Cell-based reference samples designed with specific differences in microRNA biomarkers

Area of Science:

• Biotechnology
• Molecular Biology
• Genomics

Background:

• Clinical laboratories require proficiency testing for quality assurance, unlike research laboratories.
• Developing reliable reference samples is crucial for assessing measurement processes and detecting true biological differences.
• Cell-based reference samples offer a method to create artificial biomarker signatures for laboratory evaluation.

Purpose of the Study:

• To demonstrate the feasibility of creating cell-based reference samples for laboratory measurement assessment.
• To evaluate the utility of these reference samples in detecting true differences between biological samples.
• To establish a paradigm for creating and using reference samples in research and clinical laboratory settings.

Main Methods:

• Utilized lung cancer-associated cell lines with distinct microRNA (miRNA) profiles.
• Employed reverse transcription polymerase chain reaction (RT-PCR) for miRNA profiling.
• Created reference samples using two methods: mixing extracted total RNA and mixing intact cells before RNA extraction.
• Developed in silico models to predict miRNA abundance ratios for target values.

Main Results:

• Successfully created and validated cell-based reference samples using both RNA mixtures and intact cell mixtures.
• MicroRNA profiling of the reference samples accurately replicated predicted target values.
• Both reference sample types demonstrated utility for inter- and intra-laboratory testing and mimicry of clinical specimens.

Conclusions:

• Cell-based reference samples can be effectively created for performance assessment from biomolecule extraction to quantitation.
• The demonstrated paradigm is adaptable to genome-scale platforms and other biomolecular endpoints beyond miRNA profiling.
• These reference samples provide a robust method for quality control, process monitoring, and validation in laboratory settings.