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Simultaneous multi-analyte urinary protein assay for bladder cancer detection.

Rosser CJ, Dai Y, Miyake M, Zhang G, Goodison S - BMC Biotechnol. (2014)

Bottom Line: The following analytes, interleukin 8 (IL8), matrix metallopeptidase 9 (MMP9), and vascular endothelial growth factor A (VEGFA) were monitored using Q-plex, a customized multiplex ELISA system from Quansys Biosciences, and individual target commercial ELISA kits.The Q-plex assay achieved an overall diagnostic sensitivity of 0.93 and specificity of 0.81, and the individual target ELISA assays achieved an overall sensitivity of 0.77 and specificity of 0.91.Based on these encouraging preliminary data, we believe that the Q-Plex technology is a viable platform that can be exploited as an efficient, highly accurate tool to quantitate multiplex panels of diagnostic proteins in biologic specimens.

View Article: PubMed Central - HTML - PubMed

Affiliation: MD Anderson Cancer Center Orlando, Orlando, FL, USA. crosser@cc.hawaii.edu.

ABSTRACT

Background: The ability to accurately measure multiple proteins simultaneously in a single assay has the potential to markedly improve the efficiency of a myriad of clinical assays. Here, we tested the performance of a new, multiplex protein array platform to quantitate three bladder cancer-associated proteins in urine samples. The following analytes, interleukin 8 (IL8), matrix metallopeptidase 9 (MMP9), and vascular endothelial growth factor A (VEGFA) were monitored using Q-plex, a customized multiplex ELISA system from Quansys Biosciences, and individual target commercial ELISA kits. The performance of the two approaches was compared by evaluating the diagnostic accuracy of the biomarker assays in samples from a cohort of 73 subjects of known bladder cancer status.

Results: The combination biomarker panel analyses revealed an AUROC value of 0.9476 for the Q-plex assay, and 0.9119 for the combination of the single-target ELISA assays. The Q-plex assay achieved an overall diagnostic sensitivity of 0.93 and specificity of 0.81, and the individual target ELISA assays achieved an overall sensitivity of 0.77 and specificity of 0.91.

Conclusion: Based on these encouraging preliminary data, we believe that the Q-Plex technology is a viable platform that can be exploited as an efficient, highly accurate tool to quantitate multiplex panels of diagnostic proteins in biologic specimens.

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Related in: MedlinePlus

Diagnostic performance of bladder cancer-associated molecular panel comprised of three biomarkers. ROC curves were plotted to illustrate the performance characteristics of the 3-biomarker signature for the detection of bladder cancer in urine samples using the Q-Plex multiplex assay (solid line) and commercial ELISA assays (dotted line). Based on the area under the ROC curve (AUROC), Youden Index cutoff values that maximized the sum of sensitivity and specificity were determined for the combination of biomarkers. The Q-Plex multiplex assay achieved an overall sensitivity of 0.93 and specificity of 0.81 (AUROC 0.9476). The combination of data from the individual target ELISA assays achieved an overall sensitivity of 0.77 and specificity of 0.91 (AUROC 0.9119). Thus, the comparison of multiplex results with standard ELISA of these three diagnostic biomarkers showed similar results and trends.
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Figure 1: Diagnostic performance of bladder cancer-associated molecular panel comprised of three biomarkers. ROC curves were plotted to illustrate the performance characteristics of the 3-biomarker signature for the detection of bladder cancer in urine samples using the Q-Plex multiplex assay (solid line) and commercial ELISA assays (dotted line). Based on the area under the ROC curve (AUROC), Youden Index cutoff values that maximized the sum of sensitivity and specificity were determined for the combination of biomarkers. The Q-Plex multiplex assay achieved an overall sensitivity of 0.93 and specificity of 0.81 (AUROC 0.9476). The combination of data from the individual target ELISA assays achieved an overall sensitivity of 0.77 and specificity of 0.91 (AUROC 0.9119). Thus, the comparison of multiplex results with standard ELISA of these three diagnostic biomarkers showed similar results and trends.

Mentions: After confirming the robustness of the multiplex assay for these specific biomarkers, it was used to test a voided urine sample set. Seventy-three clinical urine samples from MD Anderson Cancer Center Orlando were made available for analysis. Demographics and disease characteristics of the entire cohort are summarized in Table 1. The ability of each of the test biomarkers within the multiplex array to predict the presence of bladder cancer was analyzed using nonparametric ROC analyses, according to National Cancer Institute guidelines [20]. Urinary IL8 was the most accurate single biomarker for bladder cancer detection with an AUROC of 0.907 (95% CI: 0.830 - 0.985), a sensitivity of 90%, specificity of 86%, PPV of 82% and NPV of 92%. Urinary MMP9 data generated an AUROC of 0.533 (95% CI: 0.392 - 0.674), sensitivity of 45%, specificity of 76%, PPV of 58% and NPV of 65%, and VEGFA as an individual analyte was noted to have an AUROC of 0.524 (95% CI: 0.386 - 0.661), sensitivity of 17%, specificity of 95%, PPV of 71% and NPV of 61%. Through combinatorial analysis of all 3 biomarkers using optimal cutoff values defined by Youden index calculations, the AUROC (Figure 1) for the diagnostic panel using the multiplex array was 0.9476 [95% CI: 0.903 - 0.992]. The combination assay achieved an overall sensitivity of 0.93, specificity of 0.81, PPV of 0.78 and NPV of 0.94 for bladder cancer classification.


