CtcEnumerationQcChecker

Ctc Enumeration QC

Liquid Biopsy жидкостная биопсия cfDNA ctDNA CTC exosomes NGS qPCR

Utility description: Ctc Enumeration QC

Ctc Enumeration QC Checker — Circulating Tumor Cell Enumeration Quality Control

ℹ️ Utility performs comprehensive QC verification of CTC analysis according to ISHDE standards and FDA guidance:
• Spike-In Recovery: Capture efficiency control using labeled cell lines.
• WBC Contamination: Assessment of leukocyte impurity creating false-positive signals.
• Blood Volume Sufficiency: Verification of adequate blood volume for required sensitivity.
• Phenotype Validation: Verification of classic CTC phenotype (DAPI+/CK+/CD45-/morphology).
• Background Noise: Assessment of non-specific signal complicating identification.

⚠️ IMPORTANT:
• CTC exist at 1–10 cells per 7.5 mL blood — loss of even one cell is significant.
• CD45+ cells are NOT CTC and must be excluded from count.
• Low recovery rate makes negative result unreliable (possible false-negative).

Usage:
CtcEnumerationQcChecker.exe → demo mode (console output)
CtcEnumerationQcChecker.exe input.csv output.json → evaluate your data

Input format:
SampleID,Platform,BloodVolumeProcessed_mL,MinBloodVolume_mL,SpikeInCells_Added,SpikeInCells_Recovered,MinRecoveryRate_Percent,WBC_Contamination_Count,MaxWBC_Contamination,CTC_Count_Raw,ClinicalThreshold,MorphologyIntact,DAPI_Positive,EpCAM_CK_Positive,CD45_Negative,BackgroundNoise_Score,MaxBackgroundNoise

Example:
CTC-001,CellSearch,7.5,7.0,100,82,70,15,100,7,5,true,true,true,true,1.5,3.0

📍 Scope of Application (Usage Where):
• Clinical Laboratories: QC before CTC count result release.
• Oncology Research: Therapy response monitoring via CTC dynamics.
• CTC Platform Development: Validation of new enrichment and detection methods.
• Regulatory Submissions: Documentation of CTC test analytical validity.

— WHY IS THIS NEEDED?
CTC enumeration is used for therapy change decisions in metastatic cancer.
Technical artifacts (cell loss, WBC contamination, poor morphology) directly impact clinical decisions.
Automated QC eliminates subjectivity of manual assessment and guarantees every counted CTC is a true tumor cell.

⚠️ CRITICAL:
• Spike-In Recovery: <70% = unacceptable for quantitative analysis.
• CD45 Negativity: CD45+ = leukocyte, NOT CTC. Exclude from count.
• Blood Volume: <7.0 mL reduces analytical sensitivity below clinical threshold.
• Morphology: Damaged cells cannot be reliably identified as CTC.
• Background: High noise makes counting subjective and irreproducible.

Key features:
• Five-parameter CTC analysis quality assessment
• Automatic CTC phenotype verification
• Integration of spike-in recovery and WBC contamination
• Status classification (Pass/Review/Fail)
• Compliance with ISHDE consensus guidelines

Critical parameters:
• Spike-In Recovery: ≥ 70%
• WBC Contamination: ≤ Max Limit
• Blood Volume: ≥ 7.0 mL
• Phenotype: DAPI+/CK+/CD45-/Intact
• Background Noise: ≤ Max Score

💡 Usage tips:
1. Spike-In Control: Add labeled cells BEFORE enrichment step to monitor entire process.
2. Dual Staining: Always use CD45 to exclude leukocytes.
3. Morphology: Train operators on intact cellular morphology recognition.
4. Blood Volume: Strictly maintain minimum 7.5 mL for standardized thresholds.
5. Trends: Monitor recovery rate over time to detect reagent/cartridge degradation.

