Direito

Molecular and Cellular Probes 20 (2006) 182–190 www.elsevier.com/locate/ymcpr

Study of the gyrB gene polymorphism as a tool to differentiate among Mycobacterium tuberculosis complex subspecies further underlines the older evolutionary age of ‘Mycobacterium canettii’
Khye Seng Goh a, Michel Fabre b, Richard C. Huard c,1, Solveig Schmid a, Christophe Sola a, Nalin Rastogi a,*
´ ´ ` ` Unite de la Tuberculose et des Mycobacteries, Institut Pasteur de Guadeloupe, Pointe-a-Pitre, Guadeloupe, Morne Joliviere, BP484, F97183 Abymes, Cedex, Guadeloupe b ´ Laboratoire de Mycobacteriologie, HIA Percy, 92140 Clamart, France c Division of International Medicine and Infectious Diseases, Department of Medicine, Joan and Sanford I. Weill Medical College, Cornell University, New York, NY, USA Received 13 September 2005; accepted for publication 21 November 2005 Available online 6 March 2006 a Abstract The present investigation evaluated the PCR-restriction fragment length polymorphism (RFLP) analysis of hsp65 and gyrB targets for differentiation of the species within the Mycobacterium tuberculosis complex (MTC) both by including new restriction enzymes and previously unstudied species. The hsp65 restriction analysis using HhaI resulted in a characteristic ‘Mycobacterium canettii’ pattern. A study of the gyrB gene polymorphism using TaqIa and HinfI allowed the initial division of MTC into two major groups, one consisting of M. tuberculosis and ‘M. canettii’ as opposed to another single group with other species. Three different patterns were observed with RsaI, the first characteristic of Mycobacterium microti, the second with Mycobacterium bovis, M. bovis BCG and Mycobacterium caprae (M. caprae was easily separated from M. bovis, and M. bovis BCG by SacII digestion), and the third with M. tuberculosis, ‘M. canettii’, Mycobacterium africanum, Mycobacterium pinnipedii, and the dassie bacillus. Although further discrimination within the last group was not obtained using