Quality control in nucleic acid testing--where do we stand?

Area of Science:

• Clinical Diagnostics
• Molecular Biology
• Virology

Background:

• Nucleic acid amplification techniques (NATs) are crucial for clinical diagnosis.
• Early NAT implementation faced significant issues with specificity, sensitivity, and quantitative variability.
• Key challenges included reagent standardization, contamination control, and genotype-independent detection.

Purpose of the Study:

• To assess the progress and challenges in quality control for NATs in clinical diagnostics.
• To evaluate the impact of external quality control initiatives on NAT performance.
• To identify remaining areas for improvement in NAT accuracy and reliability.

Main Methods:

• Review of progress in addressing identified NAT problem areas.
• Establishment and operation of the European Union Quality Control Concerted Action for Nucleic Acid Amplification in Diagnostic Virology.
• Distribution of 14 proficiency panels with simulated clinical samples to numerous international laboratories for testing.

Main Results:

• Dramatic improvement in specificity, with false-positive rates below 3% for most panels.
• Significant contributions from increased laboratory expertise, contamination control, and standardized reagents.
• Persistent challenges with sensitivity, showing false-negative rates up to 50% in some cases.

Conclusions:

• Quality control for NATs in clinical diagnosis has seen considerable progress, particularly in specificity.
• Ongoing challenges in sensitivity and standardization of quantitation require further attention.
• Continuous quality control efforts are essential for reliable NAT-based diagnostics.