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Volume 20, Number 12—December 2014
Dispatch

Mycobacterium Species Related to M. leprae and M. lepromatosis from Cows with Bovine Nodular Thelitis

Didier PinComments to Author , Véronique Guérin-Faublée, Virginie Garreau, Franck Breysse, Oana Dumitrescu, Jean-Pierre Flandrois, and Gerard Lina
Author affiliations: VetAgro Sup Campus Vétérinaire de Lyon, Marcy l’Étoile, France (D. Pin, V. Guérin-Faublée); Clinique Vétérinaire, Saint Bénigne, France (V. Garreau); Centre Hospitalier Lyon Sud, Pierre Bénite, France (F. Breysse); Université Lyon 1, Lyon, France (O. Dumitrescu, J.-P. Flandrois, G. Lina)

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Table

Primers used for PCR detection of Mycobacterium species, Jura, France

Gene*
 Sequence (5′→3′)†
16S rRNA gene (6) F-TCAAAKgAATTgACgggggC
R-ggTTACCTTgTTACgACTT
hsp65 (5) F-ACCAACgATggTgTgTCCAT
R-CTTgTCgAACCgCATACCCT
rpoB (designed for this study) 2F-TCAACgggACCgAgCgTgTC
2R-gTgTTgTCCTTCTCCAgCgT
3F-TCAACgggACCgAgCgTgTC
4R-gTCTCgATCgggCACATC
9F-gTgggCACCggCATggAgTT
9R-ATgTTCATCCgTCgCggC
8F-ATGAAgCTgCACCACTTggT
8R-gCCGATTCgTTgCgggACA
sodA (5) F-AgCTTCACCACAgCAAgCACCA
R-TCggCCAgTTCACgACgTTCCA
tuf (5) F-CACgCCgACTACATCAAgAA
R-gAACTgCggACggTAgTTgT
tmRNA (5) F-ggggCTgAAACggTTTCgAC
R-TggAgCTgCCgggAATCgAAC
*hsp65, heat shock protein 65; rpoB, β-subunit of RNA polymerase; sodA, superoxide dismutase A; tuf, elongation factor Tuf; tmRNA, transfer–messenger RNA. 
†F, forward; R, reverse. DNA was amplified by using a LightCycler 2.0 instrument (Roche Molecular Diagnostics, Indianapolis, IN, USA) and a ready-to-use hot start reaction mixture (TAKARA, Tokyo, Japan). For all PCRs, parameters were denaturation 95°C for 30 s; and amplification (45 cycles) at 95°C for 15 s, 65°C for 20 s, and 72°C for 40 s (slope, 2°C/s); melting curve, 0 s at 95°C, 15 s at 75°C, and 0 s at 98°C (slope, 0.05°C/s).

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