Array Technology Laboratory, Synthetic Biology and Precision Medicine Centre: Next Generation Health Cluster, Council for Scientific and Industrial Research, Brummeria, Pretoria 0001, South Africa.
Department of Virology, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Braamfontein, Johannesburg 2000, South Africa.
Biosensors (Basel). 2023 Sep 20;13(9):894. doi: 10.3390/bios13090894.
Currently there are diagnostic tests available for human immunodeficiency virus (HIV) and tuberculosis (TB); however, they are still diagnosed separately, which can delay treatment in cases of co-infection. Here we report on a multiplex microarray technology for the detection of HIV and TB antibodies using p24 as well as TB CFP10, ESAT6 and pstS1 antigens on epoxy-silane slides. To test this technology for antigen-antibody interactions, immobilized antigens were exposed to human sera spiked with physiological concentrations of primary antibodies, followed by secondary antibodies conjugated to a fluorescent reporter. HIV and TB antibodies were captured with no cross-reactivity observed. The sensitivity of the slides was compared to that of high-binding plates. We found that the slides were more sensitive, with the detection limit being 0.000954 µg/mL compared to 4.637 µg/mL for the plates. Furthermore, stability studies revealed that the immobilized antigens could be stored dry for at least 90 days and remained stable across all pH and temperatures assessed, with pH 7.4 and 25 °C being optimal. The data collectively suggested that the HIV/TB multiplex detection technology we developed has the potential for use to diagnose HIV and TB co-infection, and thus can be developed further for the purpose.
目前已有用于检测人类免疫缺陷病毒 (HIV) 和结核病 (TB) 的诊断测试方法；然而，这些测试方法仍然是分开进行的，这可能会导致合并感染的治疗延误。在这里，我们报告了一种使用微阵列技术同时检测 HIV 和 TB 抗体的方法，该方法使用 p24 以及 TB CFP10、ESAT6 和 pstS1 抗原作为检测物，并将其固定在环氧硅烷载玻片上。为了测试该技术在抗原-抗体相互作用方面的性能，我们将固定化抗原暴露于含有生理浓度的初始抗体的人血清中，然后再将与荧光报告基团偶联的二级抗体与之结合。结果发现 HIV 和 TB 抗体能够被特异性捕获，且没有观察到交叉反应。与高结合度平板相比，我们发现载玻片具有更高的灵敏度，其检测限为 0.000954 µg/mL，而平板的检测限为 4.637 µg/mL。此外，稳定性研究表明，固定化抗原可以在干燥状态下储存至少 90 天，并且在所有评估的 pH 值和温度下均保持稳定，其中 pH 值为 7.4 和温度为 25°C 时最为稳定。综上所述，我们开发的 HIV/TB 多重检测技术具有用于诊断 HIV 和 TB 合并感染的潜力，因此可以进一步开发用于该目的。
