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Bone Marrow Aspirate

Written by Mark Hill on .

Bone Marrow aspirates are processed in the Immunophenotyping laboratory. Slides are routinely stained with May-Grunwald/Giemsa and also a Perls Prussian Blue stain for iron stores. Aspirate samples should be delivered to the laboratory as soon as possible for staining as immunophenotyping may also be required on the same bone marrow samples following microscopy. Trephine samples should be directed to the Histopathology Department with an accompanying Histopathology request form.

 

The Bone Marow Aspirate Request Form (pdf) can be downloaded from here.

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Haemoglobinopathy Screening

Written by Mark Hill on .

Screening for thalassaemia and variant haemoglobins (such as Sickle haemoglobin) is performed within the Haematology laboratory at Heartlands Hospital for all Trust sites and GP's. Patients may require haemoglobinopathy testing as part of ante-natal screening or pre-operative assessment.

Requests should be made on a Haematology request form and sent with a 4ml EDTA sample for analysis. We are unable to reflex haemoglobinopathy screening on suspected full blood counts results without a request. Ante-natal requests should be accompanied with a Family Origin Questionnaire.

Occasionally samples are referred to Central Middlesex Hospital, London or John Radcliffe Hospital, Oxford for confirmation of new or unusual variants or for alpha thalassaemia testing.

For further information or advice please contact the laboratory on extension 40198 or check the Test Database.

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Immunophenotyping (Haematology)

Written by Craig Webster on .

We offer an extensive repertoire of investigations for acute leukaemia, lymphoproliferative disorders, stem cells (CD34), PNH and also for the quantitation of feto-maternal haemorrhage. For haematological malignancies, a consultant will examine the morphology and comment on the immunophenotyping results. Preliminary results are available within 24 hours of receipt of the sample and a final report sent in the post following Consultant review.

Immunophenotyping requires fresh blood or bone marrow drawn into EDTA tubes. The laboratory is open Monday - Friday, 08:30 - 17:30. Latest time of sample receipt for same day analysis is 2.00pm. Samples arriving later than this will be refrigerated until the next working day. Analysis may be performed following weekend storage providing cell viability is within acceptable limits.

Samples should be transported to the laboratory as soon as possible after collection. For external requests, samples should be transported in suitable containers via courier and labelled for the attention of Cell Markers, Haematology BHH. Samples should not be sent in the post.

For enquiries telephone 0121 424 0704 to speak with a Senior Biomedical Scientist in the Immunophenotyping Laboratory. For clinical advice please contact the on-call Consultant Haematologist via switchboard 0121 424 2000.

Click here to download a request form (pdf). Please complete request form accurately providing as much relevant information as possible. The laboratory will not accept samples/requests with missing or mislabelled PID.

Click for Test Database

Additional Information for Immunophenotyping

Investigation of Acute Leukaemia

An extensive 10 colour panel and morphology review in order to provide a diagnosis for acute leukaemia. The laboratory may telephone interim results prior to Consultant review. Our aim is to provide a same day turnaround dependant on time of arrival of sample. Please telephone in adavance of all suspected acute leukaemia cases.

MDS / Acute leukaemia screen

A quick screen based on the acute leukaemia panel in order to identify blast populations and basic phenotype of cells.

Investigation of lymphoproliferative disorder

An extensive panel to diagnose chronic lymphoproliferative disorders such as chronic lymphocytic leukaemia and lymphoma. A Matutes score will be reported and ZAP70 to aid prognosis. Please note ZAP70 will only be performed on samples less than 48 hours old and with a lymphocyte count greater than 5x10*9/L. 

Lymphocytosis / CLL screen

A quick screen to identify clonal populations of lymphocytes. A full lymphoproliferative screen will follow if appropriate.

CLL MRD

A simple screen for known CLL patients to identify populations of clonal lymphocytes.

Hairy cell leukaemia panel

A lymphoproliferative panel designed specifically for cases suspected of hairy cell leukaemia.

Myeloma / MGUS Panel

A six colour panel including CD38 and CD138 to identify plasma cell populations to aid in the diagnosis of myeloma and monoclonal gammopathy of undetermined significance (MGUS).

PNH Screen

A screen using both red blood cells and white blood cells (granulocytes) to identify PNH clones. Screening of wbc's incorportaes the use of FLAER. Please note that bone marrow is unsuitable for PNH testing due to the presence of immature cells and recently transfused patients may give reduced or negative red cell populations in those affected.

Click here for PNH request guidance

Feto-maternal haemorrhage

To identify the presence of and enumerate the bleed of foetal red blood cells into the maternal circulation. Patients from HEFT will automatically have FMH samples analysed by flow cytometry if the bleed is calculated as high by the Kleihauer method (in Blood Bank). Please note that this screen is based on the principle that maternal samples are Rh D negative and baby/cord samples are Rh D positive. In ante-natal patients or patients who have had an IUD it may not be possible to identify a bleed if mom and baby have the same Rh D type. Also as the principle is based on staining using a flourescent anti-D, patients who have received recent prophylactic anti-D may give reduced values due to antigen blocking by prophylactic anti-D.

For any further advice please telephone the laboratory on 0121 424 0704  

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Molecular Haematology

Written by Craig Webster on .

