Sample requirements

Tube types

The types of tubes that should be used for blood samples are noted in the detail above. In general there are two types, EDTA and LitHep and it is important the correct one is used as stated.

EDTA (or ethylenediaminetetraacetic acid) is most often used for haematological tests (red and white blood cell count, Hb, blood group). It functions by binding calcium in the blood and keeping the blood from clotting.

• What these tubes look like: EDTA tubes tend to have purple tops though there are a variety of brands available, so they can have other colour tops (e.g. red or blue).

LitHep tubes are coated with Lithium Heparin on the inside. It is used for is used for clinical chemistry tests such as cholesterol, CRP, hormones etc. It interferes with PCR, so cannot be used for tests that require this.

• What these tubes look like: LiHep tubes usually have orange or green tops, though again some brands may use different colour tops.

Both tube types are very widespread and any phlebotomist should have them.

Please note:

The laboratory is unable to accept blood samples or saliva samples from patients who have received an allogenic haematopoietic stem cell transplant (including bone marrow transplant) due to the presence of donor DNA. In these patients, a buccal swab sample or a skin biopsy should be referred. Please contact the laboratory for further advice on sample requirements.

Peripheral blood samples

The default choice for blood tubes is EDTA for the majority of tests in the Rare and Inherited Disease Test Directory.

This includes DNA investigations.

Lithium Heparin blood tubes are ONLY required for the following tests:

• R402.1 Premature ovarian insufficiency - karyotype
• R314.2 Ambiguous genitalia presenting neonatally - karyotype
• R297.1 Possible structural chromosomal rearrangement - karyotype
• R298.1 Possible structural or mosaic chromosomal abnormality - FISH
• R265.1 Chromosomal mosaicism - karyotype

Plus the following DNA repair defect tests:

• R227.2 Xeroderma pigmentosum, Trichothiodystrophy or Cockayne syndrome
• R258.1 Cytopenia - Fanconi breakage testing indicated
• R259.1 Nijmegen breakage syndrome
• R260 Fanconi anaemia or Bloom syndrome - chromosome breakage testing
• R294.1 Ataxia telangiectasia - DNA repair testing

Volumes required:

Adults: 5ml blood
Children: 1 to 2ml blood

Saliva, mouthwash or buccal swab samples

The laboratory will accept saliva, mouthwash or buccal swab samples for certain genetic tests. Please contact the laboratory prior to arranging the sample.

Due to risks associated with respiratory illness, we only accept samples collected in GenoTek Oragene mouthwash/saliva kits and GenoTek Oragene Buccal Swab kits. Samples received in alternative collection kits will be discarded.

Skin sample

Please contact the laboratory prior to arranging a sample.

4mm2 (minimum 3mm2) sample in sterile tissue transport medium (provided by the laboratory) within leak-proof plastic container.

Cultured cells

Cultured cells in sterile container or fixed cell suspension in a leak proof container.

DNA samples

Samples of extracted DNA (50ul DNA at concentration of ~500ng/ul) should be in a leak proof plastic container and sent at room temperature.

Sample for RNA analysis

Please contact the laboratory prior to arranging sample

Other sample types

Please contact the laboratory to discuss requirements prior to arranging the sample.

Please contact the laboratory prior to arranging a prenatal sample using our pre-natal pre-notification form.

Amniotic fluid samples

10 to 20ml in sterile leak-proof container.

Chorionic villi and Placental membranes samples

Chorionic villi - 10-30mg of tissue in sterile tissue transport medium within leak-proof plastic container.

Placental membranes in sterile tissue transport medium within leak-proof plastic container.

Contact the laboratory for information on appropriate tissue transport medium.

Products of conception samples

Place in a sterile leak-proof container.

Skin biopsy

4mm2 (minimum 3mm2) sample in sterile tissue transport medium (provided by the laboratory) within leak-proof plastic container.

Non-invasive prenatal diagnosis testing

Please contact the laboratory prior to arranging sample.

For Next Generation Sequencing, please send 2 H&E stained slides and 8 unstained slides with the tumour area marked clearly on the stained slide and the % tumour assessment clearly stated on the request form.

All slides for molecular testing should be mounted on charged slides

Slides must be 10x 4µm thick sections (multiple sections per slide, as will fit). H&E stain sections 1 and 10

Unstained slides should be dried at 37oC for a minimum of 1 hour, or up to overnight before being transported to the GLH

For FISH testing – please cut 2µm thick sections (maximum 2x separated sections per slide). H&E stain section 1

For any other tests please send 2 unstained 4µm slides per test

Note: For CUH only

Bone Marrow

Cytogenetic analysis

Bone marrow aspirate (1-2mL) in bone marrow transport medium containing RPMI medium, heparin and antibiotics (available from the laboratory on request). If unavailable, samples should be placed in lithium heparin.

Samples for CLL and Multiple Myeloma FISH should be sent in EDTA.

Molecular genomic analysis

A separate peripheral blood (EDTA) or bone marrow (EDTA) sample is required (usually 5-10 mL; 20 mL peripheral blood for BCR-ABL1 quantification and post-transplant chimerism analysis). To ensure RNA integrity samples should be received within 24 hours of collection. For monitoring of residual disease by RT-PCR, samples should be received in the laboratory on the day of collection. Aged samples (>48 hours) may be rejected, especially if BCR-ABL1 quantification is requested, as tests may not be reliable in such cases.

T-cell and B-cell Clonality sample requirements

Depending upon the type of biopsy there are differential sample requirements which are outlined in a specific guidance document (below). Please note that two H&E stained slides (first and last slide) are required for all cases and the lesional area should be marked on the H&E slides. A copy of the histopathology report should also be provided as this will aid in the molecular interpretation of these cases.