Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education), Bio-X Institutes, Shanghai Jiao Tong University, Shanghai, 200030, China.
Division of Physical Biology & Bioimaging Center, Shanghai Synchrotron Radiation Facility, CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai, 201800, China.
Small. 2018 Feb;14(6). doi: 10.1002/smll.201701718. Epub 2017 Dec 28.
The hepatitis B virus (HBV) genotyping may profoundly affect the accurate diagnosis and antiviral treatment of viral hepatitis. Existing genotyping methods such as serological, immunological, or molecular testing are still suffered from substandard specificity and low sensitivity in laboratory or clinical application. In a previous study, a set of high-efficiency hybridizable DNA origami-based shape ID probes to target the templates through which genetic variation could be determined in an ultrahigh resolution of atomic force microscopy (AFM) nanomechanical imaging are established. Here, as a further confirmatory research to explore the sensitivity and applicability of this assay, differentially predesigned DNA origami shape ID probes are also developed for precisely HBV genotyping. Through the specific identification of visualized DNA origami nanostructure with clinical HBV DNA samples, the genetic variation information of genotypes can be directly identified under AFM. As a proof-of-concept, five genotype B and six genotype C are detected in 11 HBV-infected patients' blood DNA samples of Han Chinese population in the single-blinded test. The AFM image-based DNA origami shape ID genotyping approach shows high specificity and sensitivity, which could be promising for virus infection diagnosis and precision medicine in the future.
乙型肝炎病毒(HBV)基因分型可能会深刻影响病毒性肝炎的准确诊断和抗病毒治疗。现有的基因分型方法,如血清学、免疫学或分子检测,在实验室或临床应用中仍然存在特异性差、灵敏度低的问题。在之前的一项研究中,建立了一套高效杂交 DNA 折纸基形状 ID 探针,通过原子力显微镜(AFM)纳米力学成像的超高分辨率,可以针对模板确定遗传变异。在这里,作为进一步探索该检测方法的灵敏度和适用性的确认性研究,还开发了针对 HBV 基因分型的差异化预设计 DNA 折纸形状 ID 探针。通过与临床 HBV DNA 样本进行可视化 DNA 折纸纳米结构的特异性识别,可以直接在 AFM 下识别基因型的遗传变异信息。作为概念验证,在 11 例汉族乙型肝炎病毒感染者的血液 DNA 样本中,用该方法检测到了 5 种基因型 B 和 6 种基因型 C。基于 AFM 图像的 DNA 折纸形状 ID 基因分型方法具有很高的特异性和灵敏度,有望在未来用于病毒感染诊断和精准医疗。