National Center for International Research of Bio-targeting Theranostics, Guangxi Key Laboratory of Bio-targeting Theranostics, Collaborative Innovation Center for Targeting Tumor Diagnosis and Therapy, Guangxi Medical University, Nanning, Guangxi 530021, China.
Analyst. 2019 Jun 21;144(12):3843-3852. doi: 10.1039/c8an02342e. Epub 2019 May 17.
Acute leukemia is a malignant clonal disease of hematopoietic stem cells with a high prevalence and mortality rate. However, there are no efficient tools to facilitate early diagnosis and treatment of leukemia. Therefore, development of new methods for the early diagnosis and prevention of leukemia, especially non-invasive diagnosis at the cellular level, is imperative. Here, a label-free signal-on fluorescence aptasensor based on terbium(iii)-aptamer (Tb-apt) was applied for the detection of leukemia. The aptamer sensitizes the fluorescence of Tb and forms the strong fluorescent Tb-apt probe. The target cells, the T-cell acute lymphoblastic leukemia cell line (CCRF-CEM) combined with the Tb-apt probe to form the Tb-apt-CEM complex, were removed by centrifugation, and the supernatant containing a small amount of the Tb-apt probe was detected using a fluorescence spectrophotometer. The logarithm of cell concentration showed a good linear relationship (R = 0.9881) with the fluorescence signal. The linear range for CCRF-CEM detection was 5-5 × 10 cells per ml, while the detection limit was 5 cells per ml of the binding buffer. Clinical samples were collected from 100 cases, and the specificity and positive rates detected by this method were up to 94% and 90%, respectively. Therefore, a single-stranded DNA-sensitized terbium(iii) luminescence method diagnostic was developed which is rapid, sensitive, and economical and can be used for diagnosis of various types of leukemia at the early stage.
急性白血病是一种造血干细胞恶性克隆性疾病，具有较高的发病率和死亡率。然而，目前尚无有效的工具来促进白血病的早期诊断和治疗。因此，迫切需要开发新的方法来早期诊断和预防白血病，特别是在细胞水平上进行非侵入性诊断。在这里，我们应用了一种基于铽（III）-适配体（Tb-apt）的无标记信号开启荧光适体传感器来检测白血病。该适配体能敏化铽的荧光，并形成强荧光的 Tb-apt 探针。将靶细胞（与 Tb-apt 探针结合的 T 细胞急性淋巴细胞白血病细胞系（CCRF-CEM））通过离心去除，并用荧光分光光度计检测含有少量 Tb-apt 探针的上清液。细胞浓度的对数与荧光信号呈良好的线性关系（R = 0.9881）。CCRF-CEM 的检测线性范围为 5-5×10 个细胞/ml，检测限为结合缓冲液中 5 个细胞/ml。从 100 例临床样本中收集到了该方法的特异性和阳性率分别高达 94%和 90%。因此，开发了一种基于单链 DNA 敏化的铽（III）发光方法诊断试剂盒，该试剂盒具有快速、灵敏、经济的特点，可用于早期诊断各种类型的白血病。
