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目的:采用金标记银增强技术制备检测抗HIV病毒P24、gp41、gp36、gp120蛋白抗体的蛋白质微阵列。方法:将上述抗原片段固定在醛基化玻片上,制成了能够检测多种抗体的蛋白质微阵列,同时研究了金标记银增强技术用于蛋白质微阵列检测的方法学基础。结果:采用金银染色方法的蛋白质微阵列能够准确而灵敏地检测抗HIV不同抗原的抗体。该方法被引入应用到蛋白质微阵列检测中,可以通过光学显微镜或凭肉眼观察定性结果,仅使用普通的CCD数字照像机或CCD扫描仪就可以对信号进行采集。结论:与传统的荧光标记方法相比,金标记银增强方法的灵敏度相当,而且成本低、信号稳定、操作简便,将是前者的一种通用替代方法。
OBJECTIVE: To prepare protein microarray for detection of anti-HIV virus P24, gp41, gp36, gp120 antibody by gold-labeled silver enhancement technique. Methods: The above antigen fragments were immobilized on aldehyde-based glass slides to prepare a protein microarray capable of detecting various antibodies. Meanwhile, the methodological basis of gold-labeled silver enhancement technology for protein microarray detection was studied. Results: The protein microarray using gold-silver staining method can accurately and sensitively detect antibodies against different antigens of HIV. This method has been introduced into protein microarray detection, where qualitative results can be observed by light microscopy or by naked eye, and the signal can be collected using only a normal CCD digital camera or CCD scanner. Conclusion: Compared with the traditional fluorescence labeling method, gold labeling silver enhancement method has the same sensitivity, low cost, stable signal and simple operation, which will be a common alternative method of the former.