PathogenMip Assay: A Multiplex Pathogen Detection Assay

被引：19

作者：

Akhras, Michael S. ^{[1
,2
]}

Thiyagarajan, Sreedevi ^{[1
]}

Villablanca, Andrea C. ^{[1
]}

Davis, Ronald W. ^{[1
]}

Nyren, Pal ^{[2
]}

Pourmand, Nader ^{[1
]}

机构：

[1] Stanford Univ, Stanford Genome Technol Ctr, Palo Alto, CA 94304 USA

[2] Royal Inst Technol, Dept Biotechnol, Stockholm, Sweden

来源：

PLOS ONE | 2007年 / 2卷 / 02期

基金：

美国国家卫生研究院;

关键词：

D O I：

10.1371/journal.pone.0000223

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：

The Molecular Inversion Probe (MIP) assay has been previously applied to a large-scale human SNP detection. Here we describe the PathogenMip Assay, a complete protocol for probe production and applied approaches to pathogen detection. We have demonstrated the utility of this assay with an initial set of 24 probes targeting the most clinically relevant HPV genotypes associated with cervical cancer progression. Probe construction was based on a novel, cost-effective, ligase-based protocol. The assay was validated by performing pyrosequencing and Microarray chip detection in parallel experiments. HPV plasmids were used to validate sensitivity and selectivity of the assay. In addition, 20 genomic DNA extracts from primary tumors were genotyped with the PathogenMip Assay results and were in 100% agreement with conventional sequencing using an L1-based HPV genotyping protocol. The PathogenMip Assay is a widely accessible protocol for producing and using highly discriminating probes, with experimentally validated results in pathogen genotyping, which could potentially be applied to the detection and characterization of any microbe.

引用

页数：11

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