Molecular inversion probes for sensitive detection of Mycobacterium tuberculosis
Nucleic acid-based detection of Mycobacterium tuberculosis infections has the potential to improve the analysis of the tuberculosis epidemiology and patient care by increasing the specificity and sensitivity of diagnosis. One potential diagnostic sequence, the DR locus, is present in all isolates of...
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| Veröffentlicht in: | Journal of microbiological methods 2008, Vol.72 (1), p.60-66 |
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| creator | Novais, Rogerio C. Borsuk, Sibelle Dellagostin, Odir A. Thorstenson, Yvonne R. |
| description | Nucleic acid-based detection of
Mycobacterium tuberculosis infections has the potential to improve the analysis of the tuberculosis epidemiology and patient care by increasing the specificity and sensitivity of diagnosis. One potential diagnostic sequence, the DR locus, is present in all isolates of
M. tuberculosis complex bacteria. It encodes no known gene product but is useful for molecular typing of
M. tuberculosis because of its fortuitous absence in non-tuberculosis strains of mycobacteria. The DR locus contains a variable number of short direct repeats interspersed with non-repetitive spacers and is commonly used as a target for the spoligotyping method, a technique based on the detection of the presence or absence of distinct spacers between the repeats. In this study, we attempted to combine the specificity of molecular inversion probe (MIP) technology with the sensitivity of modified pyrosequencing readout in order to detect a short conserved 18 bp sequence included in DR locus in 25 isolates of
M. tuberculosis. Additional sensitivity was obtained by introducing modifications in pyrosequencing methodology, by these means we achieved to detect 500 fg of
M. tuberculosis DNA. |
| doi_str_mv | 10.1016/j.mimet.2007.10.009 |
| format | Article |
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Mycobacterium tuberculosis infections has the potential to improve the analysis of the tuberculosis epidemiology and patient care by increasing the specificity and sensitivity of diagnosis. One potential diagnostic sequence, the DR locus, is present in all isolates of
M. tuberculosis complex bacteria. It encodes no known gene product but is useful for molecular typing of
M. tuberculosis because of its fortuitous absence in non-tuberculosis strains of mycobacteria. The DR locus contains a variable number of short direct repeats interspersed with non-repetitive spacers and is commonly used as a target for the spoligotyping method, a technique based on the detection of the presence or absence of distinct spacers between the repeats. In this study, we attempted to combine the specificity of molecular inversion probe (MIP) technology with the sensitivity of modified pyrosequencing readout in order to detect a short conserved 18 bp sequence included in DR locus in 25 isolates of
M. tuberculosis. Additional sensitivity was obtained by introducing modifications in pyrosequencing methodology, by these means we achieved to detect 500 fg of
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Mycobacterium tuberculosis infections has the potential to improve the analysis of the tuberculosis epidemiology and patient care by increasing the specificity and sensitivity of diagnosis. One potential diagnostic sequence, the DR locus, is present in all isolates of
M. tuberculosis complex bacteria. It encodes no known gene product but is useful for molecular typing of
M. tuberculosis because of its fortuitous absence in non-tuberculosis strains of mycobacteria. The DR locus contains a variable number of short direct repeats interspersed with non-repetitive spacers and is commonly used as a target for the spoligotyping method, a technique based on the detection of the presence or absence of distinct spacers between the repeats. In this study, we attempted to combine the specificity of molecular inversion probe (MIP) technology with the sensitivity of modified pyrosequencing readout in order to detect a short conserved 18 bp sequence included in DR locus in 25 isolates of
M. tuberculosis. Additional sensitivity was obtained by introducing modifications in pyrosequencing methodology, by these means we achieved to detect 500 fg of
M. tuberculosis DNA.</description><subject>Bacterial Typing Techniques</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Diphosphates - metabolism</subject><subject>DNA, Bacterial - analysis</subject><subject>DNA, Bacterial - isolation & purification</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Inorganic Chemicals - metabolism</subject><subject>Microbiology</subject><subject>Molecular inversion probes</subject><subject>Molecular Probe Techniques</subject><subject>Molecular Probes</subject><subject>Molecular Sequence Data</subject><subject>Mycobacterium tuberculosis</subject><subject>Mycobacterium tuberculosis - classification</subject><subject>Mycobacterium tuberculosis - genetics</subject><subject>Mycobacterium tuberculosis - isolation & purification</subject><subject>Oligonucleotides - genetics</subject><subject>Pyrosequencing</subject><subject>Repetitive Sequences, Nucleic Acid - genetics</subject><subject>Sensitivity and Specificity</subject><subject>Sequence Analysis, DNA - methods</subject><subject>Tuberculosis</subject><subject>Tuberculosis - diagnosis</subject><subject>Tuberculosis - microbiology</subject><issn>0167-7012</issn><issn>1872-8359</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkEFL5TAQx4Mo-lb9BIL0orc-k6ZNmoMHkXVXUPTgPSTpBPJoG820D_z2m_oeeFtPAzO_-TPzI-SC0TWjTNxs1kMYYFpXlMrcWVOqDsiKtbIqW96oQ7LKlCwlZdUJ-YW4oZQ1vG6PyQlrKReirVfk9Tn24ObepCKMW0gY4li8p2gBCx9TgTBimMIWig4mcNMyjr54_nTRGjdBCvNQTLOFlEMiBjwjR970COf7ekreHn6_3f8tn17-PN7fPZWuZu1UNkI1RikOXtXMSCMAqOqEkJw2jFtLvXA1tbXIUNVK5pzwvJZWgmXKK35Krnex-daPGXDSQ0AHfW9GiDPq_LXMYeJHkKmmaphcEvkOdCkiJvD6PYXBpE_NqF6E643-Eq4X4UszC89bl_v42Q7Qfe_sDWfgag8YdKb3yYwu4DenlKhVs3C3Ow6ytG2ApNEFGB10IWXvuovhv4f8AzAkoFw</recordid><startdate>2008</startdate><enddate>2008</enddate><creator>Novais, Rogerio C.