A Laboratory-Friendly CTC Identification: Comparable Double-Immunocytochemistry with Triple-Immunofluorescence

Area of Science:

• Oncology
• Biotechnology
• Medical Diagnostics

Background:

• Circulating tumor cells (CTCs) in blood are crucial for cancer metastasis and treatment resistance, impacting 90% of cancer deaths.
• Identifying CTCs offers significant predictive and prognostic value in solid tumors.
• Current CTC enumeration is not standard practice due to a lack of convenient, cost-effective methods.

Purpose of the Study:

• To present a specific, sensitive, and laboratory-friendly method for simultaneous isolation and identification of CTCs.
• To address the need for a cost-effective and convenient CTC detection technique for routine clinical use.

Main Methods:

• Developed a parallel double-detection format using size-based isolation via a pressure-guided method on microfilters.
• Employed dual identification strategies: double-immunocytochemistry alongside standard triple-immunofluorescence.
• Utilized epithelial markers (EpCAM, CK8,18,19) and a white blood cell exclusion marker (CD45).

Main Results:

• Successfully isolated and identified CTCs from 91 patients with diverse solid tumors (lung, endometrium, ovary, esophagus, prostate, liver).
• Validated the method's specificity using spiked samples, healthy donor blood, leucopaks, and blood from metastatic patients.
• Demonstrated a user-friendly and cost-effective approach for CTC detection.

Conclusions:

• The presented parallel double-detection method offers a specific, sensitive, and practical solution for CTC isolation and identification.
• This cost-effective technique has the potential to facilitate routine CTC monitoring in community cancer centers.
• Regular CTC detection can aid in prognosis and treatment guidance before and after surgery.