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  • br Please cite this article as Park YR et

    2020-03-24


    Please cite this article as: Park YR et al., Absence of Pifithrin-α (PFTα) receptor is associated with worse oncologic outcome in patients who were received neoadjuvant chemotherapy for breast cancer, Asian Journal of Surgery, https://doi.org/10.1016/j.asjsur.2019.05.010 Biosensors and Bioelectronics 142 (2019) 111523
    Contents lists available at ScienceDirect
    Biosensors and Bioelectronics
    journal homepage: www.elsevier.com/locate/bios
    Absolute quantification and analysis of extracellular vesicle lncRNAs from T the peripheral blood of patients with lung cancer based on multi-colour fluorescence chip-based digital PCR
    Yanan Baia,b,1, Youlan Qua,c,1, Zhenhua Wua, Yijiu Rend, Zule Chenga,b, Yunxing Lua,b, Jie Hue, Jiatao Louf, Jianlong Zhaoa, Chang Chend,∗∗, Hongju Maoa,b,∗ a State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China
    b Center of Materials Science and Optoelectronics Engineering, University of Chinese Academy of Sciences, Beijing, 100049, China
    c School of Stomatology, Dalian Medical University, Dalian, 116044, China
    d Department of Thoracic Surgery, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, 200443, China
    e Department of Pulmonary Medicine, Zhongshan Hospital, Fudan University, Shanghai, 200032, China
    f Department of Laboratory Medicine, Shanghai Chest Hospital, Shanghai Jiaotong University, Shanghai, 200030, China
    Keywords:
    Lung cancer
    Extracellular vesicles
    lncRNA
    Early diagnosis Multi-colour fluorescence 
    Emerging evidence indicates that extracellular vesicle (EV) long non-coding ribonucleic acids (lncRNAs) in lung cancer may be clinically useful biomarkers for early diagnosis using liquid biopsy. However, the extremely low quantities of EV-lncRNAs in peripheral blood are a major challenge for multi-target detection. In this study, we developed a new multi-colour fluorescence digital PCR EV-lncRNA (miDER) analysis chip, and then demon-strated its ability to quickly and accurately analyse the levels of two target genes and one reference gene from peripheral blood. Under the miDER assay, the limit of detection of the target gene from peripheral blood was 10 copies/μL. Based on multiplex assay, the expression levels of two lung cancer-related genes (SLC9A3-AS1 and PCAT6) in patients with lung cancer (n = 32) and healthy controls (n = 30) showed a significant difference between the two groups (P < 0.001; two-tailed t-test). A receiver operating characteristic (ROC) curve analysis was used to evaluate the discrimination ability of these lncRNAs. The combination of two lncRNAs in the miDER assay yielded a higher area under curve (AUC) value of 0.811 (95% CI = 0.705–0.918). Moreover, to determine the absolute quantitation capacity of the miDER assay, we compared the results to those obtained by quantitative real-time polymerase chain reaction (qPCR), demonstrating that the miDER assay is more sensitive than qPCR. The multiplex assay based on the miDER could provide a new solution for the multi-index combined detection of trace EV-lncRNAs in body fluids and demonstrate the use of EV-lncRNAs as biomarkers for lung tumour biopsy.
    1. Introduction
    Lung cancer is the most common malignant tumour with the highest incidence worldwide, accounting for the largest number of cancer deaths. Due to the complexity of lung tissues, the absence of obvious symptoms in the early stage, and the easy spread and metastasis, lung cancer is typically diagnosed at an intermediate or advanced stage. The early detection of lung cancer is mainly based on imaging and tumour protein markers (Hirsch et al., 2001; Wulfkuhle et al., 2003). Advanced imaging techniques, such as multi-slice spiral computed tomography