5′-AGCAGGAGCGGTTGGATATTC-3′

369

 

67

65.7

66.9

CP009186.1

  1. aRD-based detection of M. bovis as follows; RD1 present (146 bp), RD4 absent (268 bp), RD9 absent (108 bp), and RD12 absent (306 bp)