Laboratory Optimization Tweaks for Sanger Sequencing in a Resource-Limited Setting

Area of Science:

• Molecular Diagnostics
• Virology
• Clinical Laboratory Science

Background:

• High-tech molecular methods face implementation challenges in resource-limited settings.
• Genotypic HIV drug resistance testing is crucial for effective treatment.
• Accreditation requires robust and validated laboratory processes.

Purpose of the Study:

• To describe the optimization of a laboratory workflow for genotypic HIV drug resistance testing.
• To achieve International Organization for Standardization (ISO) 15189:2012 accreditation in a resource-limited setting.
• To demonstrate the efficiency and quality of the optimized workflow.

Main Methods:

• Optimization of laboratory processes including sample preparation, RT-PCR, and cycle sequencing.
• Implementation of stringent quality control measures.
• Participation in external quality assurance programs.

Main Results:

• Achieved ISO 15189:2012 accreditation for genotypic HIV drug resistance testing.
• Recorded high sequencing success rates and good quality sequence data.
• Scored 100% in an external quality assurance program evaluating mutation points.

Conclusions:

• Optimized laboratory workflows can overcome challenges in resource-limited settings.
• The described workflow maximizes output, minimizes reagent wastage, and ensures data quality.
• This approach supports laboratories in achieving accurate results and reducing costs.