Many diseases are known to be caused by mutations in a persons DNA. The polymerase chain reaction (PCR) is a technique used to amplify specific regions of DNA for analysis. Using PCR we can target those specific regions to identify if a mutation is present or not. This allows us to make a diagnosis or prognosis of a disease or disorder.

The Molecular Haematology laboratory offers investigations for haematological malignancy and molecular carrier status of various haematological disorders. Below is a brief explanation of the tests on offer.

For details regarding sample requirement please see the Test Database. Results are usually available within 10 working days of sample receipt.

Lymphoproliferative Disorders

IgH – The diagnosis of B-cell lymphoid malignancies can be substantiated by detecting clonal immunoglobulin heavy chain gene rearrangements. The Department employs polymerase chain reaction (PCR) techniques for the framework 3 region (FR3) of the IgH gene. It should be noted that the sensitivity of this technique using blood or bone marrow aspirates is in the region of 72%.

TCR – The diagnosis of T-cell lymphoid malignancies can be supported, although not diagnostic, by detecting clonal populations of T-cell gene rearrangements. The PCR amplifies across the rearranged TCRγ gene found in all T cells and is based on two separate reactions; the first uses a primer directed against the consensus, γC, region of the variable gene, the second directed against γ9.

Thrombophilia Disorders

Factor V Leiden (R506Q) - Activated protein C resistance has been attributed to a single point mutation in the gene that codes for the clotting factor protein Factor V, and termed R506Q-Factor V Leiden. This polymorphism involves a single base pair substitution at position 1691 in the factor V gene, resulting in substitution of the arginine (R) at position 506 by glutamine (Q). The mutation results in an increased risk of thrombosis. Exposure to certain environmental risk factors, such as surgery, pregnancy, or oral contraceptives, can further increase the risk for a venous thrombotic event.
 
Prothrombin Gene (G20210A) - Prothrombin is the precursor to thrombin, which plays a central role in thrombosis, haemostasis and fibrinolysis. A mutation in the prothrombin gene, a single G to A transition at position 20210, causes an increase in prothrombin and is associated with an increased risk of venous thrombo-embolism.
 
MTHFR (C677T) - MTHFR is a regulatory enzyme in homocysteine metabolism. A C677T mutation in the gene has been identified and homozygosity for the mutation has been implicated in mild hyperhomocysteinaemia. Hyperhomocysteinaemia results in a 2.5 fold increased risk for venous thrombo-embolic events.

Haemochromatosis Screening

C282Y, H63D - Haemochromatosis is an autosomal recessive disorder of iron overload. The disease is characterised by the progressive accumulation of dietary iron, which can lead to clinical complications including cirrhosis, diabetes, cardiomyopathy, arthritis and hepatocellular carcinoma. Several point mutations have been identified, however two are considered to be of more importance in the disease, C282Y and H63D. 80 – 100% of patients with Hereditary Haemochromatosis are homozygous for the C282Y mutation, and individuals showing compound heterozygosity may also be predisposed to iron overload.
 
DNA retrieved from samples is currently stored for up to 30 years. Any additional request on an archived sample should be made on a Molecular request form for up to this time period. The quality of the DNA will determine if the request is achievable.
 
The Molecular Haematology Laboratory is staffed Monday to Friday 0900 to 1730. Any samples arriving outside of these hours will be refrigerated overnight and dealt with the next working day. For enquiries telephone 0121 424 3704 to speak to a member of Molecular Haematology staff.
 
Click here to download a request form (pdf). Please complete request form accurately providing as much relevant information as possible. The laboratory will not accept samples/requests with missing or mislabelled PID.
 
Further information can be obtained from the following external websites:
 
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Referred Tests

Written by Craig Webster on .

Referred Tests

The following referred tests should be requested on a Haematology request form and the Department will make arrangements to send the sample on.

Abnormal Haemoglobinopathies

The Department offers a comprehensive haemoglobinopathy screening service, however occasionally samples are referred for further analysis or confirmatory testing. Laboratory staff will refer samples if appropriate.

Unidentified haemoglobin variants

Haematology Department
Haemoglobinopathy Laboratory
Central Middlesex Hospital
Acton Lane
London
NW10 7NS
 
or
 
High risk alpha thalassaemia
 
Molecular Haematology Department
John Radcliffe Hospital
Headley Way
Headington
Oxford
OX3 9DU
Ristocetin Co Factor

Ricof samples are referred to Queen Elizabeth Hospital, Birmingham. Please discuss with a Consultant Haematologist prior to requesting this test. 

Special Coagulation Laboratory
Haematology Department
Queen Elizabeth Hospital
Edgbaston
Birmingham

Pyruvate Kinase

Pyruvate Kinase assays are referred to Hammersmith Hospital. Please discuss with a Consultant Haematologist prior to requesting this test.

Special Haematology
Haematology Department
Hammersmith Hospital
Du Cane Road
London
W12 0HS

Positive Malaria Results

When a positive malaria result is obtained we are required to refer blood films to the Malaria Reference Laboratory.

London School of Hygiene and Tropical Medicine
Keppel Street
London
WC1E 7HT

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