BPV Analytical Platforms
Highly annotated biospecimens were collected for the Biospecimen Pre-analytical Variables (BPV) program through an organized and well-controlled infrastructure of Biospecimen Source Sites (BSS). Various analytical platforms were used by BSS to determine pre-analytical variable effects on a wide range of studies including DNA methylation, copy number variation, genotyping, mRNA expression, miRNA expression and protein expression.
BSS receive in-person training on the proper use of standard operating procedures developed by NCI to ensure that uniform collection and handling procedures are used across all sites. All BPV biospecimens are collected using kits provided by the Comprehensive Biospecimen Resource (CBR), a centralized site that assembles collection and shipping kits, receives most biospecimens from the BSSs for processing and storage, and ships biospecimens from the BSSs to the processing and analysis facilities.
BSS use a web-based interface (the Comprehensive Data Resource- CDR) to enter data regarding collection, processing, and handling of donor biospecimens and clinical information as well as pathological evaluation of the tumor. Over 300 common data elements are collected in CDR for each case, which are reviewed by data quality managers. BPV specimens are tracked via specimen IDs and unique barcodes using the Laboratory Information Management System (LIMS).
Figure 1. Analysis, cancer type and platforms used in the BPV program
Study | Cancer Type | Platform and Analysis |
---|---|---|
DNA Methylation | Kidney | MethyLight- methylation-specific PCR amplification of methylation markers |
Copy Number Variation | Kidney | Genome e-wide CNV evaluation by array-based genomic hybridization (aCGH) |
Genotyping | Kidney, Colon, Ovary | Whole exome sequencing by NGS |
RNA Expression | Kidney, Colon, Ovary | Transcriptome profiling by NGS |
miRNA Expression | Kidney, Ovary | Human miRNA profiling by WaferGen SmartChip (Human miRNA Panel) |
miRNA Expression | Kidney | Oncopanel evaluation by TaqMan Open Array |
miRNA Expression | Kidney | Cold-ischemic biomarker expression by TaqMan |
miRNA Expression | Kidney, Colon, Ovary | Expression biomarkers by RNA in-situ hybridization (RNAscope) |
Protein Expression (Tissue) | Kidney, Colon, Ovary | Cancer specific biomarker expression by Immunohistochemistry (IHC) assay |
Protein Expression (Tissue) | Kidney, Colon, Ovary | Total protein and phosphoprotein expression by MassSpec |
Protein Expression (Plasma) | Kidney, Colon, Ovary, Lung | Targeted protein expression by multiplexed Multiple Reaction Monitoring (MRM) assay |
Protein Expression (Plasma) | Kidney, Colon, Ovary, Lung | Cancer specific protein expression by Luminex assay (HumanMap) |
Metabolite Profile (Plasma) | Kidney, Colon, Ovary, Lung | Global metabolite profiling by MassSpec |