Characterization of Mycobacterium tuberculosis Isolates from Patients in Houston, Texas, by Spoligotyping

ABSTRACT Mycobacterium tuberculosis isolates (n= 1,429) from 1,283 patients collected as part of an ongoing population-based tuberculosis epidemiology study in Houston, Texas, were analyzed by spoligotyping and IS6110 profiling. The isolates were also assigned to one of three major genetic groups on the basis of nucleotide polymorphisms located at codons 463 and 95 in the genes (katG and gyrA) encoding catalase-peroxidase and the A subunit of DNA gyrase, respectively. A total of 225 spoligotypes were identified in the 1,429 isolates. There were 54 spoligotypes identified among 713 isolates (n= 623 patients) assigned to 73 IS6110 clusters. In addition, among 716 isolates (n = 660 patients) with unique IS6110 profiles, 200 spoligotypes were identified. No changes were observed either in the IS6110 profile or in the spoligotype for the 281 isolates collected sequentially from 133 patients. Five instances in which isolates with slightly different spoligotypes had the same IS6110 profile were identified, suggesting that in rare cases isolates with different spoligotypes can be clonally related. Spoligotypes correlated extremely well with major genetic group designations. Only three very similar spoligotypes were shared by isolates from genetic groups 2 and 3, and none was shared by group 1 and group 2 organisms or by group 1 and group 3 organisms. All organisms belonging to genetic groups 2 and 3 failed to hybridize with spacer probes 33 to 36. Taken together, the results support the existence of three distinct genetic groups of M. tuberculosis organisms and provide new information about the relationship between IS6110 profiles, spoligotypes, and major genetic groups of M. tuberculosis.

[1]  B. Gicquel,et al.  Evaluation of spoligotyping in a study of the transmission of Mycobacterium tuberculosis , 1997, Journal of clinical microbiology.

[2]  D. van Soolingen,et al.  Usefulness of Spoligotyping in Molecular Epidemiology of Mycobacterium bovis-Related Infections in South America , 1999, Journal of Clinical Microbiology.

[3]  R. Reves,et al.  Spread of strain W, a highly drug-resistant strain of Mycobacterium tuberculosis, across the United States. , 1999, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[4]  F. Baquero,et al.  Genetic Characterization of Multidrug-ResistantMycobacterium bovis Strains from a Hospital Outbreak Involving Human Immunodeficiency Virus-Positive Patients , 1998, Journal of Clinical Microbiology.

[5]  R. Auckenthaler,et al.  Mycobacterium canettii, the smooth variant of M. tuberculosis, isolated from a Swiss patient exposed in Africa. , 1998, Emerging infectious diseases.

[6]  T. Whittam,et al.  Restricted structural gene polymorphism in the Mycobacterium tuberculosis complex indicates evolutionarily recent global dissemination. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[7]  D. van Soolingen,et al.  Diagnosis of Mycobacterium microtiInfections among Humans by Using Novel Genetic Markers , 1998, Journal of Clinical Microbiology.

[8]  J. Buring,et al.  Epidemiology in Medicine , 1987 .

[9]  D van Soolingen,et al.  Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology , 1997, Journal of clinical microbiology.

[10]  H. Miörner,et al.  Usefulness of Spoligotyping To Discriminate IS6110Low-Copy-Number Mycobacterium tuberculosis Complex Strains Cultured in Denmark , 1999, Journal of Clinical Microbiology.

[11]  Jeffrey R. Driscoll,et al.  DNA Typing of a Nonviable Culture of Mycobacterium tuberculosis in a Homeless Shelter Outbreak , 1999, Journal of Clinical Microbiology.

[12]  J. T. Crawford,et al.  Strain identification of Mycobacterium tuberculosis by DNA fingerprinting: recommendations for a standardized methodology , 1993, Journal of clinical microbiology.

[13]  J T Douglas,et al.  Predominance of a single genotype of Mycobacterium tuberculosis in countries of east Asia , 1995, Journal of clinical microbiology.

[14]  D. van Soolingen,et al.  Mycobacterium microti: More Widespread than Previously Thought , 1998, Journal of Clinical Microbiology.

[15]  D. Young,et al.  Differentiation of Mycobacterium tuberculosis isolates by spoligotyping and IS6110 restriction fragment length polymorphism , 1997, Journal of clinical microbiology.

[16]  T. Ellis,et al.  Evaluation of Four DNA Typing Techniques in Epidemiological Investigations of Bovine Tuberculosis , 1998, Journal of Clinical Microbiology.

[17]  D. Young,et al.  Genotypic analysis of Mycobacterium tuberculosis from medieval human remains. , 1999, Microbiology.

[18]  J. V. van Embden,et al.  Identification by spoligotyping of a caprine genotype in Mycobacterium bovis strains causing human tuberculosis , 1997, Journal of clinical microbiology.

[19]  Nalin Rastogi,et al.  Spoligotyping Followed by Double-Repetitive-Element PCR as Rapid Alternative to IS6110 Fingerprinting for Epidemiological Studies of Tuberculosis , 1998, Journal of Clinical Microbiology.

[20]  M. Domingo,et al.  Spacer oligonucleotide typing of Mycobacterium bovis strains from cattle and other animals: a tool for studying epidemiology of tuberculosis , 1996, Journal of clinical microbiology.

[21]  S. Goss,et al.  Evaluation of Strategies for Molecular Fingerprinting for Use in the Routine Work of a Mycobacterium Reference Unit , 1998, Journal of Clinical Microbiology.

[22]  J. Bates,et al.  Diversity of DNA Fingerprints ofMycobacterium tuberculosis Isolates in the United States , 1998, Journal of Clinical Microbiology.

[23]  P. Groenen,et al.  Nature of DNA polymorphism in the direct repeat cluster of Mycobacterium tuberculosis; application for strain differentiation by a novel typing method , 1993, Molecular microbiology.

[24]  R. Frothingham,et al.  Comparison of Methods Based on Different Molecular Epidemiological Markers for Typing of Mycobacterium tuberculosis Complex Strains: Interlaboratory Study of Discriminatory Power and Reproducibility , 1999, Journal of Clinical Microbiology.

[25]  R. Quesada,et al.  Espinosa de los monteros , 2000 .

[26]  Follow up of Mycobacterium tuberculosis transmission in the French West Indies by IS6110-DNA fingerprinting and DR-based spoligotyping. , 1998, FEMS immunology and medical microbiology.

[27]  J. T. Crawford,et al.  Origin and interstate spread of a New York City multidrug-resistant Mycobacterium tuberculosis clone family. , 1996, JAMA.

[28]  J. V. van Embden,et al.  Retrospective Analysis of the Beijing Family ofMycobacterium tuberculosis in Preserved Lung Tissues , 1999, Journal of Clinical Microbiology.

[29]  E. Legrand,et al.  Tuberculosis in the Caribbean: using spacer oligonucleotide typing to understand strain origin and transmission. , 1999, Emerging infectious diseases.

[30]  S. Niemann,et al.  Stability of Mycobacterium tuberculosis IS6110 restriction fragment length polymorphism patterns and spoligotypes determined by analyzing serial isolates from patients with drug-resistant tuberculosis. , 1999, Journal of clinical microbiology.