Scale-Up Evaluation of a Composite Tumor Marker Assay for the Early Detection of Renal Cell Carcinoma
<p>The serum concentration of NNMT, LCP1, and NM23A was measured with the 3-plex assay. The median concentration of NNMT was 30.1-fold higher in patients (2830 pg/mL) with RCC than in controls (94 pg/mL) (<b>A</b>); the median concentration of LCP1 was 3.4-fold higher in RCC patients (11,800 pg/mL in controls, 40,232 pg/mL in patients) (<b>B</b>); and the median concentration of NM23A was 2.4-fold higher in RCC patients (429 pg/mL in controls, 1016 pg/mL in patients) (<b>C</b>).</p> "> Figure 2
<p>Comparison of Receiver Operating Characteristic (ROC) curve for NNMT, LCP1 and NM23A and the combination assay: (<b>A</b>) ROC curve for each of the three markers and scores from the composite assay and (<b>B</b>) dot plot of ROC for score from the composite assay. NNMT, Nicotinamide N-methyltransferase; LCP1, L-plastin; NM23A, Nonetastatic cells 1 protein; Sens, Sensitivity; Spec, Specificity.</p> "> Figure 3
<p>Distribution of serum concentration of NNMT (<b>A</b>), LCP1 (<b>B</b>), and NM23A (<b>C</b>) measured with the 3-plex assay. A box-and-whisker plot was used to represent the distribution of the three markers. The central box represents the values from the lower to upper quartile (25th to 75th percentile). The middle line represents the median. NNMT, Nicotinamide N-methyltransferase; LCP1, L-plastin; NM23A, Nonetastatic cells 1 protein.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Clinical Samples
2.2. Sample Preparation
2.3. Recombinant Antigens and Antibodies
2.4. Antibody Conjugation to Beads
2.5. Multiplexed Microsphere Bead-Based Immunoassay
2.6. Analytical Performance Validation
2.7. Data Analysis
3. Results
3.1. Validation of Analytical Performance
3.2. Differential Serum Levels of NNMT, LCP1 and NM23A in Control Individuals and RCC Patients
3.3. Composite Markers (NNMT, LCP1 and NM23A Together) Showed Improved Clinical Performance
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Combined Total (n = 1042) | Test Group (n = 1042) | ||||||||
---|---|---|---|---|---|---|---|---|---|
Control (n = 500) | Control (n = 500) | ||||||||
Healthy (n = 500) | 500 (100%) | ||||||||
Benign (n = 0) | 0 (0%) | ||||||||
Sex | Male | Female | Age Median | ||||||
188 (38%) | 312 (62%) | 46.8 | |||||||
RCC (n = 542) | RCC (n = 542) | ||||||||
Sex | Male | Female | Age Median | ||||||
352 (65%) | 190 (35%) | 60 | |||||||
Cell Type | Pathologic T(pT) Stage | ||||||||
I | II | III | IV | Missing | |||||
Clear Cell RCC (n = 351) | 351 (64.8%) | 268 | 39 | 40 | 3 | 1 | |||
Papillary RCC (n = 96) | 96 (17.7%) | 83 | 6 | 6 | 1 | 0 | |||
Chromophobe RCC (n = 71) | 71 (13.1%) | 55 | 10 | 6 | 0 | 0 | |||
Unclassified/Other RCC (n = 25) | 25 (4.6%) | 15 | 3 | 2 | 0 | 5 | |||
Total | 421(77.5%) | 58(10.7%) | 54(9.9%) | 4(0.7%) | 6(1.1%) |
Markers | NNMT | LCP1 | NM23A | NNMT, LCP1, NM23A Score |
Cut-point for scoring (pg/mL) | >147 | >17,800 | >520 | >1 |
Combined Total Group (n = 1042) | ||||
Median Concentration (pg/mL) (range, lower and upper 95% CI) | ||||
Control group (n = 500) | 94.0 (89.6–99.0) | 11,800.5 (11,566.11–11,984.2) | 429.0 (419.5–439.0) | |
