This Article Full Text Full Text (PDF) Supplemental material Other Versions of this Article: JCM.02231-07v1 46/4/1259    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Gibson, A. L. Articles by Ho, J. L. Search for Related Content PubMed PubMed Citation Articles by Gibson, A. L. Articles by Ho, J. L.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, April 2008, p. 1259-1267, Vol. 46, No. 4
0095-1137/08/$08.00+0     doi:10.1128/JCM.02231-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Application of Sensitive and Specific Molecular Methods To Uncover Global Dissemination of the Major RD^Rio Sublineage of the Latin American-Mediterranean Mycobacterium tuberculosis Spoligotype Family ^,

Andrea L. Gibson,^1, Richard C. Huard,^1,2, Nicolaas C. Gey van Pittius,³ Luiz Claudio Oliveira Lazzarini,^1,4 Jeffrey Driscoll,⁵ Natalia Kurepina,⁶ Thierry Zozio,⁷ Christophe Sola,^7,8 Silvana Miranda Spindola,⁹ Afrânio L. Kritski,⁴ Daniel Fitzgerald,^1,10 Kristin Kremer,¹¹ Helmi Mardassi,¹² Poonam Chitale,¹ Jessica Brinkworth,^1,13 Dario Garcia de Viedma,^14,15 Brigitte Gicquel,⁸ Jean W. Pape,¹⁰ Dick van Soolingen,¹¹ Barry N. Kreiswirth,⁶ Robin M. Warren,³ Paul D. van Helden,³ Nalin Rastogi,⁷ Philip N. Suffys,¹⁶ Jose Lapa e Silva,⁴ and John L. Ho^1*

Department of Medicine, Division of International Medicine and Infectious Diseases, Weill Medical College of Cornell University, New York, New York,¹ Clinical Microbiology Service and the Department of Pathology, Columbia University Medical Center, New York-Presbyterian Hospital, New York, New York,² DST/NRF Centre of Excellence in Biomedical Tuberculosis Research, US/MRC Centre for Molecular and Cellular Biology, Division of Molecular Biology and Human Genetics, Department of Biomedical Sciences, Faculty of Health Sciences, Stellenbosch University, Cape Town, South Africa,³ Institute of Thoracic Diseases, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil,⁴ Wadsworth Center, New York State Department of Health, Albany, New York,⁵ Public Health Research Institute, Newark, New Jersey,⁶ Unité de la Tuberculose et des Mycobactéries, Institut Pasteur de Guadeloupe, Guadeloupe,⁷ Unité de Génétique Mycobactérienne, Institut Pasteur, Paris, France,⁸ Departamento de Clínica Médica/Pneumologia, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil,⁹ GHESKIO Centers, Port-au-Prince, Haiti,¹⁰ National Institute of Public Health and the Environment, Bilthoven, The Netherlands,¹¹ Laboratoire des Mycobactéries, Institut Pasteur de Tunis, Tunisia,¹² Department of Anthropology, Graduate School and University Center, City University of New York/New York Consortium in Evolutionary Primatology, New York, New York,¹³ Servicio de Microbiología Clínica y Enfermedades Infecciosas, Hospital Gregorio Marañón, Madrid, Spain,¹⁴ CIBER Enfermedades Respiratorias, Madrid, Spain,¹⁵ Laboratory of Molecular Biology Applied to Mycobacteria, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil,¹⁶

Received 19 November 2007/ Returned for modification 27 December 2007/ Accepted 9 January 2008

The Latin American-Mediterranean (LAM) family of Mycobacterium tuberculosis is believed to be the cause of 15% of tuberculosis cases worldwide. Previously, we defined a prevalent sublineage of the LAM family in Brazil by a single characteristic genomic deletion designated RD^Rio. Using the Brazilian strains, we pinpoint an Ag85C¹⁰³ single nucleotide polymorphism (SNP) (screened by restriction fragment length polymorphism [RFLP] analysis) that correctly identified all LAM family strains. Importantly, all RD^Rio strains concomitantly possessed the RD174 deletion. These genetic signatures, along with a newly developed multiplex PCR for rapid differentiation between "wild-type" and RD^Rio strains, were then used to analyze an international collection of M. tuberculosis strains. RD^Rio M. tuberculosis was identified from four continents involving 11 countries. Phylogenetic analysis of the IS6110-RFLP patterns from representative RD^Rio and LAM strains from Brazil, along with all representative clusters from a South African database, confirmed their genetic relatedness and transcontinental transmission. The Ag85C¹⁰³ SNP RFLP, as compared to results obtained using a PCR method targeting a LAM-restricted IS6110 element, correctly identified 99.8% of LAM spoligotype strains. Together, these tests were more accurate than spoligotyping at categorizing strains with indefinable spoligotypes and segregated true LAM strains from those with convergent spoligotypes. The fact that RD^Rio strains were identified worldwide highlights the importance of this LAM family sublineage and suggests that this strain is a global threat that should be specifically targeted by public health resources. Our provision of simple and robust molecular methods will assist the evaluation of the LAM family and the RD^Rio sublineage.

Published ahead of print on 30 January 2008.

Supplemental material for this article may be found at http://jcm.asm.org/.

A.L.G. and R.C.H. contributed equally to this study.

This article has been cited by other articles:

• Lazzarini, L. C. O., Spindola, S. M., Bang, H., Gibson, A. L., Weisenberg, S., da Silva Carvalho, W., Augusto, C. J., Huard, R. C., Kritski, A. L., Ho, J. L. (2008). RDRio Mycobacterium tuberculosis Infection Is Associated with a Higher Frequency of Cavitary Pulmonary Disease. J. Clin. Microbiol. 46: 2175-2183 [Abstract] [Full Text]