The pathology Service of the Western Sussex Hospitals NHS ...



Haematolgy Tests

Full Blood Count (FBC):

Abbott Cell Dyn Sapphire analysers produce a complete automated blood count result with a differential and platelet count - individual parameters need not be requested. Reticulocytes and Nucleated Red Blood Cell Analysis can also be undertaken where appropriate.

Blood films are made and examined if requested or if the parameters suggest that a manual film and/or differential will be helpful.

Erythrocyte Sedimentation Rate (ESR) :

The ESR is an indirect measure of the degree of inflammation present in the body. It is a non-specific test and has to be interpreted within the clinical context in which it is requested. It may be helpful in diagnosing inflammatory disorders such as temporal arteritis/polymyalgia rheumatica and may also be used to monitor disease activity and response to therapy in these and other inflammatory disorders. It is affected by age, gender and anaemia.

Analysed on request.

Paul Bunnell Test:

A test for the presence of heterophile antibodies in the serum produced in infectious mononucleosis.

Malarial Parasites: Thick and thin blood films are stained with Giemsa stain and examined for the presence of malarial parasites. A rapid immunological test is also performed as a complementary test to the standard blood film examination.

Blood films for malarial parasites will be made on request if clinical indications suggest the patient is at risk of malaria. Positive blood films are sent to the London School of Hygiene and Tropical Medicine for confirmation.

SAMPLE SHOULD ARRIVE IN THE LABORATORY NO LATER THAN 4 HOURS AFTER BEING TAKEN.

Haemoglobinopathy Screening:

Haemoglobinopathy screens can be requested for any patient who might be considered to be at riskof having a haemoglobin variant or thalassaemia. A haemoglobinopathy screen consists of a Full Blood Count and Hb HPLC analysis. If haemoglobin variants are detected by HPLC, confirmation will be performed by electrophoresis and the Sickle Solubility Test.Rare haemoglobin variants that cannot be identified by our laboratory are referred to the red cell reference centre at Central Middlesex Hospital.

These tests should be requested by the patient’s clinician with prior consent. The laboratory does not take telephone requests for add-on haemoglobinopathy screening unless made by a ConsultantHaematologist.

Sickle Cell Solubility Screen:

This is a rapid test to detect the presence of sickle haemoglobin, eg. HbS but in itself doesn’t

distinguish between sickle cell trait and sickle cell disease. Positive screen results will be referred for further confirmatory tests. This test is performed when Hb HPLC indicates the presence of a haemoglobin variant or if a rapid result on sickle status is required.

Antenatal Sickle Cell and Thalassaemia screening:

WSHFT participates in the National Screening Programme as a “low prevalence area”. Screening is performed according to the National Screening Programme guidelines using FBC, family of origin questionnaire and, when indicated, HPLC.

Glucose-6-phosphate dehydrogenase (G-6-PD) Screening Test:

G-6-PD deficiency is the most common inherited genetic enzyme deficiency. Most individuals are

asymptomatic, but devastating haemolysis can occur when susceptible patients are exposed tooxidative drugs or infection. It is important to identify individuals at risk as certain drugs may then beavoided. The screening test is based on a qualitative visual fluorescence screening procedure.

Cerebrospinal Fluid (CSF) Analysis

CSF is centrifuged, the resulting preparation stained and a manual count made of any cells present.

Usually only performed if requested by Haematology Registrar/Consultant if there is a suspicion of CNS involvement in haematological neoplasms. Any other reasons for cell counting should be referred to Microbiology or Cytology.

Sample requirement: 1ml CSF.

Bone Marrow Sample Analysis:

Bone Marrow samples are only taken by a Haematology medical staff.

All diagnostic samples (morphology, cytogenetics, molecular genetics) are sent to Kings College Hospital (HMDC) for analysis. Depending on the clinical indication some bone marrow aspirate slides are also processed in duplicate at WSHFT to allow for a provisional result and assist with patient management.

Turn-around times depend on the assays required. Discuss with haematologist.

Peripheral blood Leucocyte Immunophenotyping:

Samples are sent to Kings College Hospital HMDC. See above

Sample Requirements/ Turnaround Times/Other Information

|TESTS |SPECIMEN |TURNAROUND |SPECIAL |KEY FACTORS |

| |REQUIRED |TIMES |INSTRUCTIONS |AFFECTING TEST PERFORMANCE OR |

| | | | |INTERPRETATION |

|Full Blood Count |4ml Mauve top EDTA or |4 hrs | |Red cell Cold autoagglutinns Lipaemia |

|(FBC) |1.4 Mauve Paediatric |1 Hour for urgent | |(these are corrected in lab) |

| |top |samples | | |

|Erythrocyte |4ml Mauve top EDTA |6 hrs |Minimum volume 2ml |Red cell Cold autoagglutinns (test will|