Kidney cancer group (n = 542) | 2830 (2361.3–3886.2) | 40,232.5 (37,706.7–40,232.0) | 1016.5 (753.1–1340.4) | |
Mann–Whitney test | p < 0.0001 | p < 0.0001 | p < 0.0001 | |
AUC (95% CI)) | 0.833 (0.809–0.855) | 0.844 (0.820–0.865) | 0.601 (0.570–0.631) | 0.917 (0.898–0.933) |
Sensitivity (%) (95% CI) | 90.2 (87.4–92.6) | 82.5 (79.0–85.6) | 56.2 (52.0–60.5) | 87.1 (84.0–89.8) |
Specificity (%) (95% CI) | 77.1 (73.4–80.6) | 86.8 (83.5–89.6) | 85.8 (82.4–88.7) | 89.4 (86.4–92.0) |
+PV (%) (95% CI) | 89.5 (86.4–92.1) | 87.1 (83.9–89.9) | 81.1 (76.8–84.9) | 87.2 (80.0–92.5) |
−PV (%) (95% CI) | 78.4 (74.8–81.7) | 82.0 (78.5–85.2) | 64.4 (60.6–68.1) | 89.9 (87.0–92.3) |
Specificity = 90% Sensitivity (%) (95% CI) | 77.4 (73.7–80.9) | 67.3 (63.2–71.3) | 41.3 (41.1–49.7) | 87.0 (86.4–92.0) |
Sensitivity % | |||
---|---|---|---|
Cell Type | |||
Marker | Clear Cell Carcinoma | Papillary Carcinoma | Chromophobe Carcinoma |
NNMT at cut-off > 147 pg/mL | 81 (285/351) | 84 (81/96) | 85 (60/71) |
LCP1 at cut-off > 17,800 pg/mL | 78 (274/351) | 89 (85/96) | 83 (59/71) |
NM23A at cut-off > 520 pg/mL | 56 (196/351) | 57 (55/96) | 45 (32/71) |
NNMT + LCP1 + NM23A score at cut-off > 1 | 86 (302/351) | 88 (84/96) | 86 (61/71) |
Specificity (%) | Sensitivity (%) | ||||
---|---|---|---|---|---|
Control | Pathologic T(pT) stage | ||||
Stage I | Stage II | Stage III | Stage IV | ||
NNMT at cut-off > 147 pg/mL | 83.0 (415/500) | 83 (312/375) | 78 (64/82) | 82 (32/39) | 85 (33/39) |
NNMT, LCP1, NM23A score at cut-off > 1 | 89.4 (447/500) | 87 (326/375) | 89 (73/82) | 87 (34/39) | 85 (33/39) |
Control Samples (n = 500) | Patient Samples (n = 542) | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
Percentage of population by age | Percentage of population by age | |||||||||||
20s | 30s | 40s | 50s | 60s | 70s | 20s | 30s | 40s | 50s | 60s | 70s | 80s |
15% | 12% | 28% | 26% | 17% | 0.6% | 2% | 4% | 10% | 25% | 38% | 17% | 3% |
Specificity (%) | Sensitivity (%) | |||||||||||
92.3% | 90.5% | 86.5% | 90.0% | 89.7% | 100% | 75% | 86% | 74% | 85% | 92% | 91% | 100% |
Percentage of population by sex | Specificity | Percentage of population by sex | Sensitivity | |||||||||
Male | Female | Male | Female | Male | Female | Male | Female | |||||
38% | 62% | 91.7% | 88.1% | 65% | 35% | 88.6% | 88.0% |
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Kim, D.S.; Ham, W.S.; Jang, W.S.; Cho, K.S.; Choi, Y.D.; Kang, S.; Kim, B.; Kim, K.J.; Lim, E.J.; Rha, S.Y.; et al. Scale-Up Evaluation of a Composite Tumor Marker Assay for the Early Detection of Renal Cell Carcinoma. Diagnostics 2020, 10, 750. https://doi.org/10.3390/diagnostics10100750
Kim DS, Ham WS, Jang WS, Cho KS, Choi YD, Kang S, Kim B, Kim KJ, Lim EJ, Rha SY, et al. Scale-Up Evaluation of a Composite Tumor Marker Assay for the Early Detection of Renal Cell Carcinoma. Diagnostics. 2020; 10(10):750. https://doi.org/10.3390/diagnostics10100750
Chicago/Turabian StyleKim, Dong Su, Won Sik Ham, Won Sik Jang, Kang Su Cho, Young Deuk Choi, Suki Kang, Bora Kim, Kook Jin Kim, Eun Ji Lim, Sun Young Rha, and et al. 2020. "Scale-Up Evaluation of a Composite Tumor Marker Assay for the Early Detection of Renal Cell Carcinoma" Diagnostics 10, no. 10: 750. https://doi.org/10.3390/diagnostics10100750