A vertical flow paper-microarray assay with isothermal DNA amplification for detection of Neisseria meningitidis

TALANTA

卷:183

Article

Rivas, Lourdes¹  Reutersward, Philippa¹  Rasti, Reza^2,3  Herrmann, Bjorn⁴  Martensson, Andreas⁵  Alfven, Tobias^2,3  Gantelius, Jesper^1,2  Andersson-Svahn, Helene¹ 

[1] KTH Royal Inst Technol, Sci Life Lab, Div Prote & Nanobiotechnol, Stockholm, Sweden

[2] Karolinska Inst, Dept Publ Hlth Sci, Stockholm, Sweden

[3] South Gen Hosp, Sachs Children & Youth Hosp, Stockholm, Sweden

[4] Uppsala Univ, Dept Med Sci, Sect Clin Bacteriol, Uppsala, Sweden

[5] Uppsala Univ, IMCH, Dept Womens & Childrens Hlth, Uppsala, Sweden

关键词: Microarray;    Paper-based biosensor;    Point-of-care;    Gold nanoparticles;    Neisseria meningitidis;   

DOI  :  10.1016/j.talanta.2018.02.070

来源: Elsevier

【 摘 要 】

Paper-based biosensors offer a promising technology to be used at the point of care, enabled by good performance, convenience and low-cost. In this article, we describe a colorimetric vertical-flow DNA microarray (DNAVFM) that takes advantage of the screening capability of DNA microarrays in a paper format together with isothermal amplification by means of Recombinase Polymerase Amplification (RPA). Different assay parameters such as hybridization buffer, flow rate, printing buffer and capture probe concentration were optimized. A limit of detection (LOD) of 4.4 nM was achieved as determined by tabletop scanning. The DNA-VFM was applied as a proof of concept for detection of Neisseria meningitidis, a primary cause of bacterial meningitis. The LOD was determined to be between 38 and 2.1 x 10(6) copies/VFMassay, depending on the choice of DNA capture probes. The presented approach provides multiplex capabilities of DNA microarrays in a paper-based format for future point-of-care applications.

【 授权许可】

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