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Comparison of assay methods for detection of circulating tumor cells in metastatic breast cancer: AdnaGen AdnaTest BreastCancer Select/Detect™ versus Veridex CellSearch™ system.

Author(s): Andreopoulou E, Yang LY, Rangel KM, Reuben JM, Hsu L, Krishnamurthy S, Valero V, Fritsche HA, Cristofanilli M

Publication: Int J Cancer, 2012, Vol. 130, Page 1590-7

PubMed ID: 21469140 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study compared the sensitivity of two difference assays for enumeration of circulating tumor cells (CTCs): the CellSearch System and AdnaTest Breast Cancer Select/Detect System. Peripheral blood (10 mL) from 55 metastatic breast cancer patients was collected into CellSave tubes, maintained at room temperature, and processed within 72 h using the CellSearch System or collected in EDTA tubes (5 mL) for the AdnaTest in which tumor cells were enriched using immunomagnetic beads; mRNA was isolated and transcribed into cDNA; and then MUC-1, HER2, and GA733-2 expression was detected in a two-step real-time qRT-PCR assay. For the CellSearch System, two cutoff values for a CTC-positive specimen were evaluated: ≥2 and ≥5 CTCs per 7.5 ml of blood. For the AdnaTest, a specimen was regarded as CTC positive if at least one of the three PCR markers was detected at a concentration of ≥0.15 ng/µl. Immunohistochemistry for estrogen receptor (ER) and progesterone receptor (PR) was performed on 5 µm-thick tissue sections of the primary tumor and for HER2 status using 4 µm-thick sections. HER status was also confirmed by fluorescence in situ hybridization (FISH) analysis.

   Summary of Findings:

   The percentage of patients that were classified as CTC-positive using the CellSearch system was 47% (26/55) when a cutoff value of ≥2 CTCs/7.5 ml of blood was applied and 36% (20/55) with a cutoff value of ≥5 CTCs/ 7.5 ml of blood. Comparable results were obtained using the AdnaTest method as 53% of patients (29/55) were classified as CTC-positive (18% positive for GA733-2, 44% for MUC-1, and 35% for HER2) with 19 of these being classified as positive for either two or all three genes and the remaining 10 specimens were positive for only one marker. Concordance between the AdnaTest and the CellSearch System was 73% (cutoff value ≥2 CTCs/7.5 ml of blood) and 69% (cutoff value ≥5 CTCs/7.5 ml of blood). Of the 55 primary tumors evaluated, hormone receptor status was positive for ER/PR overexpression in 36 patients (65%) as detected by IHC, HER2 positive in 12 patients (22%) as detected by IHC and FISH, and triple-negative (negative for ER, PR, and HER2) in 11 patients (20%). Although 19 patients were classified as HER2 positive when blood specimens were analyzed by the AdnaTest, IHC/FISH analysis of the primary tumor revealed concordant findings in only 5 patients, discordant findings in 11 patients, and an undetermined status in the remaining three patients. CTC detection method performance was not associated with ER, PR, or HER2 status of the primary tumor.

   Biospecimens

   • Bodily Fluid - Blood
   • Tissue - Breast

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Fluorescent microscopy
   RNA	In situ hybridization
   Protein	Immunohistochemistry
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Real-time qRT-PCR Specific	Targeted nucleic acid	MUC-1 HER2 GA733-2
   Biospecimen Aliquots and Components	Cell capture method	CellSave AdnaTest
   In situ hybridization Specific	Targeted nucleic acid	HER2
   Immunohistochemistry Specific	Targeted peptide/protein	estrogen receptor (ER) progesterone receptor (PR) HER2
   Biospecimen Aliquots and Components	Cell number	≥2 CTCs/7.5 ml of blood ≥5 CTCs/7.5 ml of blood

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