Limits...
Comparison of three molecular assays for the detection and molecular characterization of circulating tumor cells in breast cancer.

Strati A, Kasimir-Bauer S, Markou A, Parisi C, Lianidou ES - Breast Cancer Res. (2013)

Bottom Line: The concordance between the AdnaTest and CK-19 RT-qPCR was 70.6% while between the AdnaTest and multiplex RT-qPCR was 68.6%.Better agreement was found in the metastatic setting, possibly explained by the higher tumor load in this group.Our results indicate the importance of CTC heterogeneity for their detection by different analytical methodologies.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Introduction: Comparison studies between different analytical methodologies for circulating tumor cells (CTC) detection and molecular characterization are urgently needed, since standardization of assays is essential before their use in clinical practice.

Methods: We compared three different CTC molecular assays. To avoid discrepancies due to pre-analytical errors we used the same cDNAs throughout our study. CTC were isolated using anti-EpCAM and anti-MUC1 coated magnetic beads from 2 × 5 ml of peripheral blood of 254 early and 51 metastatic breast cancer patients and 30 healthy individuals. The same cDNAs were analyzed by: a) singleplex RT-qPCR assay for CK-19; b) multiplex RT-qPCR for CK-19, HER-2, MAGE- A3, and PBGD; and c) a commercially available molecular assay (AdnaTest BreastCancer) for GA733-2, MUC-1, HER-2 and beta-actin.

Results: In early breast cancer, CK-19 RT-qPCR, multiplex RT-qPCR and the AdnaTest, were positive for the presence of CTC in 14.2%, 22.8% and 16.5% subjects, respectively. The concordance between the AdnaTest and CK-19 RT-qPCR was 72.4% while between the AdnaTest and multiplex RT-qPCR was 64.6%. In patients with overt metastasis, CK-19 RT-qPCR, multiplex RT-qPCR and the AdnaTest were positive in 41.2%, 39.2% and 54.9% patients, respectively. The concordance between the AdnaTest and CK-19 RT-qPCR was 70.6% while between the AdnaTest and multiplex RT-qPCR was 68.6%.

Conclusions: All CTC assays gave similar results in about 70% of cases. Better agreement was found in the metastatic setting, possibly explained by the higher tumor load in this group. Discordances could be attributed to the different gene transcripts used to evaluate CTC positivity. Our results indicate the importance of CTC heterogeneity for their detection by different analytical methodologies.

Show MeSH

Related in: MedlinePlus

Molecular profiling of CTC in early breast cancer (n = 254). Red and green indicates positive and negative detection, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3672668&req=5

Figure 1: Molecular profiling of CTC in early breast cancer (n = 254). Red and green indicates positive and negative detection, respectively.

Mentions: In early breast cancer, we found an agreement between the AdnaTest BreastCancer™ and CK-19 RT-qPCR in 184/254 (72.4%) of the cases. However, the correlation concerning the CTC positivity between these two assays was not significant (P = 0.344), since there were 32 samples that were found positive for CTC by the CK-19 assay but negative by the AdnaTest and 38 samples found positive for CTC by the AdnaTest but negative by the CK-19 assay. The concordance between the AdnaTest and multiplex RT-qPCR in this group of patients was 64.6%. There was no significant correlation (P = 0.065) in this case as well, since there were 53 samples found positive for CTC by the multiplex RT-qPCR assay but negative by the AdnaTest and 37 samples found positive for CTC by the AdnaTest but negative by the multiplex RT-qPCR assay (Table 2). In this group, all combinations found for each molecular target are shown in Figure 1.


Comparison of three molecular assays for the detection and molecular characterization of circulating tumor cells in breast cancer.

Strati A, Kasimir-Bauer S, Markou A, Parisi C, Lianidou ES - Breast Cancer Res. (2013)

Molecular profiling of CTC in early breast cancer (n = 254). Red and green indicates positive and negative detection, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3672668&req=5

Figure 1: Molecular profiling of CTC in early breast cancer (n = 254). Red and green indicates positive and negative detection, respectively.
Mentions: In early breast cancer, we found an agreement between the AdnaTest BreastCancer™ and CK-19 RT-qPCR in 184/254 (72.4%) of the cases. However, the correlation concerning the CTC positivity between these two assays was not significant (P = 0.344), since there were 32 samples that were found positive for CTC by the CK-19 assay but negative by the AdnaTest and 38 samples found positive for CTC by the AdnaTest but negative by the CK-19 assay. The concordance between the AdnaTest and multiplex RT-qPCR in this group of patients was 64.6%. There was no significant correlation (P = 0.065) in this case as well, since there were 53 samples found positive for CTC by the multiplex RT-qPCR assay but negative by the AdnaTest and 37 samples found positive for CTC by the AdnaTest but negative by the multiplex RT-qPCR assay (Table 2). In this group, all combinations found for each molecular target are shown in Figure 1.

Bottom Line: The concordance between the AdnaTest and CK-19 RT-qPCR was 70.6% while between the AdnaTest and multiplex RT-qPCR was 68.6%.Better agreement was found in the metastatic setting, possibly explained by the higher tumor load in this group.Our results indicate the importance of CTC heterogeneity for their detection by different analytical methodologies.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Introduction: Comparison studies between different analytical methodologies for circulating tumor cells (CTC) detection and molecular characterization are urgently needed, since standardization of assays is essential before their use in clinical practice.

Methods: We compared three different CTC molecular assays. To avoid discrepancies due to pre-analytical errors we used the same cDNAs throughout our study. CTC were isolated using anti-EpCAM and anti-MUC1 coated magnetic beads from 2 × 5 ml of peripheral blood of 254 early and 51 metastatic breast cancer patients and 30 healthy individuals. The same cDNAs were analyzed by: a) singleplex RT-qPCR assay for CK-19; b) multiplex RT-qPCR for CK-19, HER-2, MAGE- A3, and PBGD; and c) a commercially available molecular assay (AdnaTest BreastCancer) for GA733-2, MUC-1, HER-2 and beta-actin.

Results: In early breast cancer, CK-19 RT-qPCR, multiplex RT-qPCR and the AdnaTest, were positive for the presence of CTC in 14.2%, 22.8% and 16.5% subjects, respectively. The concordance between the AdnaTest and CK-19 RT-qPCR was 72.4% while between the AdnaTest and multiplex RT-qPCR was 64.6%. In patients with overt metastasis, CK-19 RT-qPCR, multiplex RT-qPCR and the AdnaTest were positive in 41.2%, 39.2% and 54.9% patients, respectively. The concordance between the AdnaTest and CK-19 RT-qPCR was 70.6% while between the AdnaTest and multiplex RT-qPCR was 68.6%.

Conclusions: All CTC assays gave similar results in about 70% of cases. Better agreement was found in the metastatic setting, possibly explained by the higher tumor load in this group. Discordances could be attributed to the different gene transcripts used to evaluate CTC positivity. Our results indicate the importance of CTC heterogeneity for their detection by different analytical methodologies.

Show MeSH
Related in: MedlinePlus