Colorimetric Detection of Polynucleotides on Polypropylene Slices
Colorimetric Detection of Polynucleotides on Polypropylene Slices
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DOI：
10.2116/analsci.20.461
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发表时间：
2004-03
影响因子：
1.6
通讯作者：
Libo Nie;Jian-xin Tang;Huishi Guo;Hong Chen;Pengfeng Xiao;N. He
中科院分区：
文献类型：
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作者：
Libo Nie;Jian-xin Tang;Huishi Guo;Hong Chen;Pengfeng Xiao;N. He
金-银染色法（GLSS）作为一种比色检测方法，具有无放射性、荧光不猝灭、分析设备简单等优点，近年来受到越来越多的关注。采用GLSS法对聚丙烯（PP）切片上原位合成的DNA阵列进行比色检测。本文采用简单的等离子体处理方法在聚丙烯薄片表面接枝氨基（-NH_2），并通过原位合成固定DNA探针。杂交采用夹心杂交方式完成。在银增强液的放大作用下，用扫描仪记录杂交信号。靶DNA浓度低至100 fM。互补序列和错配序列被清晰区分，完美匹配、单碱基错配、2碱基错配和3碱基错配的背景减去灰度值的比值为22:16:9:4。在相同的靶DNA浓度下，原位合成系统的灵敏度比点状系统高3个数量级，前者的信号强度约为后者的2倍。
The gold-label-silver-stain method (GLSS) for DNA hybridization detection has been receiving increased interest as a colorimetric detective method, demonstrating the advantages of non-radioactivity, non-quenching effect of fluorescence and simplicity for analytical equipment. A colorimetric detection based on the GLSS method was applied to DNA arrays in situ synthesized on polypropylene (PP) slices. In this paper a simple plasma treatment was employed to graft amino (-NH_2) on the polypropylene slice surfaces, where DNA probes were immobilized via in situ synthesis. Hybridization was accomplished by a sandwich hybridization format. With the amplification of Silver Enhancer Solution, the hybridization signals were recorded with a scanner. A target DNA concentration as low as 100 fM was detected. Complementary and mismatched sequences were clearly distinguished, and the ratio of the background-subtracted gray scale values for a perfect match, single-base mismatch, 2-base mismatch and 3-base mismatch is 22:16:9:4. The sensitivity of the in situ synthesis system was 3 orders of magnitude higher than that of the spotting system, and the signals of the former were about 2-times stronger than that of the latter under the same target DNA concentration.