Molecular Core Lab
MCL: Research Platforms and Projects
Introduction
One of the primary missions of the Surgical Critical Care Initiative (SC2i) is to advance the bench to bedside approach of connecting translational research to patients and patient care. The Molecular Core Laboratory (MCL) of the SC2i serves as one of the primary research labs of the organization that help design and execute multi-omics based assays and biobanking. The College of American Pathologists (CAP) consecutively recognized the MCL’s biorepository in The CAP Accreditation Program, which is widely recognized as the gold standard and serves as a model for various federal, state, and private laboratory accreditation programs throughout the world. Only 73 biorepositories in the United States are currently accredited. The SC2i MCL, in coordination with the Clinical Research Team and Regulatory and Laboratory Compliance Team, has identified close to 400 biorepository requirements. Additionally, we continually improve existing documents, such as the Quality Management Plan, Standard Operating Procedures, guidance documents, and training manuals to uphold our current and future laboratory accreditations.
The MCL of SC2i, which is part of the Uniformed Services University of the Health Services and the Walter Reed National Military Medical Center, received its initial biorepository accreditation in February 2020. Receiving and maintaining CAP accreditation is an exciting milestone in accomplishing one of the SC2i missions in developing biomarker-driven Clinical Decision Support Tools (CDSTs). The next goal for SC2i is to obtain FDA approval for one of our CDSTs, which requires ongoing approval in the CAP Laboratory Accreditation Program and Clinical Laboratory Improvement Program (CLIP) certification of their laboratories for high complexity clinical testing.
Platforms
- MesoScale Discovery (MSD):
Meso Scale Discovery (MSD) assay is a 96-well bioanalysis immunoassay-based platform (ELISA) that utilizes electrochemiluminescence for multiplexing. Multiplexing is a technique that allows multiple analytes to be quantitated within a single well. SC2i has worked diligently to expand the detection range for 56 immunological-related analytes. These analytes have been reported to play critical roles in angiogenesis, inflammation, immune cell activation/suppression, chemotaxis, and endothelial/epithelial homeostasis. SC2i continues to enhance the identification of potential prognostic and diagnostic biomarkers that may be present in various biospecimens (serum, wound effluent, peritoneal fluid vs lavage) to aid in the development of future CDSTs.
- B·R·A·H·M·S KRYPTOR:
B·R·A·H·M·S KRYPTOR is a fully automatic, closed laboratory analysis system that can perform numerous various analyses in random-access operations using a unique measuring principle called TRACE (Time Resolved Amplified Cryptate Emission). Precision is vital for the clinical decision-making process that is supported by the measured result of a biomarker. This requires both a stable and well-defined biomarker assay on a platform that safeguards a reliable and precise measurement of the biomarker. Precision technology paves the way for the development of innovative in-vitro diagnostic reagents for diverse clinical areas. SC2i is currently expanding the range of assays designed to run on our B·R·A·H·M·S KRYPTOR Analyzer. These assays include: 1) Procalcitonin (PCT)(21 CFR 866.3215, Class II) – identify bacterial infections and development of sepsis 2) Copeptin (AVP/ADH) – surrogate marker for vasopressin to help monitor disease progression and critically ill patients.
- qRT-PCR and Nanostring:
Quantitative real-time polymerase chain reaction (qRT-PCR) is a well-established method for the amplification, detection, and quantification of mRNA and miRNA gene expression in the areas of pre-clinical and clinical diagnostics. MCL currently utilizes this scientific platform to help identify and characterize the expression levels of existing as well as potentially novel biomarkers that may be predictive or diagnostic of patient care.
Nanostring is novel and dynamic new technology within the field of spatial biology and is 21 CFR Part 11 compliant. Nanostring provides a cost-effective automated solution for multiplex analysis of 800+ gene targets within a simple yet highly reproducible workflow. Gene expression can be quantified and analyzed using their ready-to-go or custom panels for a multitude of nucleic acid-based sample types (Total RNA, cell lysates in GTIC, FFPE-derived total RNA and PAXgene-lysed whole blood) and research fields (immunology, oncology, infectious disease, etc). These processes help identify unique gene signatures and cellular pathways that can be utilized to make better and more informed decisions regarding patient care.
- Bioprocessing:
SC2i’s CAP accredited Biorepository include a vast array of patient samples (civilian and DoD) across multiple clinical cohorts and timepoints that link to the mission of SC2i. In coordination with our consortium partners, we collect patient samples from different geographical and clinical populations that can be utilized in our multi-omic scientific research investigations to facilitate the identification and development of biomarker-based CDSTs. The laboratory processing of the samples aims to collect and store an assortment of different specimen types that can be used across our multiple investigative and analytical platforms downstream. The processing includes, but is not limited to, collecting and cryopreservation of whole blood, serum, urine, wound effluent, isolation of peripheral blood mononuclear cells (PBMC), and wound tissue. These samples are collected, processed, persevered, and stored to the highest standard so quality data can be obtained at later by any of our participating entities. SC2i’s mission is to uphold the highest standard of biobanking practices to allow these unique clinical events to be studied to further the insight into predictive and diagnostic patient care, as well as improve overall patient outcome.