Department of Chemical & Biomolecular Engineering (BK 21 program), KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Biosens Bioelectron. 2011 Jul 15;26(11):4314-9. doi: 10.1016/j.bios.2011.04.024. Epub 2011 May 30.
The results of an investigation aimed at the development of a DNA chip for the detection of genitourinary infections are described. Through analysis of over 35,000 clinical cases, 14 pathogens which are most abundantly found among Koreans were selected and candidate sequences for capture probes were accordingly chosen by considering their sequences and β-globin house-keeping gene. Among this group, the most suitable capture probe sequences were selected by employing repeated chip tests in which they are immobilized on a glass chip by using a recently developed novel gold nanoparticles-based method. A multiplex PCR method was established to generate fluorescence-labeled sequences for all 14 pathogens along with the β-globin gene. By using optimized hybridization conditions, the final chip was constructed and employed to diagnose reliably both single and multiple infections in clinical human samples for 14 target pathogens. The results show that the novel chip methodology serves as a highly reliable and convenient tool for the diagnosis of Sexually Transmitted Diseases (STDs). Furthermore, this study has its great significance in that it demonstrates the entire process from statistical analysis of a large number of clinical cases to the final development of STD DNA chip just ready to be applied or commercialized in the clinical diagnostic field.
本文描述了一种用于检测泌尿生殖系统感染的 DNA 芯片的开发研究结果。通过对超过 35000 例临床病例的分析，选择了在韩国最常见的 14 种病原体，并考虑到它们的序列和β-珠蛋白管家基因选择了捕获探针的候选序列。在这组中，通过使用最近开发的基于新型金纳米粒子的方法将它们固定在玻璃芯片上的重复芯片测试中选择了最合适的捕获探针序列。建立了一种多重 PCR 方法，用于生成所有 14 种病原体和β-珠蛋白基因的荧光标记序列。通过优化杂交条件，构建了最终的芯片，并用于可靠地诊断临床人类样本中的 14 种目标病原体的单一和多种感染。结果表明，新型芯片方法学是一种高度可靠和方便的性传播疾病（STDs）诊断工具。此外，这项研究具有重要意义，因为它展示了从大量临床病例的统计分析到 STD DNA 芯片最终开发的整个过程，该芯片已经准备好在临床诊断领域应用或商业化。
