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Journal of Clinical Microbiology, January 1998, p. 239-242, Vol. 36, No. 1
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Allele-Specific PCR Method Based on pncA
and oxyR Sequences for Distinguishing Mycobacterium
bovis from Mycobacterium tuberculosis:
Intraspecific M. bovis pncA Sequence
Polymorphism
Luz Elena Espinosa
de los
Monteros,1,
Juan Carlos
Galán,1
Montserrat
Gutiérrez,2
Sofía
Samper,3
Juan F.
García
Marín,2
Carlos
Martín,3
Lucas
Domínguez,4
Luis
de Rafael,1
Fernando
Baquero,1
Enrique
Gómez-Mampaso,1 and
Jesús
Blázquez1,*
Servicio de Microbiología, Hospital
Ramón y Cajal, Instituto Nacional de la
Salud,1 and
Departamento de
Patología Animal I (Sanidad Animal), Facultad de Veterinaria,
Universidad Complutense,4 Madrid,
Anatomía Patológica, Departamento de
Patología Animal, Facultad de Veterinaria, Universidad de
León, León,2
and
Departamento de Microbiología y Medicina
Preventiva, Facultad de Medicina, Universidad de Zaragoza,
Zaragoza,3 Spain
Received 12 May 1997/Returned for modification 2 September
1997/Accepted 21 October 1997
An allele-specific amplification method based on two genetic
polymorphisms to differentiate Mycobacterium tuberculosis
from Mycobacterium bovis was tested. Based on the
differences found at position 169 in the pncA genes from
M. tuberculosis and M. bovis, a PCR system
which was able to differentiate most of the 237 M. tuberculosis complex isolates tested in one of the two species
was developed. All 121 M. tuberculosis strains showed the
expected base (cytosine) at position 169. Most of the M. bovis isolates had a guanine at the cited position. Nevertheless,
18 of the 116 M. bovis isolates, all of them goat isolates,
showed the pncA polymorphism specific to M. tuberculosis. These results suggest that goat M. bovis may be the nicotinamidase-missing link at the origin of the
M. tuberculosis species. Based on the polymorphism found at
position 285 in the oxyR gene, the same system was used to
differentiate M. tuberculosis from M. bovis. In
this case, DNAs from all 121 M. tuberculosis isolates had
the expected base (guanine) at this position. In addition, all 116 M. bovis isolates, including those from goats, showed the
identical polymorphism (adenine). The oxyR allele-specific
amplification method can differentiate M. bovis from
M. tuberculosis, is rapid (results can be obtained in less
than 3 h), and is easy to perform.
*
Corresponding author. Mailing address: Servicio de
Microbiología, Hospital Ramón y Cajal, Ctra. Colmenar km
9.100, 28034 Madrid, Spain. Phone: 34-1-336 83 30. Fax: 34-1-336 88 09. E-mail: jesus.blazquez{at}hrc.es.
Present address: Servicio de Microbiología, Hospital
Infantil de México Federico Gómez, México 06720 D.F., México.
Journal of Clinical Microbiology, January 1998, p. 239-242, Vol. 36, No. 1
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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