Importance of sample quality for genomics analysis, ethics & biobanking Flashcards
What legal and ethical matters must be considered?
- Declaration of Helsinki: Ethical principles for medical research involving human subjects, including research on identifiable human material and data.
- Health Research Authority: Established to promote and protect the interests of patients, streamline regulation and promote transparency in health and social care research.
- Human Tissue Act (2004): Regulates activities concerning the removal, storage, use and disposal of human tissue. Consent is the fundamental principle of the legislation.
- Human Tissue Authority: Regulates organisations that remove, store and use human tissue for research, medical treatment, post-mortem examination, education and training, and display in public.
Why was the human tissue act 2004 introduced?
To reassure the public that use of human cells and tissues are being used appropriately and with full knowledge and approval of the donor
Public outcry following discovery of organ retention for the purpose of ‘research’ at;
- Liverpool’s Alder Hey Children’s Hospital Bristol Royal Infirmary
- Manchester
without consent from patient, parent or next-of-kin
What are the human tissue authority standards?
Consent
Governance and quality control
- Standard operating procedures
- Risk Assessment
- Study protocol
- Laboratory manual
Premises, Facilities, Equipment
Traceability
- What is a legal requirement for the purpose of conducting research using human tissue since 1st Sept 2006?
- What are three key requirements?
Informed consent
This is an ongoing agreement by a person to receive treatment, undergo procedures or participate in research, after risks, benefits and alternatives have been adequately explained to them.
- Three key requirements:
– Consent should be given by somebody with the mental ability to do so
– Sufficient and appropriate information should be given to and understood by the participant
– Consent must be freely given
What is informed consent about? What does it apply to?
Information and building a relationship with the participant. Applies to both patients and healthy volunteers
What three steps are involved in attaining informed consent?
- Information
- Discussion
a. Mental capacity?
b. Inclusion criteria?
c. Understand research?
d. Understand what’s donated? - Agreement
a. Initialled
b. Signedbydonor
c. Signed by consent receiver
What is a research biobank?
Stored collections of biological samples linked to the donors personal information that can be used not just for one specific study but for different types of research undertaken by multiple investigators.
What are three types of biobanks?
Population Biobank: Consist of samples (often blood) and information on lifestyle and medical history from healthy volunteers.
Disease-specific Biobank: Consist of samples and information from individuals diagnosed with a similar type of disease, such as cancer or infectious diseases.
Organ specific Biobank: Consist of samples and information from individuals with a range of different diseases but in a specific organ, such as the benign brain disease bank or a liver biobank
What impacts blood sample DNA quality & quantity?
Time from blood draw to isolation/preservation of WBCs
Does time from blood draw to isolation/preservation of serum impacts on quality proteins, electrolytes, antibodies etc??
Yes and therefore serum must be processed rapidly
What is hypoxia?
Tissue lacks oxygen. Once tissue is removed hypoxia begins therefore must immediately be preserved.
If a tissue sample is attained from a surgical treatment, what is needed?
Pathology dissection
It is important to record aspects what?
Tissue handling (eg hypoxia times, fixatives)
How is frozen tissue attained and used?
- Fresh specimen - described, measured, inked, sliced
- Tissue cut into 5 x 5 x 10mm blocks, placed on card, coated in OCT, frozen using isopentane cooled in liquid nitrogen and placed in labelled cryotube.
• Freeze as quickly as possible to maximise preservation - 30min max
Use
• Frozen piece: DNA or RNA extraction
• Frozen section: Immunohistochemistry, Immunofluorescence, In situ hybridisation, needle macro-dissection or laser capture micro-dissection for DNA/RNA extraction.
How is Formalin-Fixed, Paraffin-Embedded Tissue attained and used?
- Specimen described, sliced and fixed in buffered formalin
- Tissue cut into slices 2-3mm thick and placed in cassette • Paraffin –wax processed (overnight)
Use
• 3m sections: Immunohistochemistry, Immunofluorescence, In situ hybridisation.
- > 10m sections: needle macro-dissection or laser capture micro-dissection for DNA/RNA extraction
- Tissue Microarray (TMA) construction
