Source:http://linkedlifedata.com/resource/pubmed/id/18722102
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
4
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pubmed:dateCreated |
2008-10-20
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pubmed:abstractText |
Detailed analyses of dense single nucleotide polymorphism (SNP) loci within rifampin-resistance determining region (RRDR) are very important for the early assessment of drug resistance of Mycobacterium tuberculosis. A strategy was developed here to specifically identify point mutations out of dense SNP loci by on-chip ligation of multiplexing probe-pairs (MPPs). A probe-pair combines a common probe with a discriminating probe which is covalently attached to a DNA chip. The common probe hybridizes to the discriminating probe via a unique "zip-code complement". The allele-specific part on the 3'-end of the discriminating probe becomes covalently ligated to the adjacent part on the 5'-end of the common probe if and only if a mutation is present. Thus upon zip-code recognition, the process of identifying a mutation of interest is entirely located into corresponding well on the chip. As a consequence, cross-reactions and biased competitive attachments to targets, both of which result from the presence of various multiplexing probes, are greatly minimized. Mutation detection was performed by direct visualization using enzyme-linked assay. The method was demonstrated with an initial set of 24 probe-pairs targeting 22 clinically meaningful mutations within an 81-bp RRDR. 130-bp fragments of the rpoB gene from 15 clinical isolates were identified and were in 100% agreement with results from independent sequencing.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Dec
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pubmed:issn |
1873-4235
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pubmed:author | |
pubmed:issnType |
Electronic
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pubmed:day |
1
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pubmed:volume |
24
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
818-24
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pubmed:dateRevised |
2009-7-14
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pubmed:meshHeading |
pubmed-meshheading:18722102-Bacterial Proteins,
pubmed-meshheading:18722102-DNA, Bacterial,
pubmed-meshheading:18722102-DNA Mutational Analysis,
pubmed-meshheading:18722102-DNA Probes,
pubmed-meshheading:18722102-Equipment Design,
pubmed-meshheading:18722102-Equipment Failure Analysis,
pubmed-meshheading:18722102-Mycobacterium tuberculosis,
pubmed-meshheading:18722102-Oligonucleotide Array Sequence Analysis,
pubmed-meshheading:18722102-Point Mutation,
pubmed-meshheading:18722102-Polymorphism, Single Nucleotide,
pubmed-meshheading:18722102-Reproducibility of Results,
pubmed-meshheading:18722102-Sensitivity and Specificity
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pubmed:year |
2008
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pubmed:articleTitle |
On-chip ligation of multiplexing probe-pairs for identifying point mutations out of dense SNP loci.
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pubmed:affiliation |
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Xiao Hong Shan, Wuchang, Wuhan 430071, PR China.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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