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dc.contributor.authorHuang, Qiuying
dc.contributor.authorZheng, Linlin
dc.contributor.authorZhu, Yumei
dc.contributor.authorZhang, Jiafeng
dc.contributor.authorWen, Huixin
dc.contributor.authorHuang, Jianwei
dc.contributor.authorNiu, Jianjun
dc.contributor.authorZhao, Xilin
dc.contributor.authorLi, Qingge
dc.contributor.author李庆阁
dc.date.accessioned2012-03-22T11:52:29Z
dc.date.available2012-03-22T11:52:29Z
dc.date.issued2011
dc.identifier.citationPLOS ONE 2010 vol. 6 no. e16033zh_CN
dc.identifier.issn1932-6203
dc.identifier.uriWOS:000286516500023
dc.identifier.urihttp://dx.doi.org/doi:10.1371/journal.pone.0016033
dc.identifier.urihttps://dspace.xmu.edu.cn/handle/2288/11843
dc.description.abstractThe target volume of multiplex real-time PCR assays is limited by the number of fluorescent dyes available and the number of fluorescence acquisition channels present in the PCR instrument. We hereby explored a probe labeling strategy that significantly increased the target volume of real-time PCR detection in one reaction. The labeling paradigm, termed "Multicolor Combinatorial Probe Coding'' (MCPC), uses a limited number (n) of differently colored fluorophores in various combinations to label each probe, enabling one of 2(n)-1 genetic targets to be detected in one reaction. The proof-of-principle of MCPC was validated by identification of one of each possible 15 human papillomavirus types, which is the maximum target number theoretically detectable by MCPC with a 4-color channel instrument, in one reaction. MCPC was then improved from a one-primer-pair setting to a multiple-primer-pair format through Homo-Tag Assisted Non-Dimer (HAND) system to allow multiple primer pairs to be included in one reaction. This improvement was demonstrated via identification of one of the possible 10 foodborne pathogen candidates with 10 pairs of primers included in one reaction, which had limit of detection equivalent to the uniplex PCR. MCPC was further explored in detecting combined genotypes of five beta-globin gene mutations where multiple targets were co-amplified. MCPC strategy could expand the scope of real-time PCR assays in applications which are unachievable by current labeling strategy.zh_CN
dc.description.sponsorshipNational Nature Science Foundation of China[30371366]; Science Research Foundation of Ministry of Health; United Fujian Provincial Health and Education, People's Republic of China[WKJ2005-2-002, WKJ2008-2-39]zh_CN
dc.language.isoenzh_CN
dc.publisherPUBLIC LIBRARY SCIENCEzh_CN
dc.titleMulticolor Combinatorial Probe Coding for Real-Time PCRzh_CN
dc.typeArticlezh_CN


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