Grup de Sensors i Biosensors, Departament de Química, Universitat Autònoma de Barcelona, Spain.
Laboratory of Cellular Immunology, Institute of Biotechnology and Biomedicine, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Biosens Bioelectron. 2015 Dec 15;74:974-80. doi: 10.1016/j.bios.2015.07.053. Epub 2015 Jul 26.
The counting of CD4(+) T lymphocytes is a clinical parameter used for AIDS diagnosis and follow-up. As this disease is particularly prevalent in developing countries, simple and affordable CD4 cell counting methods are urgently needed in resource-limited settings. This paper describes an electrochemical magneto-actuated biosensor for CD4 count in whole blood. The CD4(+) T lymphocytes were isolated, preconcentrated and labeled from 100 μL of whole blood by immunomagnetic separation with magnetic particles modified with antiCD3 antibodies. The captured cells were labeled with a biotinylated antiCD4 antibody, followed by the reaction with the electrochemical reporter streptavidin-peroxidase conjugate. The limit of detection for the CD4 counting magneto-actuated biosensor in whole blood was as low as 44 cells μL(-1) while the logistic range was found to be from 89 to 912 cells μL(-1), which spans the whole medical interest range for CD4 counts in AIDS patients. The electrochemical detection together with the immunomagnetic separation confers high sensitivity, resulting in a rapid, inexpensive, robust, user-friendly method for CD4 counting. This approach is a promising alternative for the costly standard flow cytometry and suitable as diagnostic tool at decentralized practitioner sites in low resource settings, especially in less developed countries.
CD4(+)T 淋巴细胞计数是用于艾滋病诊断和随访的临床参数。由于这种疾病在发展中国家尤为普遍,因此在资源有限的情况下,迫切需要简单且经济实惠的 CD4 细胞计数方法。本文介绍了一种用于全血中 CD4 计数的电化学磁激活生物传感器。通过用抗 CD3 抗体修饰的磁性颗粒进行免疫磁分离,从 100 μL 全血中分离、预浓缩和标记 CD4(+)T 淋巴细胞。捕获的细胞用生物素化的抗 CD4 抗体标记,然后与电化学报告分子链霉亲和素-过氧化物酶缀合物反应。全血中 CD4 计数磁激活生物传感器的检测限低至 44 个细胞 μL(-1),而逻辑范围为 89 到 912 个细胞 μL(-1),跨越了艾滋病患者 CD4 计数的整个医学感兴趣范围。电化学检测与免疫磁分离相结合具有高灵敏度,可实现快速、廉价、稳健、用户友好的 CD4 计数方法。这种方法是昂贵的标准流式细胞术的有前途的替代方法,适合在资源匮乏的地区分散的医疗场所作为诊断工具,尤其是在欠发达国家。