Simultaneous multi-analyte urinary protein assay for bladder cancer detection.

Rosser CJ, Dai Y, Miyake M, Zhang G, Goodison S - BMC Biotechnol. (2014)

Diagnostic performance of bladder cancer-associated molecular panel comprised of three biomarkers. ROC curves were plotted to illustrate the performance characteristics of the 3-biomarker signature for the detection of bladder cancer in urine samples using the Q-Plex multiplex assay (solid line) and commercial ELISA assays (dotted line). Based on the area under the ROC curve (AUROC), Youden Index cutoff values that maximized the sum of sensitivity and specificity were determined for the combination of biomarkers. The Q-Plex multiplex assay achieved an overall sensitivity of 0.93 and specificity of 0.81 (AUROC 0.9476). The combination of data from the individual target ELISA assays achieved an overall sensitivity of 0.77 and specificity of 0.91 (AUROC 0.9119). Thus, the comparison of multiplex results with standard ELISA of these three diagnostic biomarkers showed similar results and trends.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4230247&req=5

Figure 1: Diagnostic performance of bladder cancer-associated molecular panel comprised of three biomarkers. ROC curves were plotted to illustrate the performance characteristics of the 3-biomarker signature for the detection of bladder cancer in urine samples using the Q-Plex multiplex assay (solid line) and commercial ELISA assays (dotted line). Based on the area under the ROC curve (AUROC), Youden Index cutoff values that maximized the sum of sensitivity and specificity were determined for the combination of biomarkers. The Q-Plex multiplex assay achieved an overall sensitivity of 0.93 and specificity of 0.81 (AUROC 0.9476). The combination of data from the individual target ELISA assays achieved an overall sensitivity of 0.77 and specificity of 0.91 (AUROC 0.9119). Thus, the comparison of multiplex results with standard ELISA of these three diagnostic biomarkers showed similar results and trends.
Mentions: After confirming the robustness of the multiplex assay for these specific biomarkers, it was used to test a voided urine sample set. Seventy-three clinical urine samples from MD Anderson Cancer Center Orlando were made available for analysis. Demographics and disease characteristics of the entire cohort are summarized in Table 1. The ability of each of the test biomarkers within the multiplex array to predict the presence of bladder cancer was analyzed using nonparametric ROC analyses, according to National Cancer Institute guidelines [20]. Urinary IL8 was the most accurate single biomarker for bladder cancer detection with an AUROC of 0.907 (95% CI: 0.830 - 0.985), a sensitivity of 90%, specificity of 86%, PPV of 82% and NPV of 92%. Urinary MMP9 data generated an AUROC of 0.533 (95% CI: 0.392 - 0.674), sensitivity of 45%, specificity of 76%, PPV of 58% and NPV of 65%, and VEGFA as an individual analyte was noted to have an AUROC of 0.524 (95% CI: 0.386 - 0.661), sensitivity of 17%, specificity of 95%, PPV of 71% and NPV of 61%. Through combinatorial analysis of all 3 biomarkers using optimal cutoff values defined by Youden index calculations, the AUROC (Figure 1) for the diagnostic panel using the multiplex array was 0.9476 [95% CI: 0.903 - 0.992]. The combination assay achieved an overall sensitivity of 0.93, specificity of 0.81, PPV of 0.78 and NPV of 0.94 for bladder cancer classification.

Bottom Line: The following analytes, interleukin 8 (IL8), matrix metallopeptidase 9 (MMP9), and vascular endothelial growth factor A (VEGFA) were monitored using Q-plex, a customized multiplex ELISA system from Quansys Biosciences, and individual target commercial ELISA kits.The Q-plex assay achieved an overall diagnostic sensitivity of 0.93 and specificity of 0.81, and the individual target ELISA assays achieved an overall sensitivity of 0.77 and specificity of 0.91.Based on these encouraging preliminary data, we believe that the Q-Plex technology is a viable platform that can be exploited as an efficient, highly accurate tool to quantitate multiplex panels of diagnostic proteins in biologic specimens.

View Article: PubMed Central - HTML - PubMed

Affiliation: MD Anderson Cancer Center Orlando, Orlando, FL, USA. crosser@cc.hawaii.edu.

ABSTRACT

Background: The ability to accurately measure multiple proteins simultaneously in a single assay has the potential to markedly improve the efficiency of a myriad of clinical assays. Here, we tested the performance of a new, multiplex protein array platform to quantitate three bladder cancer-associated proteins in urine samples. The following analytes, interleukin 8 (IL8), matrix metallopeptidase 9 (MMP9), and vascular endothelial growth factor A (VEGFA) were monitored using Q-plex, a customized multiplex ELISA system from Quansys Biosciences, and individual target commercial ELISA kits. The performance of the two approaches was compared by evaluating the diagnostic accuracy of the biomarker assays in samples from a cohort of 73 subjects of known bladder cancer status.

Results: The combination biomarker panel analyses revealed an AUROC value of 0.9476 for the Q-plex assay, and 0.9119 for the combination of the single-target ELISA assays. The Q-plex assay achieved an overall diagnostic sensitivity of 0.93 and specificity of 0.81, and the individual target ELISA assays achieved an overall sensitivity of 0.77 and specificity of 0.91.

Conclusion: Based on these encouraging preliminary data, we believe that the Q-Plex technology is a viable platform that can be exploited as an efficient, highly accurate tool to quantitate multiplex panels of diagnostic proteins in biologic specimens.

Show MeSH
Related in: MedlinePlus