⚠️ Note: This utility is an analytical QC tool. It does not replace clinical interpretation by oncologist but guarantees that CTC count in report is technically reliable.

input.csv

SampleID,Platform,BloodVolumeProcessed_mL,MinBloodVolume_mL,SpikeInCells_Added,SpikeInCells_Recovered,MinRecoveryRate_Percent,WBC_Contamination_Count,MaxWBC_Contamination,CTC_Count_Raw,ClinicalThreshold,MorphologyIntact,DAPI_Positive,EpCAM_CK_Positive,CD45_Negative,BackgroundNoise_Score,MaxBackgroundNoise
CTC-2026-001,CellSearch,7.5,7.0,100,82,70.0,15,100,7,5.0,true,true,true,true,1.5,3.0
CTC-2026-002,Parsortix,5.0,7.0,100,45,70.0,350,100,3,5.0,false,true,true,false,6.5,3.0
CTC-2026-003,CTC-iChip,7.5,7.0,50,42,70.0,8,50,0,3.0,true,true,true,true,0.8,3.0

URS & FS — User Requirements and Functional Specification

This document describes the controlled interface and behaviour of CtcEnumerationQcChecker for Ctc Enumeration QC Checker.

Domain limits URS input.csv FS IQ/OQ/PQ

Domain limits and critical parameters

Key fragments from the source description are shown below. Before production use, limits must be verified against the approved specification, registration dossier and local SOPs.

• WBC Contamination: Assessment of leukocyte impurity creating false-positive signals.
• CD45+ cells are NOT CTC and must be excluded from count.
• CTC Platform Development: Validation of new enrichment and detection methods.
⚠️ CRITICAL:
• Spike-In Recovery: <70% = unacceptable for quantitative analysis.
• Blood Volume: <7.0 mL reduces analytical sensitivity below clinical threshold.
Critical parameters:
• Spike-In Recovery: ≥ 70%
• WBC Contamination: ≤ Max Limit
• Blood Volume: ≥ 7.0 mL
• Background Noise: ≤ Max Score
1. Spike-In Control: Add labeled cells BEFORE enrichment step to monitor entire process.

URS — User Requirements Specification

ID	Requirement	Criticality	Acceptance criterion
URS-001	The utility shall accept an input.csv file for Ctc Enumeration QC Checker with headers defined in the data contract.	High	The file is processed without manual header editing.
URS-002	The utility shall perform deterministic QC evaluation without machine learning and without probabilistic conformance decisions.	High	Identical input data, rule version and configuration produce reproducible results.
URS-003	The utility shall validate mandatory fields, data types, ranges, units and domain plausibility.	High	Schema, conversion and range errors are explicitly reported.
URS-004	The utility shall apply domain limits and rules from the description, approved specification, registration dossier and local SOPs.	High	Each check has PASS/WARNING/FAIL and a clear message.
URS-005	The utility shall generate output.json with machine-readable results, source values, warnings, failures and critical findings.	High	JSON is suitable for LIMS/ELN/MES integration and QA/QC review.
URS-006	The utility shall preserve traceability between batch/sample, input file, applied rules and final status.	High	Output contains identifiers, checked parameters and audit metadata.
URS-007	The documentation shall support IQ/OQ/PQ, CSV/CSA and review by internal QA or inspectors.	Medium	URS, FS, input/output contract and test scenarios are supplied with the utility.
URS-008	The utility shall be used as a QC decision-support tool and not as a substitute for approved specifications and QA/QP release decision.	Medium	Documentation states change control and limit-verification expectations.

