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Journal of Clinical Microbiology, October 2003, p. 4907-4908, Vol. 41, No. 10
0095-1137/03/$08.00+0 DOI: 10.1128/JCM.41.10.4907-4908.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
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Of 864 patients who attended a referral clinic of the National TB Program in Yangon in July 2000, 202 were diagnosed with pulmonary TB based on medical history, clinical signs, two smear-positive sputum samples, and a chest X-ray. Of these, approximately half were new cases (no previous TB treatment) and isolates from 51 cases were available for the present study. All isolates belonged to the M. tuberculosis complex (AccuProbe Mycobacterium tuberculosis complex test; Gen-Probe, San Diego, Calif.). Standard biochemical assays (thiophen-2-carboxylic acid hydrazide [TCH] susceptibility test, niacin accumulation test, nitrate reduction test, and pyrazinamidase activity) were performed for phenotyping (4). Spoligotyping was performed with a commercial kit as recommended by the manufacturer (Isogen Bioscience BV, Maarssen, The Netherlands). IS6110 restriction fragment length polymorphism (RFLP) analyses were performed as previously described (8). The phenotype variants, spoligotype families, and IS6110 RFLP dendrogram of the isolates are shown in Fig. 1. The majority of the isolates belonged phenotypically to the classical and Asian human M. tuberculosis variants as previously defined by Collins et al. (2). The spoligotype families were defined in accordance with Sebban et al. (6), but three isolates did not correspond to any of the defined families. When the RFLP patterns of the isolates with >65% Dice coefficient similarity were examined, a criterion used previously to discern IS6110 groups (9), nine groups (A to I) could be discriminated.
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FIG. 1. Dendrogram of 51 isolates produced following Dice and unweighted pair group method with arithmetic average analysis of IS6110 RFLP and distribution of the isolates according to spoligotype family and phenotype variants. Spoligotype families are according to Sebban et al. (6). UR, previously unreported spoligotype families. a, shared type 292; b, variant 1, Asian human M. tuberculosis variant (susceptible to TCH, positive niacin accumulation, nitrate reduction, and pyrazinamidase activity tests); variant 7, classical human M. tuberculosis variant (resistant to TCH, positive niacin accumulation, nitrate reduction, and pyrazinamidase activity tests); variants 2 to 6, variants between classical and Asian human M. tuberculosis strains.
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Fifteen of the 18 Beijing family isolates belonged to RFLP group A, whereas the EAI family isolates were distributed among RFLP groups A, B, C, E, H, and I. Moreover, the Beijing family isolates belonged predominately to the classical human M. tuberculosis variant while the EAI family isolates belonged predominately to the Asian human M. tuberculosis variant (P = 0.02, chi-square test). These differences suggest that the two spoligotype families may have evolved differently in Myanmar and that the majority of the Beijing family strains seem to belong to a distinct genetic lineage. The high heterogeneity of the RFLP patterns of the isolates indicates, however, that several strains contribute to the TB endemicity in Yangon.
We thank Christophe Sola, Nalin Rastogi, and coworkers (Unité de la Tuberculose et des Mycobactéries, Institut Pasteur de Guadeloupe, Guadeloupe) for sharing their extensive spoligotype database with us.
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Sabai Phyu*
Department of Microbiology and Immunology The Gade Institute University of Bergen Bergen, Norway
Roland Jureen
Ti Ti
Ulf R. Dahle
Harleen M. S. Grewal
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| * Phone: 47-55975542Fax: 47-55974979E-mail: sabai.phyu{at}cih.uib.no |
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