Rapid detection of Mycobacterium tuberculosis Beijing genotype strains by real-time PCR

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dc.contributor.authorHillemann D.
dc.contributor.authorWarren R.
dc.contributor.authorKubica T.
dc.contributor.authorRusch-Gerdes S.
dc.contributor.authorNiemann S.
dc.date.accessioned2011-05-15T15:56:56Z
dc.date.available2011-05-15T15:56:56Z
dc.date.issued2006
dc.description.abstractMycobacterium tuberculosis strains of the Beijing genotype were first identified in China and neighboring countries and have attracted special attention due to their global emergence and association with drug resistance. To further analyze the spread and special characteristics of Beijing genotype strains, accurate, rapid and sensitive methods that overcome the drawbacks of the classical methods such as IS6110 DNA fingerprinting or spoligotyping for the identification of strains of this genotype are needed. Based on the nucleotide sequences of M. tuberculosis SAWC0780 and H37Rv, primers and fluorogenic 5′ nuclease (TaqMan) probes for real-time PCR assays specific for Beijing and non-Beijing strains, respectively, were designed. The detection limits for the real-time PCR assays were about 5 and 10 copies of chromosomal DNA, respectively. In mixtures of Beijing and non-Beijing DNA, a multiplex assay was able to detect (i) one copy of Beijing DNA in approximately 1,000 copies of non-Beijing DNA and (ii) one copy of non-Beijing DNA in approximately 2,000 copies of Beijing DNA. In a blinded analysis of a collection of 103 multidrug-resistant strains isolated in Germany in 2001, all 62 Beijing and all 41 non-Beijing strains were correctly identified. In conclusion, the real-time assay allows for the rapid and specific detection of Beijing and non-Beijing strains. The major advantages of this test in comparison to other methods used for the identification of Beijing strains are its simplicity and sensitivity and the fact that amplification and detection occur within one reaction tube. Copyright © 2006, American Society for Microbiology. All Rights Reserved.
dc.description.versionArticle
dc.identifier.citationJournal of Clinical Microbiology
dc.identifier.citation44
dc.identifier.citation2
dc.identifier.issn951137
dc.identifier.other10.1128/JCM.44.2.302-306.2006
dc.identifier.urihttp://hdl.handle.net/10019.1/10119
dc.subjectbacterial DNA
dc.subjectchromosome deoxyribonucleic acid
dc.subjectnuclease
dc.subjectprimer DNA
dc.subjectTaq polymerase
dc.subjectunclassified drug
dc.subjectaccuracy
dc.subjectarticle
dc.subjectbacterial chromosome
dc.subjectbacterial strain
dc.subjectbacterium detection
dc.subjectbacterium isolate
dc.subjectbacterium isolation
dc.subjectDNA determination
dc.subjectDNA fingerprinting
dc.subjectgenotype
dc.subjectmultidrug resistance
dc.subjectmultiplex polymerase chain reaction
dc.subjectMycobacterium tuberculosis
dc.subjectnonhuman
dc.subjectnucleotide sequence
dc.subjectpriority journal
dc.subjectreal time polymerase chain reaction
dc.subjectBacterial Typing Techniques
dc.subjectChina
dc.subjectDNA Fingerprinting
dc.subjectDNA Probes
dc.subjectDNA Transposable Elements
dc.subjectDNA, Bacterial
dc.subjectGenotype
dc.subjectGermany
dc.subjectHumans
dc.subjectMycobacterium tuberculosis
dc.subjectPolymerase Chain Reaction
dc.subjectTime Factors
dc.subjectTuberculosis, Multidrug-Resistant
dc.subjectTuberculosis, Pulmonary
dc.subjectBacteria (microorganisms)
dc.subjectMycobacterium tuberculosis
dc.titleRapid detection of Mycobacterium tuberculosis Beijing genotype strains by real-time PCR
dc.typeArticle
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