Stanford Genome Technology Center, Stanford University, Palo Alto, California, United States of America.
PLoS One. 2007 Sep 19;2(9):e915. doi: 10.1371/journal.pone.0000915.
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.
我们将先前的分子反转探针（MIP）检测法与完整的缺口填充策略相结合，创建了一个通用的强大单引物多重扩增系统。作为概念验证，这种新颖的方法采用连接器反转探针（CIPer），作为病原体诊断、分型和抗生素耐药性筛选的遗传工具进行了测试，该方法适用于两个不同的系统：i）用于 HPV（人乳头瘤病毒）分型的保守序列引物系统，HPV 是一种与癌症相关的病毒；ii）筛选淋病奈瑟菌（细菌病原体）中的抗生素耐药性突变。我们还讨论了 CIPer 技术的未来应用和进展，例如与数字扩增和下一代测序方法的整合。此外，我们引入了二维信息条码的概念，即“多重多重锁（multiplex multiplexing padlocks）”。为了方便读者，我们还提供了一个在线教程，其中包含用户界面软件应用程序 CIP creator 1.0.1，可用于从几乎任何新的或现有的引物对生成自定义探针。
