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Wang Y.,Suzhou University | Wang Y.,Nanjing Medical University | Shi J.,Suzhou University | Wu Y.,Nanjing Medical University | And 5 more authors.
Tumori | Year: 2012

Background. We measured the expression of microRNA (miRNA) in non-small cell lung cancer (NSCLC) tissues using the Luminex xMAP bead-based suspension array. We discuss the feasibility of employing this method to detect miRNA in NSCLC and explore its value as a high-throughput miRNA array. Methods. We performed the methodological analysis of xMAP with oligoribonucleic acid references. We detected the expression of miR-21, miR, miR-31, miR-222, miR-145 and miR 40 NSCLC cancer tissues and adjacent normal tissues by xMAP beadbased suspension array. We selected miR-191 and miR-103 as the house-keeping genes (internal control). We also analyzed the relationship between xMAP and RT-PCR. Results. The methodological analysis parameters of xMAP are quite good. The expression of miR-21, miR, miR-31 and miR-222 was higher in NSCLC tissues than in adjacent tissues, while the expression of miR-145 and miR-126 was lower in NSCLC tissues than in adjacent tissues. The expression of miR-145 and miR-126 decreased with disease progression. The intraassay and interassay coefficients of variation were lower in xMAP than in RT-PCR. xMAP proved cheaper and more flexible in detecting multiple miRNAs of one sample. Conclusions. The Luminex xMAP bead-based suspension array for detecting miRNA has many advantages, such as allowing a smaller sample size (only 2 μL), no sample amplification, fast detection, high throughput, and flexible combination of multiple detection targets. The high throughput testing technology shows a great advantage in saving time and labor. We found that the Luminex xMAP bead-based suspension array is a good and feasible method for detecting miRNA expression with high-throughput technology. © II Pensiero Scientifico Editore downloaded by EXCERPTA MEDICA. Source

Wang Y.,Capital Medical University | Wang Y.,Beijing Key Laboratory of Translational | Fang F.,Capital Medical University | Shi C.,Capital Medical University | And 6 more authors.
Clinical Biochemistry | Year: 2012

Objective: To achieve higher tumor detection efficiency, we evaluated a multiplex assay for TM analysis based on the Luminex-100 multiplex suspension bead array. Design: The assay simultaneously determined the concentrations of nine TMs in 1114 human serum specimens (546 patients with tumors, 158 patients with non-tumor inflammatory diseases, and 410 normal controls). The nine TMs were AFP, CEA, CA125, CYFRA 21-1, CA242, f-PSA, t-PSA, NSE and free β-hCG. The multiplex suspension bead assays were compared with conventional methods used in clinical laboratories. Results: The Luminex assay has the same levels of sensitivity, specificity and accuracy in the prediction of positive tumor specimens as conventional methods. Conclusion: Multiplex suspension bead arrays have promising applications in clinical laboratories. © 2012 The Canadian Society of Clinical Chemists. Source

Wang Y.,Capital Medical University | Wang Y.,Beijing Key Laboratory for Translational Medicine in Cerebrovascular Diseases | Yu J.,Genome Technology Access Center | Ren Y.,Capital Medical University | And 10 more authors.
Clinica Chimica Acta | Year: 2013

Background: A variety of immunoassays including multiplex suspension bead array have been developed for tumor marker detections; however, these assays could be compromised in their sensitivity and specificity by well-known heterophile antibody interference and hook effect. Methods: Using Luminex® multiplex suspension bead arrays, we modified protocols with two newly-developed solutions that can identify heterophile antibody interference and AFP hook effect. Effectiveness of the two solutions was assessed in serum samples from patients. Results: Concentrations of 9 tumor markers in heterophile antibody positive samples assayed with Solution A, containing murine monoclonal antibodies and mouse serum, were significantly reduced when compared with those false high signals assayed without Solution A (all p. <. 0.01). With incorporation of Solution H (fluorescent beads linked with AFP antigen), a new strategy for identification of AFP hook effect was established, and with this strategy AFP hook effect was identified effectively in serum samples with very high levels of AFP. Conclusions: Two proprietary solutions improve the identification of heterophile antibody interference and AFP hook effect. With these solutions, multiplex suspension bead arrays provide more reliable testing results in tumor marker detection where complex clinical serum samples are used. © 2013 Elsevier B.V. Source

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