The Sequencing Bead Array (SBA), a Next-Generation Digital Suspension Array

被引:5
作者
Akhras, Michael S. [1 ]
Pettersson, Erik [1 ]
Diamond, Lisa [1 ]
Unemo, Magnus [2 ]
Okamoto, Jennifer [3 ,4 ]
Davis, Ronald W. [1 ]
Pourmand, Nader [5 ]
机构
[1] Stanford Univ, Stanford Genome Technol Ctr, Palo Alto, CA 94304 USA
[2] Orebro Univ Hosp, World Hlth Org Collaborating Ctr Gonorrhoea & Oth, Swedish Reference Lab Pathogen Neisseria, Dept Lab Med, Orebro, Sweden
[3] Stanford Univ, Dept Bioengn, Stanford, CA 94305 USA
[4] Stanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
[5] Univ Calif Santa Cruz, Dept Biomol Engn, Santa Cruz, CA 95064 USA
基金
美国国家卫生研究院;
关键词
DIRECT ELECTRICAL DETECTION; PADLOCK PROBES; IN-SITU; DNA; AMPLIFICATION; TECHNOLOGY; ASSAY; CLASSIFICATION; MOLECULES; PRIMERS;
D O I
10.1371/journal.pone.0076696
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
Here we describe the novel Sequencing Bead Array (SBA), a complete assay for molecular diagnostics and typing applications. SBA is a digital suspension array using Next-Generation Sequencing (NGS), to replace conventional optical readout platforms. The technology allows for reducing the number of instruments required in a laboratory setting, where the same NGS instrument could be employed from whole-genome and targeted sequencing to SBA broad-range biomarker detection and genotyping. As proof-of-concept, a model assay was designed that could distinguish ten Human Papillomavirus (HPV) genotypes associated with cervical cancer progression. SBA was used to genotype 20 cervical tumor samples and, when compared with amplicon pyrosequencing, was able to detect two additional co-infections due to increased sensitivity. We also introduce in-house software Sphix, enabling easy accessibility and interpretation of results. The technology offers a multi-parallel, rapid, robust, and scalable system that is readily adaptable for a multitude of microarray diagnostic and typing applications, e. g. genetic signatures, single nucleotide polymorphisms (SNPs), structural variations, and immunoassays. SBA has the potential to dramatically change the way we perform probe-based applications, and allow for a smooth transition towards the technology offered by genomic sequencing.
引用
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页数:13
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