Publication:
Optimized molecular resolution of cross-contamination alerts in clinical mycobacteriology laboratories

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Date

2008-02-14

Authors

Martín, Ana
Herranz, Marta
Martínez Lirola, Miguel
Fernández Fernández, Rosa
Bouza, Emilio
García de Viedma, Darío

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BMC Microbiology
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Abstract

BACKGROUND. The phenomenon of misdiagnosing tuberculosis (TB) by laboratory cross-contamination when culturing Mycobacterium tuberculosis (MTB) has been widely reported and it has an obvious clinical, therapeutic and social impact. The final confirmation of a cross-contamination event requires the molecular identification of the same MTB strain cultured from both the potential source of the contamination and from the false-positive candidate. The molecular tool usually applied in this context is IS6110-RFLP which takes a long time to provide an answer, usually longer than is acceptable for microbiologists and clinicians to make decisions. Our purpose in this study is to evaluate a novel PCR-based method, MIRU-VNTR as an alternative to assure a rapid and optimized analysis of cross-contamination alerts. RESULTS. MIRU-VNTR was prospectively compared with IS6110-RFLP for clarifying 19 alerts of false positivity from other laboratories. MIRU-VNTR highly correlated with IS6110-RFLP, reduced the response time by 27 days and clarified six alerts unresolved by RFLP. Additionally, MIRU-VNTR revealed complex situations such as contamination events involving polyclonal isolates and a false-positive case due to the simultaneous cross-contamination from two independent sources. CONCLUSION. Unlike standard RFLP-based genotyping, MIRU-VNTR i) could help reduce the impact of a false positive diagnosis of TB, ii) increased the number of events that could be solved and iii) revealed the complexity of some cross-contamination events that could not be dissected by IS6110-RFLP.

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INDAL-TB group: MI Sánchez, MC Rogado, T Cabezas, W SánchezYebra, J Martínez, MA Lucerna, P Barroso, I Cabeza- Barrera, LF Díez, M Rodríguez, M Escámez, P Marín, A Lazo, J Gamir, J Vázquez, C Gutiérrez, A Reyes and T Peñafiel. Journal Article; Research Support, Non-U.S. Gov't;

MeSH Terms

Medical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Diagnostic Errors::False Positive Reactions
Medical Subject Headings::Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans
Medical Subject Headings::Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Molecular Structure::Base Sequence::Repetitive Sequences, Nucleic Acid::Interspersed Repetitive Sequences
Medical Subject Headings::Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Molecular Structure::Base Sequence::Repetitive Sequences, Nucleic Acid::Tandem Repeat Sequences::Minisatellite Repeats
Medical Subject Headings::Organisms::Bacteria::Gram-Positive Bacteria::Actinobacteria::Actinomycetales::Mycobacteriaceae::Mycobacterium::Mycobacterium tuberculosis
Medical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Nucleic Acid Amplification Techniques::Polymerase Chain Reaction
Medical Subject Headings::Phenomena and Processes::Genetic Phenomena::Genetic Variation::Polymorphism, Genetic::Polymorphism, Restriction Fragment Length
Medical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Epidemiologic Methods::Epidemiologic Study Characteristics as Topic::Epidemiologic Studies::Cohort Studies::Longitudinal Studies::Prospective Studies
Medical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Clinical Laboratory Techniques::Specimen Handling
Medical Subject Headings::Diseases::Bacterial Infections and Mycoses::Bacterial Infections::Gram-Positive Bacterial Infections::Actinomycetales Infections::Mycobacterium Infections::Tuberculosis
Medical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Diagnostic Errors

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Keywords

Tuberculosis, Errores Diagnósticos, Estudios Prospectivos, Reacciones Falso Positivas

Citation

Martín A, Herranz M, Martínez Lirola M, Fernández Fernández R, Bouza E, García de Viedma D. Optimized molecular resolution of cross-contamination alerts in clinical mycobacteriology laboratories. BMC Microbiol.; 8:30