Ultrasensitive, rapid and inexpensive detection of DNA using paper based lateral flow assay.
Ultrasensitive, rapid and inexpensive detection of DNA using paper based lateral flow assay.
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DOI：
10.1038/srep37732
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发表时间：
2016-11-25
影响因子：
4.6
通讯作者：
O Sullivan CK
中科院分区：
文献类型：
--
作者：
Jauset-Rubio M;Svobodová M;Mairal T;McNeil C;Keegan N;Saeed A;Abbas MN;El-Shahawi MS;Bashammakh AS;Alyoubi AO;O Sullivan CK
在需要时使用的灵敏、特异、快速、廉价和易于使用的核酸测试对于伴随诊断必不可少的新兴个性化医疗领域以及在低资源环境中的应用至关重要。在这里，我们报告了一种用于直接检测等温扩增DNA的护理点核酸侧向流动试验的发展。重组酶聚合酶扩增方法略有修改，使用了拖尾引物，产生了一个两侧有两个单链DNA尾巴的双链扩增子。这种拖尾扩增子通过与表面固定的寡核苷酸捕获探针和金纳米颗粒标记的报告探针杂交来促进检测。检测下限为1 × 10−11 M(190 amol)，相当于8.67 × 105拷贝的DNA，整个分析，包括扩增和检测，在37 °C的恒定温度下，在不到15 分钟的时间内完成。尾随的引物的使用消除了对半抗原标记和随后捕获抗体和报告抗体的使用，同时也避免了为产生单链DNA而进行的任何扩增后处理，从而提供了一种能够在需要的点容易地找到应用的分析方法。
Sensitive, specific, rapid, inexpensive and easy-to-use nucleic acid tests for use at the point-of-need are critical for the emerging field of personalised medicine for which companion diagnostics are essential, as well as for application in low resource settings. Here we report on the development of a point-of-care nucleic acid lateral flow test for the direct detection of isothermally amplified DNA. The recombinase polymerase amplification method is modified slightly to use tailed primers, resulting in an amplicon with a duplex flanked by two single stranded DNA tails. This tailed amplicon facilitates detection via hybridisation to a surface immobilised oligonucleotide capture probe and a gold nanoparticle labelled reporter probe. A detection limit of 1 × 10−11 M (190 amol), equivalent to 8.67 × 105 copies of DNA was achieved, with the entire assay, both amplification and detection, being completed in less than 15 minutes at a constant temperature of 37 °C. The use of the tailed primers obviates the need for hapten labelling and consequent use of capture and reporter antibodies, whilst also avoiding the need for any post-amplification processing for the generation of single stranded DNA, thus presenting an assay that can facilely find application at the point of need.
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影响因子：
7.4
作者：
Crannell, Zachary Austin;Castellanos-Gonzalez, Alejandro;Irani, Ayesha;Rohrman, Brittany;White, Arthur Clinton;Richards-Kortum, Rebecca
通讯作者：
Richards-Kortum, Rebecca
影响因子：
4.9
作者：
Aveyard, Jenny;Mehrabi, Maryam;Wilson, Robert
通讯作者：
Wilson, Robert
影响因子：
7.4
作者：
Crannell, Zachary;Castellanos-Gonzalez, Alejandro;Richards-Kortum, Rebecca
通讯作者：
Richards-Kortum, Rebecca
影响因子：
9.4
作者：
Fong, WK;Modrusan, Z;Bekkaoui, F
通讯作者：
Bekkaoui, F
影响因子：
6.4
作者：
Boyle DS;Lehman DA;Lillis L;Peterson D;Singhal M;Armes N;Parker M;Piepenburg O;Overbaugh J
通讯作者：
Overbaugh J