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首页> 外文期刊>Science and Technology of Advanced Materials >Peptide nucleic acid modified magnetic beads for intercalator based electrochemical detection of DNA hybridization
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Peptide nucleic acid modified magnetic beads for intercalator based electrochemical detection of DNA hybridization

机译：多肽核酸修饰的磁珠用于基于嵌入剂的DNA杂交电化学检测

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摘要

The specific binding of peptide nucleic acid (PNA) to its complementary DNA target is combined with magnetic separation to enable discrimination against single nucleotide polymorphisms (SNP). PNA probes with biotin label at 5'-end were attached to strepavidin coated superparamagnetic iron oxide beads. PNA modified beads were then challenged with non-complementary, SNP containing and perfect-match DNA targets. PNA probe showed no affinity towards non-complementary DNA. The non-specific binding of SNP containing DNA target was suppressed by the washing step of the beads by using sodium dodecylsulfate in blank buffer solution. Then, an electro-active intercalator, 7-dimethyl-amino-1,2-benzophenoxazinium salt (Meldola's blue, MDB) was introduced to the beads. MDB intercalated between the double-helix of the hybrid molecules on the beads. After removing the excessive MDB, the beads were collected from the solution by immersing a biotin modified carbon paste electrode into the solution. Specific hybridization between PNA probe and DNA target was determined by monitoring the voltammetric peak of MDB. Numerous factors affecting the MDB signal, such as target DNA concentration, intercalator concentration and accumulation time were investigated. MDB signal indicated a detection limit of 2 pM in connection with 20 min hybridization time.
机译：肽核酸（PNA）与其互补DNA靶标的特异性结合与磁分离相结合，可以区分单核苷酸多态性（SNP）。将在5'端带有生物素标记的PNA探针连接到链霉亲和素包被的超顺磁性氧化铁珠上。然后用非互补的，含有SNP和完美匹配的DNA靶标攻击PNA修饰的珠子。 PNA探针对非互补DNA无亲和力。通过在空白缓冲溶液中使用十二烷基硫酸钠洗涤珠子，可以抑制含有SNP的DNA靶标的非特异性结合。然后，将电活性嵌入剂7-二甲基-氨基-1,2-苯并苯恶嗪盐（Meldola's blue，MDB）引入微珠。 MDB插在珠子上的杂化分子的双螺旋之间。去除过量的MDB后，通过将生物素修饰的碳糊电极浸入溶液中，从溶液中收集珠子。通过监测MDB的伏安峰确定PNA探针和DNA靶之间的特异性杂交。研究了影响MDB信号的许多因素，例如目标DNA浓度，嵌入剂浓度和累积时间。 MDB信号表明与20分钟杂交时间相关的2 pM检测极限。

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