input.csv contract

#	Field	Type	Sample	Purpose
1	`SampleID`	string / controlled vocabulary	`CTC-2026-001`	Sample or laboratory specimen identifier.
2	`Platform`	string / controlled vocabulary	`CellSearch`	Controlled input parameter for deterministic QC rules.
3	`BloodVolumeProcessed_mL`	decimal	`7.5`	Volume/dose used for load, limit or release calculation.
4	`MinBloodVolume_mL`	decimal	`7.0`	Volume/dose used for load, limit or release calculation.
5	`SpikeInCells_Added`	decimal	`100`	Controlled input parameter for deterministic QC rules.
6	`SpikeInCells_Recovered`	decimal	`82`	Controlled input parameter for deterministic QC rules.
7	`MinRecoveryRate_Percent`	decimal	`70.0`	Recovery; indicator of analytical validity.
8	`WBC_Contamination_Count`	integer / decimal	`15`	Count parameter used for microbiological, particulate or cellular control.
9	`MaxWBC_Contamination`	integer / decimal	`100`	Controlled input parameter for deterministic QC rules.
10	`CTC_Count_Raw`	integer / decimal	`7`	Count parameter used for microbiological, particulate or cellular control.
11	`ClinicalThreshold`	decimal	`5.0`	Controlled input parameter for deterministic QC rules.
12	`MorphologyIntact`	string / controlled vocabulary	`true`	pH; parameter relevant to stability, compatibility and batch suitability.
13	`DAPI_Positive`	string / controlled vocabulary	`true`	Controlled input parameter for deterministic QC rules.
14	`EpCAM_CK_Positive`	string / controlled vocabulary	`true`	Controlled input parameter for deterministic QC rules.
15	`CD45_Negative`	string / controlled vocabulary	`true`	Controlled input parameter for deterministic QC rules.
16	`BackgroundNoise_Score`	decimal	`1.5`	Controlled input parameter for deterministic QC rules.
17	`MaxBackgroundNoise`	decimal	`3.0`	Controlled input parameter for deterministic QC rules.

SampleID,Platform,BloodVolumeProcessed_mL,MinBloodVolume_mL,SpikeInCells_Added,SpikeInCells_Recovered,MinRecoveryRate_Percent,WBC_Contamination_Count,MaxWBC_Contamination,CTC_Count_Raw,ClinicalThreshold,MorphologyIntact,DAPI_Positive,EpCAM_CK_Positive,CD45_Negative,BackgroundNoise_Score,MaxBackgroundNoise
CTC-2026-001,CellSearch,7.5,7.0,100,82,70.0,15,100,7,5.0,true,true,true,true,1.5,3.0
CTC-2026-002,Parsortix,5.0,7.0,100,45,70.0,350,100,3,5.0,false,true,true,false,6.5,3.0
CTC-2026-003,CTC-iChip,7.5,7.0,50,42,70.0,8,50,0,3.0,true,true,true,true,0.8,3.0

Input validation rules

ID	Field	Rule	Criticality
VR-001	`SampleID`	The field shall match an approved dictionary or accepted string representation.	High
VR-002	`Platform`	The field shall match an approved dictionary or accepted string representation.	High
VR-003	`BloodVolumeProcessed_mL`	The field shall match an approved dictionary or accepted string representation.	High
VR-004	`MinBloodVolume_mL`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-005	`SpikeInCells_Added`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-006	`SpikeInCells_Recovered`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-007	`MinRecoveryRate_Percent`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-008	`WBC_Contamination_Count`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-009	`MaxWBC_Contamination`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-010	`CTC_Count_Raw`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-011	`ClinicalThreshold`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-012	`MorphologyIntact`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-013	`DAPI_Positive`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-014	`EpCAM_CK_Positive`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-015	`CD45_Negative`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-016	`BackgroundNoise_Score`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-017	`MaxBackgroundNoise`	The field shall match an approved dictionary or accepted string representation.	Medium

FS — Functional Specification

ID	Function	Implementation
FS-001	CLI execution	Support execution modes: demo mode without arguments and production mode input.csv output.json.
FS-002	CSV import	Read input.csv in UTF-8/CSV-compatible format and validate header and expected columns.
FS-003	Schema validation	Check mandatory fields, column count, unknown key fields and empty mandatory values.
FS-004	Type conversion	Convert numeric, flag and text values; invalid format is recorded as a row-level error.
FS-005	Domain rule engine	Apply rules for Ctc Enumeration QC Checker, including critical limits from the description and approved specification.
FS-006	Status aggregation	Produce final status: FAIL for critical failure, WARNING for non-critical deviation, PASS for conformance.
FS-007	JSON export	Write output.json with detailed checks, source values, warnings, failures and critical findings.
FS-008	Audit support	Keep result structure suitable for review, deviation investigation and calculation reproduction.
FS-009	Integration contract	Support the scenario LIMS/ELN/MES → input.csv → utility → output.json → portal/admin review.
FS-010	Error handling	Return explicit messages for missing file, empty CSV, invalid schema, output write failure and invalid format.