</creator><creator>Borsuk, Sibelle</creator><creator>Dellagostin, Odir A.</creator><creator>Thorstenson, Yvonne R.</creator><general>Elsevier B.V</general><general>Elsevier Science</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>2008</creationdate><title>Molecular inversion probes for sensitive detection of Mycobacterium tuberculosis</title><author>Novais, Rogerio C. ; Borsuk, Sibelle ; Dellagostin, Odir A. ; Thorstenson, Yvonne R.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c418t-5695a993ef941a7a6ee09d66730513bb0f6c40b46a992871cc6f347b7eb19f93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Bacterial Typing Techniques</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Diphosphates - metabolism</topic><topic>DNA, Bacterial - analysis</topic><topic>DNA, Bacterial - isolation & purification</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Inorganic Chemicals - metabolism</topic><topic>Microbiology</topic><topic>Molecular inversion probes</topic><topic>Molecular Probe Techniques</topic><topic>Molecular Probes</topic><topic>Molecular Sequence Data</topic><topic>Mycobacterium tuberculosis</topic><topic>Mycobacterium tuberculosis - classification</topic><topic>Mycobacterium tuberculosis - genetics</topic><topic>Mycobacterium tuberculosis - isolation & purification</topic><topic>Oligonucleotides - genetics</topic><topic>Pyrosequencing</topic><topic>Repetitive Sequences, Nucleic Acid - genetics</topic><topic>Sensitivity and Specificity</topic><topic>Sequence Analysis, DNA - methods</topic><topic>Tuberculosis</topic><topic>Tuberculosis - diagnosis</topic><topic>Tuberculosis - microbiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Novais, Rogerio C.</creatorcontrib><creatorcontrib>Borsuk, Sibelle</creatorcontrib><creatorcontrib>Dellagostin, Odir A.</creatorcontrib><creatorcontrib>Thorstenson, Yvonne R.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of microbiological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Novais, Rogerio C.</au><au>Borsuk, Sibelle</au><au>Dellagostin, Odir A.</au><au>Thorstenson, Yvonne R.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular inversion probes for sensitive detection of Mycobacterium tuberculosis</atitle><jtitle>Journal of microbiological methods</jtitle><addtitle>J Microbiol Methods</addtitle><date>2008</date><risdate>2008</risdate><volume>72</volume><issue>1</issue><spage>60</spage><epage>66</epage><pages>60-66</pages><issn>0167-7012</issn><eissn>1872-8359</eissn><coden>JMIMDQ</coden><abstract>Nucleic acid-based detection of
Mycobacterium tuberculosis infections has the potential to improve the analysis of the tuberculosis epidemiology and patient care by increasing the specificity and sensitivity of diagnosis. One potential diagnostic sequence, the DR locus, is present in all isolates of
M. tuberculosis complex bacteria. It encodes no known gene product but is useful for molecular typing of
M. tuberculosis because of its fortuitous absence in non-tuberculosis strains of mycobacteria. The DR locus contains a variable number of short direct repeats interspersed with non-repetitive spacers and is commonly used as a target for the spoligotyping method, a technique based on the detection of the presence or absence of distinct spacers between the repeats. In this study, we attempted to combine the specificity of molecular inversion probe (MIP) technology with the sensitivity of modified pyrosequencing readout in order to detect a short conserved 18 bp sequence included in DR locus in 25 isolates of
M. tuberculosis. Additional sensitivity was obtained by introducing modifications in pyrosequencing methodology, by these means we achieved to detect 500 fg of
M. tuberculosis DNA.</abstract><cop>Shannon</cop><pub>Elsevier B.V</pub><pmid>18036684</pmid><doi>10.1016/j.mimet.2007.10.009</doi><tpages>7</tpages></addata></record> |
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| subjects | Bacterial Typing Techniques Base Sequence Biological and medical sciences Diphosphates - metabolism DNA, Bacterial - analysis DNA, Bacterial - isolation & purification Fundamental and applied biological sciences. Psychology Humans Inorganic Chemicals - metabolism Microbiology Molecular inversion probes Molecular Probe Techniques Molecular Probes Molecular Sequence Data Mycobacterium tuberculosis Mycobacterium tuberculosis - classification Mycobacterium tuberculosis - genetics Mycobacterium tuberculosis - isolation & purification Oligonucleotides - genetics Pyrosequencing Repetitive Sequences, Nucleic Acid - genetics Sensitivity and Specificity Sequence Analysis, DNA - methods Tuberculosis Tuberculosis - diagnosis Tuberculosis - microbiology |
| title | Molecular inversion probes for sensitive detection of Mycobacterium tuberculosis |
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