Connector Inversion Probe Technology: A Powerful One-Primer Multiplex DNA Amplification System for Numerous Scientific Applications

被引:20
作者
Akhras, Michael S. [1 ,2 ]
Unemo, Magnus [3 ]
Thiyagarajan, Sreedevi [1 ]
Nyren, Pal [2 ]
Davis, Ronald W. [1 ]
Fire, Andrew Z. [4 ,5 ]
Pourmand, Nader [1 ,6 ]
机构
[1] Stanford Univ, Stanford Genome Technol Ctr, Palo Alto, CA 94304 USA
[2] Royal Inst Technol, Dept Biotechnol, Stockholm, Sweden
[3] Orebro Univ Hosp, Dept Clin Microbiol, Natl Reference Lab Pathogen Neisseria, Orebro, Sweden
[4] Stanford Univ, Dept Pathol, Sch Med, Stanford, CA 94305 USA
[5] Stanford Univ, Dept Genet, Sch Med, Stanford, CA 94305 USA
[6] Univ Calif Santa Cruz, Santa Cruz, CA 95064 USA
来源
PLOS ONE | 2007年 / 2卷 / 09期
基金
美国国家卫生研究院;
关键词
D O I
10.1371/journal.pone.0000915
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We combined components of a previous assay referred to as Molecular Inversion Probe (MIP) with a complete gap filling strategy, creating a versatile powerful one-primer multiplex amplification system. As a proof-of-concept, this novel method, which employs a Connector Inversion Probe (CIPer), was tested as a genetic tool for pathogen diagnosis, typing, and antibiotic resistance screening with two distinct systems: i) a conserved sequence primer system for genotyping Human Papillomavirus (HPV), a cancer-associated viral agent and ii) screening for antibiotic resistance mutations in the bacterial pathogen Neisseria gonorrhoeae. We also discuss future applications and advances of the CIPer technology such as integration with digital amplification and next-generation sequencing methods. Furthermore, we introduce the concept of two-dimension informational barcodes, i. e. "multiplex multiplexing padlocks'' (MMPs). For the readers' convenience, we also provide an on-line tutorial with user-interface software application CIP creator 1.0.1, for custom probe generation from virtually any new or established primer-pairs.
引用
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页数:6
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