Development and Validation of Multiplex Liquid Bead Array Assay for the Simultaneous Expression of 14 Genes in Circulating Tumor Cells

被引:4
|
作者
Parisi, Cleo [1 ]
Markou, Athina [1 ]
Strati, Areti [1 ]
Kasimir-Bauer, Sabine [2 ]
Lianidou, Evi S. [1 ]
机构
[1] Univ Athens, Dept Chem, Lab Analyt Chem, Anal Circulating Tumor Cells Lab, GR-15771 Athens, Greece
[2] Univ Duisburg Essen, Univ Hosp Essen, Dept Gynecol & Obstet, D-45122 Essen, Germany
关键词
RNA-POSITIVE CELLS; BREAST-CANCER; PERIPHERAL-BLOOD; PROGNOSTIC VALUE; MOLECULAR CHARACTERIZATION; ADJUVANT CHEMOTHERAPY; CYTOKERATIN-19; CHALLENGES; FUTURE; HER2;
D O I
10.1021/acs.analchem.8b04975
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Liquid biopsy, based on the molecular information extracted from circulating tumor cells (CTCs) and circulating tumor DNA (ctDNA), offers the possibility to characterize the evolution of a solid tumor in real time and is highly important for diagnostic and therapeutic purposes. The aim of the present study was the development and validation of a novel liquid bead array methodology for the molecular characterization of CTCs and its application in breast cancer. In the present study we developed and evaluated a multiplex polymerase chain reaction (PCR)-coupled liquid bead array (MLBA) assay for studying simultaneously the expression of 14 genes in CTCs. The 14-gene MLBA assay is characterized by high analytical specificity, sensitivity, and reproducibility. The analytical performance of the 14-gene MLBA assay was compared with a commercially available test (AdnaTest Breast Cancer, Qiagen, Germany) and our previously described multiplex quantitative reverse transcription PCR (RT-qPCR) assays. The developed assay has the potential to be further expanded in order to include up to 100 gene targets. The assay is highly specific for each target gene and is not affected by the numerous primers and probes used for multiplexing; hence, it constitutes a sample-, cost-, and time-saving analysis.
引用
收藏
页码:3443 / 3451
页数:9
相关论文
共 50 条
  • [41] Development and validation of a multiplex PCR assay for simultaneous detection of chicken, turkey and duck in processed meat products
    Kim, Mi-Ju
    Yoo, Insuk
    Yang, Seung-Min
    Suh, Seung-Man
    Kim, Hae-Yeong
    INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY, 2018, 53 (12): : 2673 - 2679
  • [42] Development of a Multiplex Bead Assay for Simultaneous Serodiagnosis of Antibodies against Mycobacterium bovis, Brucella suis, and Trichinella spiralis in Wild Boar
    Touloudi, Antonia
    Valiakos, George
    Cawthraw, Shaun
    Kostoulas, Polychronis
    Gortazar, Christian
    Boadella, Mariana
    Giannakopoulos, Alexios
    Birtsas, Periklis
    Sofia, Marina
    Athanasiou, Labrini V.
    Satra, Maria
    Athanasakopoulou, Zoi
    Kantere, Maria
    Spyrou, Vassiliki
    Petrovska, Liljana
    Billinis, Charalambos
    MICROORGANISMS, 2021, 9 (05)
  • [43] Development and Validation of a Quantitative, Multiplex Gene Expression Assay for Components of the Endocrine Growth Axis in Salmon
    Luckenbach, J. A.
    Metzger, D. C.
    Dickey, J. T.
    Swanson, P.
    Beckman, B. R.
    INTEGRATIVE AND COMPARATIVE BIOLOGY, 2010, 50 : E261 - E261
  • [44] Development and Validation of a Multiplex, Bead-based Assay to Detect Antibodies Directed Against SARS-CoV-2 Proteins
    Bray, Robert A.
    Lee, Jar-How
    Brescia, Peter
    Kumar, Deepali
    Nong, Thoa
    Shih, Remi
    Woodle, E. Steve
    Maltzman, Jonathan S.
    Gebel, Howard M.
    TRANSPLANTATION, 2021, 105 (01) : 79 - 89
  • [45] Development of a Molecular Multimarker Assay for the Analysis of Circulating Tumor Cells in Adenocarcinoma Patients
    de Albuquerque, Andreia
    Kubisch, Ilja
    Ernst, Dominikus
    Breier, Georg
    Stamminger, Gudrun
    Fersis, Nikos
    Stoelzel, Ulrich
    Boese-Landgraf, Joachim
    Eichler, Astrid
    Kaul, Sepp
    CLINICAL LABORATORY, 2012, 58 (5-6) : 373 - 384
  • [46] Clinical assay development and validation of multiplex immunofluorescent (mIHC) marker panel for evaluation of tumor infiltration myeloid cells in FFPE tissue sections
    Yi, Lan
    Juco, Jonathan
    Mahmood, Ashhad
    Laterza, Omar
    Garrido, Charo
    Yi, Lan
    JOURNAL FOR IMMUNOTHERAPY OF CANCER, 2019, 7
  • [47] Development and Validation of an Efficient Multiplex PCR Assay for Simultaneous Detection of Six Common Foodborne Pathogens and Hygiene Indicators
    Ngamwongsatit, Natharin
    Chaturongakul, Soraya
    Aunpad, Ratchaneewan
    FOODBORNE PATHOGENS AND DISEASE, 2023, 20 (06) : 222 - 229
  • [48] Development and initial clinical testing of a multiplex assay evaluating histone modifier p300 acetylation in prostate cancer circulating tumor cells
    Jarrard, David Frazier
    Filon, Mikolaj
    Yang, Bing
    Schehr, Jennifer L.
    Singh, Anupama
    Bigarella, Marcelo
    Denu, John M.
    Lang, Joshua Michael
    JOURNAL OF CLINICAL ONCOLOGY, 2023, 41 (16)
  • [49] Multiplex protein detection on circulating tumor cells from liquid biopsies using imaging mass cytometry
    Gerdtsson, Erik
    Pore, Milind
    Thiele, Jana-Aletta
    Gerdtsson, Anna Sandstrom
    Malihi, Paymaneh D.
    Nevarez, Rafael
    Kolatkar, Anand
    Velasco, Carmen Ruiz
    Wix, Sophia
    Singh, Mohan
    Carlsson, Anders
    Zurita, Amado J.
    Logothetis, Christopher
    Merchant, Akil A.
    Hicks, James
    Kuhn, Peter
    CONVERGENT SCIENCE PHYSICAL ONCOLOGY, 2018, 4 (01):
  • [50] Crohn's Colitis: Development of a multiplex gene expression assay comparing mRNA levels of susceptibility genes
    Russo, Edda
    Lombardelli, Letizia
    Giudici, Francesco
    Cavalli, Tiziana
    Ficari, Ferdinando
    Fazi, Marilena
    Scaringi, Stefano
    Biancone, Livia
    Logiodice, Federica
    Nesi, Mariateresa
    Latiano, Anna
    Annese, Vito
    Torcia, Maria Gabriella
    Bechi, Paolo
    Tonelli, Francesco
    Piccinni, Marie-Pierre
    Malentacchi, Cecilia
    CLINICS AND RESEARCH IN HEPATOLOGY AND GASTROENTEROLOGY, 2017, 41 (04) : 435 - 444