Example output.json

{
  "utilityId": "ctcenumerationqcchecker",
  "utilityFolder": "CtcEnumerationQcChecker",
  "package": "LiquidBiopsy",
  "overallStatus": "PASS|WARNING|FAIL",
  "sourceFile": "input.csv",
  "processedAtUtc": "2026-06-10T00:00:00Z",
  "checks": [
    {
      "parameter": "SampleID",
      "value": "CTC-2026-001",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-001"
    },
    {
      "parameter": "Platform",
      "value": "CellSearch",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-002"
    },
    {
      "parameter": "BloodVolumeProcessed_mL",
      "value": "7.5",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-003"
    },
    {
      "parameter": "MinBloodVolume_mL",
      "value": "7.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-004"
    },
    {
      "parameter": "SpikeInCells_Added",
      "value": "100",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-005"
    },
    {
      "parameter": "SpikeInCells_Recovered",
      "value": "82",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-006"
    },
    {
      "parameter": "MinRecoveryRate_Percent",
      "value": "70.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-007"
    },
    {
      "parameter": "WBC_Contamination_Count",
      "value": "15",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-008"
    },
    {
      "parameter": "MaxWBC_Contamination",
      "value": "100",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-009"
    },
    {
      "parameter": "CTC_Count_Raw",
      "value": "7",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-010"
    },
    {
      "parameter": "ClinicalThreshold",
      "value": "5.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-011"
    },
    {
      "parameter": "MorphologyIntact",
      "value": "true",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-012"
    }
  ],
  "criticalFindings": [],
  "warnings": [],
  "audit": {
    "inputHash": "sha256:<calculated at runtime>",
    "rulesVersion": "<utility executable version>",
    "documentation": "CtcEnumerationQcChecker.documentation.html"
  }
}

Traceability matrix

URS	FS	Test	Evidence
URS-001	FS-001, FS-002	OQ-001	Verify execution and import of valid input.csv.
URS-002	FS-005, FS-006	OQ-004	Repeat the same dataset and compare output.json.
URS-003	FS-003, FS-004, FS-010	OQ-002, OQ-003	Verify missing columns and invalid types.
URS-004	FS-005, FS-006	OQ-004, PQ-001	Verify critical deviations on real/boundary data.
URS-005	FS-007, FS-009	OQ-005	Verify JSON schema and downstream-system suitability.
URS-006	FS-008	OQ-006	Verify identifiers and audit metadata.
URS-007	FS-008, FS-010	IQ-001, OQ-007	Verify documentation completeness and control evidence.
URS-008	FS-005, FS-008	PQ-002	Verify review workflow and no replacement of QA decision.

IQ/OQ/PQ test scenarios

ID	Scenario	Expected result
IQ-001	Verify executable, input.csv, documentation and checksum availability.	Delivery set is complete; version is recorded.
OQ-001	Valid sample row from input.csv.	PASS or acceptable WARNING according to rules.
OQ-002	Remove a mandatory CSV column.	Schema error or FAIL with missing-column reference.
OQ-003	Place a non-numeric value into a numeric field.	Type-conversion error with row/field reference.
OQ-004	Set a critical parameter outside the limit.	FAIL and critical finding.
OQ-005	Verify output.json structure.	All mandatory sections are present and JSON is valid.
OQ-006	Verify batch/sample traceability.	Input and result identifiers match.
PQ-001	Verify 3–5 real user batches/samples.	Result is confirmed by QC/QA review.
PQ-002	Verify deviation workflow and manual QA decision.	Utility supports review but does not replace approved decision.

QA/QC and change control

Do not rename columns without updating validator, documentation and test set.
Retain input.csv, output.json, executable version and checksum.
Before production use, perform IQ/OQ/PQ or equivalent CSV/CSA verification.
Critical limits shall be verified against the approved specification, registration dossier and local SOPs.
The utility provides structured QC decision support; final release decision remains with QA/QP and approved procedures.

Included in packages

Liquid Biopsy QC Suite

QC and pre-analytical control package for liquid biopsy workflows: cfDNA/ctDNA, CTC, EV/exosomes, methylation, NGS/qPCR/ddPCR, sample quality, contamination, sensitivity and reporting checks.

Open