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	Detection of mutations in rifampin-resistant mycobacterium tuberculosis by short oligonucleotide ligation assay on dna chips (solac)
	Zhang, XE; Deng, JY
刊名	Immobilisation of dna on chips ii
	2005
卷号	261页码:169-190
关键词	Enzyme-linked assay M. tuberculosis Mutation detection Rifampin resistance Short oligonucleotide ligation on dna chips (solac) T4 dna ligase
ISSN号	0340-1022
DOI	10.1007/128_004
通讯作者	Zhang, xe(zhangxe@mail.most.gov.cn)
英文摘要	A new approach, a short oligonucleotide ligation assay on dna chips (solac) is developed to detect dna mutations. the solac approach can be carried out through two experimental schemes: loss-of-signal (solac-los) and gain-of-signal (solac-gos). in both experimental schemes, probes with a disulfide modification on their terminuses are immobilized onto mercaptosflane derivatized glass slides through a thiol/clisulfide exchange reaction. in solac-los, the common probe is immobilized on the chip, and the allele-specific probe is used as the detecting probe; both probes are perfectly complementary to the wild-type target dna. after hybridization of sample dna with the immobilized common probe, t4 dna ligase is applied to ligate the common probe and to detect the probe. failure of ligation occurs if there is any mismatch between the sample dna and the detecting probe. this nick-containing hybrid conjugate cannot withstand denaturing and washing treatments, leading to the loss of signal, which indicates the presence of mutations in the target sample. theoretically, with one pair of probes (one common probe and one pentamer) all mutations (substitutions, insertions, and deletions) in the five-nucleoside region of the target dna can be detected. by contrast, in solac-gos, the solid phase is the array of allele-specific probes, which are designed to be complementary to all of the known mutations of the target region of the sample dna, while the common probes are detecting probes. after hybridization, ligation, and washing, the gain of signal is an indicator of the presence of mutations. for a five-base region of the target dna, basically sixteen allele-specific pentamers and just one common probe are needed to detect all possible mutations. in combination with an alkaline phosphatase reaction-linked assay, these two schemes have been used successfully for the identification of mutations in the rpob gene of mycobacterium tuberculosis from clinical isolates that show rifampin resistance (rif(r)). the advantages and disadvantages of the new approach are discussed.
WOS关键词	STRAND CONFORMATION POLYMORPHISM ; POLYMERASE-CHAIN-REACTION ; LINE PROBE ASSAY ; RAPID DETECTION ; SPECIES IDENTIFICATION ; MULTIPLEX DETECTION ; DRUG-RESISTANCE ; RPOB GENE ; LIGASE ; HYBRIDIZATION
WOS研究方向	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Chemistry
WOS类目	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Chemistry, Multidisciplinary
语种	英语
出版者	SPRINGER-VERLAG BERLIN
WOS记录号	WOS:000236941000008
内容类型	期刊论文
URI标识	http://www.corc.org.cn/handle/1471x/2375229
专题	武汉病毒研究所
通讯作者	Zhang, XE
作者单位	1.Chinese Acad Sci, State Key Lab Virol, Wuhan Inst Virol, Wuhan 430071, Peoples R China 2.Chinese Acad Sci, Inst Biophys, State Key Lab Biomacromol, Wuhan 430071, Peoples R China
推荐引用方式 GB/T 7714	Zhang, XE,Deng, JY. Detection of mutations in rifampin-resistant mycobacterium tuberculosis by short oligonucleotide ligation assay on dna chips (solac)[J]. Immobilisation of dna on chips ii,2005,261:169-190.
APA	Zhang, XE,&Deng, JY.(2005).Detection of mutations in rifampin-resistant mycobacterium tuberculosis by short oligonucleotide ligation assay on dna chips (solac).Immobilisation of dna on chips ii,261,169-190.
MLA	Zhang, XE,et al."Detection of mutations in rifampin-resistant mycobacterium tuberculosis by short oligonucleotide ligation assay on dna chips (solac)".Immobilisation of dna on chips ii 261(2005):169-190.

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