Molecular Protocol Development & Validation

Protocols for molecular analysis of biospecimens are developed in a similar manner as histology protocols – appropriate negative and positive controls for a given molecular target are first identified and used to develop the experimental conditions. These control materials can be samples with known mutations, well characterized cell lines, or commercially available samples.

Once the experimental conditions are determined, the protocol is validated on human samples and the data is interpreted by a pathologist who is board certified in molecular genetic pathology. Protocols are validated in order to determine the dynamic range of the assay and the limit of detection. Following the validation, the protocol is written and reviewed before it becomes standard operating procedure.

Any subsequent modifications to the protocol are validated using the appropriate materials before these modifications become incorporated to the standard operating procedure. Once a protocol is established and validated, QC/QA is maintained by routinely testing known samples to determine reproducibility and by including appropriate positive and negative controls on every experiment.

Following the development and validation of a specific protocol, the CTRL is positioned to perform testing of human specimens. These specimens can be archival samples collected during routine patient care and stored in the pathology department. Alternatively, these samples can be prospective studies collected specifically for a research project and/or obtained during routine patient care. In either case, all samples are initially reviewed by a member of the pathology department in order to render a diagnosis.

Samples destined for subsequent research projects are analyzed for adequacy – this includes total cellularity, relative tumor cellularity, sample size, sample quality, DNA/RNA yields, and DNA/RNA quality. Appropriate samples are then available to use for future research studies, whereas samples determined to be inadequate for any reason are not included in any subsequent research activities.