ReplicationCompetentVirus_RCV_QualityChecker

Replication Competent Virus RCV

Huvac human vaccines vaccination adjuvants mRNA vaccines sterility endotoxins cold chain

Utility description: Replication Competent Virus RCV

Replication Competent Virus RCV Quality Checker — RCV Control in Gene Therapy

ℹ️ Utility performs critical safety control of viral vectors, detecting presence of Replication Competent Viruses (RCV) according to FDA and EMA guidelines:
• Sensitive Analysis: Processing of PCR (qPCR/ddPCR) or biological test data.
• Limit of Detection (LoD): Comparison of result with validated method sensitivity threshold.
• System Suitability: Mandatory check of positive and negative control correctness.
• Patient Safety: Exclusion of uncontrolled viral infection risk.

⚠️ IMPORTANT:
• RCV form rarely, but their consequences can be fatal.
• Test must have high sensitivity level (low LoD).

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

Input format:
BatchNumber,VectorType,RCV_Copies_Per_mL,LoD_Copies_Per_mL,Max_Allowed_RCV,PositiveControl_Valid,NegativeControl_Valid,DetectionMethod

Example:
LV-001,Lentivirus,0.0,10.0,1.0,true,true,qPCR

📍 Scope of Application (Usage Where):
• Gene Therapy Production: Release of lentiviral, retroviral and AAV vector batches.
• Vaccine Development: Control of viral vector vaccines.
• Regulatory Dossier: Preparation of safety data for IND/BLA.
• Laboratory Control: Monitoring of master and working cell banks.

— WHY IS THIS NEEDED?
Recombinant viral vectors are created by splitting virus genome into several plasmids.
During production, reverse recombination is possible, leading to appearance of full-fledged virus capable of replication.
Strict RCV control guarantees that product will deliver gene only once and will not cause infectious process.

⚠️ CRITICAL:
• LoD: Method must be validated for detection of single RCV copies.
• Controls: Failure of positive control means false-negative result (unacceptable).
• Zero Tolerance: For most clinical applications, only "Not Detected" status is allowed.
• Sample Volume: Sufficient sample volume is necessary for statistical reliability.

Key features:
• Automatic test validity check
• Comparison with LoD and absolute limits
• Support for various detection methods
• Report generation for regulatory authorities
• Compliance with GMP requirements for Advanced Therapy Medicinal Products (ATMP)

Critical parameters:
• RCV Copies: Must be < LoD
• Positive Control: Valid
• Negative Control: Valid
• LoD: Validated value

💡 Usage tips:
1. Method Validation: Regularly confirm LoD using RCV standards.
2. Contamination Prevention: Strict separation of pre- and post-PCR zones.
3. Sample Volume: Use maximum possible volume of DNA/RNA extract for analysis.
4. Documentation: Save raw amplification data (Ct values) together with report.
5. Investigation: Any positive signal requires immediate investigation and retest.

⚠️ Note: This utility is a safety control tool. Positive RCV result automatically blocks batch release and requires deep investigation of contamination source.

input.csv

BatchNumber,VectorType,RCV_Copies_Per_mL,LoD_Copies_Per_mL,Max_Allowed_RCV,PositiveControl_Valid,NegativeControl_Valid,DetectionMethod
LV-2026-CAR-T-001,Lentivirus,0.0,10.0,1.0,true,true,qPCR
AAV-2026-GT-002,AAV9,55.0,10.0,1.0,true,true,ddPCR
ADV-2026-VAC-003,Adenovirus,0.0,5.0,1.0,true,true,Cell-Assay

URS & FS — User Requirements and Functional Specification

This document describes the controlled interface and behaviour of ReplicationCompetentVirus_RCV_QualityChecker for Replication Competent Virus RCV Quality 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.

