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Journal of Clinical Microbiology, May 2002, p. 1798-1804, Vol. 40, No. 5
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.5.1798-1804.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Molecular Characterization of Pigmented and Nonpigmented Isolates of Mycobacterium avium subsp. paratuberculosis

Karen Stevenson,1* Valerie M. Hughes,1 Lucía de Juan,2 Neil F. Inglis,1 Frank Wright,3 and J. Michael Sharp1

Moredun Research Institute, Penicuik EH26 0PZ,1 Biomathematics and Statistics Scotland, Scottish Crop Research Institute, Dundee DD2 5DA, Scotland, United Kingdom,3 Departamento de Patología Animal I (Sanidad Animal), Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain2

Received 10 December 2001/ Returned for modification 3 February 2002/ Accepted 16 February 2002

Five pigmented isolates of Mycobacterium avium subsp. paratuberculosis were examined by pulsed-field gel electrophoresis (PFGE), IS900 restriction fragment length polymorphism (IS900-RFLP), and IS1311 polymorphism analysis using PCR. All of the pigmented isolates exhibited one of three distinct PFGE profiles with SnaBI, designated 9, 10, and 11, and with SpeI, designated 7, 8, and 9, which generated three multiplex profiles designated [9-7], [10-8], and [11-9]. All of the pigmented isolates had the same IS900-RFLP BstEII and PvuII profiles. The IS900-RFLP BstEII profile was new, but the IS900-RFLP PvuII profile corresponded to PvuII type 6 of a sheep strain described by Cousins and colleagues (D. V. Cousins, S. N. Williams, A. Hope, and G. J. Eamens, Aust. Vet. J. 78:184-190, 2000). IS1311-PCR analysis typed all of the pigmented isolates as sheep (S) strains. The genetic relationship between pigmented and nonpigmented isolates was investigated by using multiplex PFGE data from the analysis of both the 5 pigmented isolates and 88 nonpigmented isolates of M. avium subsp. paratuberculosis from a variety of host species and geographic locations. It was possible to classify the isolates into two distinct types designated type I, comprising the pigmented isolates, and type II, comprising the nonpigmented isolates, which exhibit a very broad host range.

* Corresponding author. Mailing address: Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik EH26 0PZ, Scotland. Phone: 44 131 445 5111. Fax: 44 131 445 6111. E-mail: stevk{at}mri.sari.ac.uk.

Journal of Clinical Microbiology, May 2002, p. 1798-1804, Vol. 40, No. 5
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.5.1798-1804.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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