Rapid parallelized and quantitative analysis of five pathogenic bacteria by ITS hybridization using QCM biosensor
Rapid parallelized and quantitative analysis of five pathogenic bacteria by ITS hybridization using QCM biosensor
复制标题
使用 QCM 生物传感器通过 ITS 杂交快速并行定量分析五种病原菌
DOI：
10.1016/j.snb.2010.12.053
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发表时间：
2011-07-20
影响因子：
8.4
通讯作者：
Fu, Weiling
中科院分区：
文献类型：
--
作者：
Cai, Jin;Yao, Chunyan;Fu, Weiling
基于细菌16 S-23 SrDNA内转录间隔区（ITS）的杂交分析，研制了一种2 × 5型石英晶体微天平（QCM）DNA生物传感器阵列。设计了一对通用引物，用于ITS的PCR扩增。PCR产物用生物传感器进行分析。我们使用金纳米颗粒来放大频移信号。用该生物传感器和常规细菌培养法对50份临床标本进行检测。我们发现合成的寡核苷酸或细菌细胞的频移与对数浓度之间存在线性定量关系。合成寡核苷酸的可测量浓度范围为10（-12）至10（-8）M，细菌的可测量浓度范围为1.5 x 10（2）至1.5 x 10（8）CFU/mL。该生物传感器系统可检测到10（-12）M的合成寡核苷酸或1.5 × 10（2）CFU/mL的铜绿假单胞菌。包括PCR扩增程序在内，检测可在5 h内完成。与细菌培养法相比，该生物传感器系统的检测灵敏度和特异性分别为94.12%和90.91%。两种方法检测结果差异无统计学意义（P = 0.625 > 0.05）。该生物传感器系统为临床诊断中多种病原菌的并行定量分析提供了一种快速、灵敏的方法。(C)2010 Elsevier B. V.保留所有权利。
We developed a 2 x 5 model quartz crystal microbalance (QCM) DNA biosensor array for detection of five bacteria, which based on hybridization analysis of bacterial 16S-23S rDNA internal transcribed spacer (ITS) region. A pair of universal primers was designed for PCR amplification of the ITSs. The PCR products were analyzed by the biosensor. We used gold nanoparticles to amplify the frequency shift signals. Fifty clinical samples were detected by both the biosensor and conventional bacteria culture method. We found a linear quantitative relationship between frequency shift and logarithmic concentration of synthesized oligonucleotides or bacteria cells. The measurable concentration ranged from 10(-12) to 10(-8) M for synthesized oligonucleotides and 1.5 x 10(2) to 1.5 x 10(8) CFU/mL for bacteria. The 10(-12) M of synthesized oligonucleotides or 1.5 x 10(2) CFU/mL of Pseudomonas aeruginosa could be detected by the biosensor system. The detection could be completed within 5 h including the PCR amplification procedure. Compared with bacteria culture method, the detection sensitivity and specificity of the biosensor system were 94.12% and 90.91%, respectively. There was no significant difference between these two methods (P = 0.625 > 0.05). The biosensor system provides a rapid and sensitive method for parallelized and quantitative analysis of multiple pathogenic bacteria in clinical diagnosis. (C) 2010 Elsevier B.V. All rights reserved.