ℹ️ Utility performs critical safety control of viral vectors, detecting presence of Replication Competent Viruses (RCV) according to FDA and EMA guidelines:
• Limit of Detection (LoD): Comparison of result with validated method sensitivity threshold.
• Test must have high sensitivity level (low LoD).
⚠️ CRITICAL:
• LoD: Method must be validated for detection of single RCV copies.
• Comparison with LoD and absolute limits
• Compliance with GMP requirements for Advanced Therapy Medicinal Products (ATMP)
Critical parameters:
• RCV Copies: Must be < LoD

URS — User Requirements Specification

ID	Requirement	Criticality	Acceptance criterion
URS-001	The utility shall accept an input.csv file for Replication Competent Virus RCV Quality 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	`BatchNumber`	string / controlled vocabulary	`LV-2026-CAR-T-001`	Batch or lot identifier used for traceability.
2	`VectorType`	string / controlled vocabulary	`Lentivirus`	Controlled input parameter for deterministic QC rules.
3	`RCV_Copies_Per_mL`	decimal	`0.0`	Volume/dose used for load, limit or release calculation.
4	`LoD_Copies_Per_mL`	decimal	`10.0`	Volume/dose used for load, limit or release calculation.
5	`Max_Allowed_RCV`	decimal	`1.0`	Controlled input parameter for deterministic QC rules.
6	`PositiveControl_Valid`	string / controlled vocabulary	`true`	Control point/control sample for assay run validity.
7	`NegativeControl_Valid`	string / controlled vocabulary	`true`	Control point/control sample for assay run validity.
8	`DetectionMethod`	string / controlled vocabulary	`qPCR`	Method or process approach; controlled reference value.

BatchNumber,VectorType,RCV_Copies_Per_mL,LoD_Copies_Per_mL,Max_Allowed_RCV,PositiveControl_Valid,NegativeControl_Valid,DetectionMethod
LV-2026-CAR-T-001,Lentivirus,0.0,10.0,1.0,true,true,qPCR
AAV-2026-GT-002,AAV9,55.0,10.0,1.0,true,true,ddPCR
ADV-2026-VAC-003,Adenovirus,0.0,5.0,1.0,true,true,Cell-Assay

Input validation rules

ID	Field	Rule	Criticality
VR-001	`BatchNumber`	The field shall match an approved dictionary or accepted string representation.	High
VR-002	`VectorType`	The field shall match an approved dictionary or accepted string representation.	High
VR-003	`RCV_Copies_Per_mL`	The field shall match an approved dictionary or accepted string representation.	High
VR-004	`LoD_Copies_Per_mL`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-005	`Max_Allowed_RCV`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-006	`PositiveControl_Valid`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-007	`NegativeControl_Valid`	The field shall match an approved dictionary or accepted string representation.	Medium
VR-008	`DetectionMethod`	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 Replication Competent Virus RCV Quality 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": "replicationcompetentvirus-rcv-qualitychecker",
  "utilityFolder": "ReplicationCompetentVirus_RCV_QualityChecker",
  "package": "huvac",
  "overallStatus": "PASS|WARNING|FAIL",
  "sourceFile": "input.csv",
  "processedAtUtc": "2026-06-10T00:00:00Z",
  "checks": [
    {
      "parameter": "BatchNumber",
      "value": "LV-2026-CAR-T-001",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-001"
    },
    {
      "parameter": "VectorType",
      "value": "Lentivirus",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-002"
    },
    {
      "parameter": "RCV_Copies_Per_mL",
      "value": "0.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-003"
    },
    {
      "parameter": "LoD_Copies_Per_mL",
      "value": "10.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-004"
    },
    {
      "parameter": "Max_Allowed_RCV",
      "value": "1.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-005"
    },
    {
      "parameter": "PositiveControl_Valid",
      "value": "true",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-006"
    },
    {
      "parameter": "NegativeControl_Valid",
      "value": "true",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-007"
    },
    {
      "parameter": "DetectionMethod",
      "value": "qPCR",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-008"
    }
  ],
  "criticalFindings": [],
  "warnings": [],
  "audit": {
    "inputHash": "sha256:<calculated at runtime>",
    "rulesVersion": "<utility executable version>",
    "documentation": "ReplicationCompetentVirus_RCV_QualityChecker.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

Biopharmaceuticals Extended QC Suite

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QC utility package for infectious diseases: antibacterial, antiviral, antifungal and antiparasitic products, microbiology, sterility, endotoxins, viral safety and vaccine release checks.

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