Genetically characterized positive control cell lines derived from residual clinical blood samples

Area of Science:

• Molecular Biology
• Genetics
• Clinical Diagnostics

Background:

• Positive control materials for molecular genetic testing are scarce.
• Epstein-Barr virus (EBV)-transformed peripheral blood lymphocyte cell lines offer a source of high-quality DNA.
• A process was developed to generate and validate these cell lines from residual patient samples.

Purpose of the Study:

• To develop a reliable process for producing and validating positive control cell lines.
• To address the critical shortage of positive control materials in clinical molecular genetic testing.
• To establish stable cell lines with clinically relevant mutations for diagnostic applications.

Main Methods:

• A network of 32 genetic testing laboratories collected residual blood samples with known mutations.
• Peripheral blood lymphocytes were transformed with Epstein-Barr virus (EBV).
• Established cell lines were validated for specific mutations and stability in culture.

Main Results:

• 113 residual blood samples with mutations for 14 genetic disorders were collected.
• 41 EBV-transformed cell lines were successfully established.
• 33 cell lines were validated as positive controls for clinical diagnostic applications, with mutations remaining stable.

Conclusions:

• A robust process for generating and validating positive control cell lines from residual clinical samples has been established.
• This process can help alleviate the ongoing shortage of essential positive control materials for molecular genetic testing.
• The validated cell lines are suitable for use in clinical diagnostic settings.