Phosphorothioated Primers Lead to Loop-Mediated Isothermal Amplification at Low Temperatures

被引：83

作者：

Cai, Sheng ^{[1
]}

Jung, Cheulhee ^{[2
]}

Bhadra, Sanchita ^{[3
]}

Ellington, Andrew D. ^{[3
]}

机构：

[1] Zhejiang Univ, Lab Pharmaceut Anal & Drug Metab, Zhejiang Prov Key Lab Anticanc Drug Res, Hangzhou 310058, Zhejiang, Peoples R China

[2] Korea Univ, Coll Life Sci & Biotechnol, Div Biotechnol, Seoul 02841, South Korea

[3] Univ Texas Austin, Dept Mol Biosci, Inst Cellular & Mol Biol, Austin, TX 78712 USA

来源：

ANALYTICAL CHEMISTRY | 2018年 / 90卷 / 14期

基金：

美国国家航空航天局;

关键词：

SINGLE-STRANDED-DNA; IN-VITRO AMPLIFICATION; BINDING PROTEIN; LAMP; ASSAY; REPLICATION; MAGNESIUM; IONS; SPP; UREA;

D O I：

10.1021/acs.analchem.8b02062

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Loop-mediated isothermal amplification (LAMP) is an extremely powerful tool for the detection of nucleic acids with high sensitivity and specificity. However, LAMP shows optimal performance at around 65 degrees C, which limits applications in point-of-care-testing (POCT). Here, we have developed a version of LAMP that uses phosphorothioated primers (PS-LAMP) to enable more efficient hairpin formation and extension at the termini of growing concatamers, and that therefore works at much lower temperatures. By including additional factors such as chaotropes (urea) and single-stranded DNA binding protein (SSB), the sensitivities and selectivities for amplicon detection with PS-LAMP at 40 degrees C were comparable with a regular LAMP reaction at 65 degrees C.

引用

页码：8290 / 8294

页数：5

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