A Long-Lived Phosphorescence Amplification System Integrated with Graphene Oxide and a Stable Split G-Quadruplex Protector as an Isothermal “Off–On” Biosensor for the HBV Gene,ACS Applied Bio Materials

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A Long-Lived Phosphorescence Amplification System Integrated with Graphene Oxide and a Stable Split G-Quadruplex Protector as an Isothermal “Off–On” Biosensor for the HBV Gene
ACS Applied Bio Materials ( IF 4.6 ) Pub Date : 2020-06-11 , DOI: 10.1021/acsabm.0c00481
Chung-Nga Ko ₁ , Shasha Cheng ₂ , Chung-Hang Leung ₂ , Dik-Lung Ma ₁

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Hepatitis B is a life-threatening liver infection caused by the hepatitis B virus (HBV). Chronic HBV infection is a major global health issue, causing an estimated 750,000 of deaths annually. Despite intense interest of biomedical and clinical researchers, the lack of an accurate, selective, and simple diagnostic approach for the early stage of the disease remains a major challenge. In this work, we developed a strand displacement amplification (SDA)-assisted G-quadruplex (GQ)–graphene oxide (GO) system for the accurate quantification of the HBV gene. Iridium (III) complex 2a, a red-emitting luminophore with a large Stokes shift and long-lived phosphorescence, was identified as the most effective probe for the split GQ–GO system among the eight selected complexes in the systematic optimization campaign. The length of nucleic acid bases in between the critical positions of the split GQ hybridization was also optimized using circular dichroism (CD) and phosphorescence experiments. Important rules on the proximity and flexibility of the stable split GQ–GO system were developed for the first time. This approach was selective for the wild HBV gene over the interfering anions, cations, proteins, and the mutated HBV genes differing by only one nucleotide base. Compared to the simple GQ–GO system, the SDA-assisted GQ–GO system produced a 211% higher phosphorescence restoration signal. The ability of the SDA-assisted GQ–GO system to detect the HBV gene in human serum and sheep red blood cells demonstrates the feasibility of using this system in practical applications.

中文翻译：

与氧化石墨烯和稳定分裂 G-四链体保护器集成的长寿命磷光放大系统作为 HBV 基因的等温“关-开”生物传感器

乙型肝炎是由乙型肝炎病毒 (HBV) 引起的危及生命的肝脏感染。慢性 HBV 感染是一个主要的全球健康问题，估计每年导致 750,000 人死亡。尽管生物医学和临床研究人员非常感兴趣，但缺乏针对该疾病早期阶段的准确、选择性和简单的诊断方法仍然是一项重大挑战。在这项工作中，我们开发了一种链置换扩增 (SDA) 辅助的 G-四链体 (GQ)-氧化石墨烯 (GO) 系统，用于准确定量 HBV 基因。铱 (III) 络合物2a，一种具有大斯托克斯位移和长寿命磷光的红色发光体，被确定为系统优化活动中八种选定配合物中分裂 GQ-GO 系统的最有效探针。分裂 GQ 杂交的关键位置之间的核酸碱基长度也使用圆二色性 (CD) 和磷光实验进行了优化。首次制定了关于稳定分裂 GQ-GO 系统的接近性和灵活性的重要规则。这种方法对野生 HBV 基因的选择性高于干扰阴离子、阳离子、蛋白质和突变的 HBV 基因，它们仅相差一个核苷酸碱基。与简单的 GQ-GO 系统相比，SDA 辅助的 GQ-GO 系统产生的磷光恢复信号高出 211%。

更新日期：2020-07-20

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