|Sedimentation Rate | | | |not be reported) |

|(ESR) | | | | |

|Reticulocyte count |4ml Mauve top EDTA |4 hrs | | |

|Blood film (manual |4ml Mauve top EDTA |24 hrs where clinically | |Prolonged exposure to EDTA |

|differential) | |Urgent otherwise upto 72| |anticoagulant may result in abnormal |

| | |hours. | |cell morphology. |

|Infectious |4ml Mauve top EDTA |8 hrs | | |

|mononucleosis screen (Glandular| | | |Some patients do not |

|fever) | | | |produce antibody. |

| | | | |In early stage antibody may be |

| | | | |undetectable. |

|Malarial Parasites |4ml Mauve top EDTA |4 hrs |Rapid test within |If initial screen negative this should |

| | | |1 hr. |be repeated if strong suspicion of |

| | | | |malaria present. Ensure been to VHF |

| | | | |regiosn VHF questionnaire sent with |

| | | | |sample |

|Haemoglobinopathy |4ml Mauve top EDTA |3 days |Urgent requests |Requestor should indicate if patient |

|Screen. | | |will be processed |has been recently transfused |

| | | | | |

|Hb S Screen | | | | |

|Glucose-6-phosphate |4ml Mauve top EDTA |3 days | |Raised reticulocyte count |

|dehydrogenase | | | |may result in false normal result |

|(G6PD) | | | | |

|Bone marrow aspirate |Bone marrow |14 days |Discuss with |Refrigerate if not sending straight to |

| | | |Haematologist |laboratory. Prolonged exposure to EDTA |

| | | | |anticoagulant may result in abnormal |

| | | | |cell morphology. |

|Bone marrow trephine biopsy |Bone marrow |14 days |Discuss with | |

| | | |Haematologist | |

|CSF Cytospin |CSF fluid in plain | |Send sample to laboratory|Haematology patients only |

| |bottle | |before 16:30 | |

Reference Ranges

Full Blood Count Parameters and Cell Counts

Reference Ranges for Infants

|Test |Birth |Day 3 |Day 7 |Day 14 |1 month |2 months |3-6 months |

|Hb g/l |140 - 220 |150 – 210 |135 - 215 |125 - 205 |115 - 165 |94 - 130 |111 - 141 |

|Hct l/l |0.450–0.750 |0.450 – 0.670 |0.420–0.660 |0.310 – 0.710 |0.330 – 0.530 |0.280 – 0.420 |0.300 – 0.400 |

|MCV fl |100 - 120 |92 - 118 |88 - 126 |86 - 124 |92 - 116 |87 - 103 |68 – 84 |

|MCHC g/l |300 - 360 |290 - 370 |280 - 380 |280 - 380 |290 - 370 |285 - 355 |300 - 360 |

|Retic x109/l |120 - 400 |50 - 350 |50 - 100 |50 - 100 |20 - 60 |30 - 50 |40 - 100 |

|Neuts x109/l |4.0 – 14.0 |3.0 – 5.0 |3.0 – 6.0 |3.0 – 7.0 |3.0 – 9.0 |1.0 – 5.0 |1.0 – 6.0 |

|Mono x109/l |0.5 – 2.0 |0.5 – 1.0 |0.1 – 1.7 |0.1 – 1.7 |0.3 – 1.0 |0.4 – 1.2 |0.2 – 1.2 |

|Plats x109/l |100 - 450 |210 - 500 |160 - 500 |170 - 550 |200 - 500 |210 - 650 |200 - 550 |

Reference: Dacie & Lewis Practical Haematology 12th Edition

Reference Ranges from Children to Adults

|Test |1 Year |2-6 Years |6-12 Years |Adult male |Adult female |

|Hb g/l |111 – 141 |110 - 140 |115 – 155 |130 - 170 |120 - 150 |

|Hct l/l |0.300 – 0.380 |0.340 – 0.400 |0.350 – 0.450 |0.400 – 0.500 |0.360 – 0.460 |

|MCV fl |72 - 84 |75 - 87 |77 – 95 |83 - 101 |83 - 101 |

|MCH pg |25.0 – 29.0 |24.0 – 30.0 |25.0 – 33.0 |27.0 – 32.0 |27.0 – 32.0 |

|MCHC g/l |320 - 360 |310 – 370 |310 – 370 |315 - 345 |315 - 345 |

|Retic x109/l |30 - 100 |30 – 100 |30 – 100 |50 - 100 |50 - 100 |

|WBC x109/l |6.0 – 16.0 |5.0 – 15.0 |5.0 – 13.0 |4.0 – 10.0 |4.0 – 10.0 |

|Neuts x109/l |1.0 – 7.0 |1.5 – 8.0 |2.0 – 8.0 |2.0 – 7.0 |2.0 - 7.0 |

|Lymphs x109/l |3.5 – 11.0 |6.0 – 9.0 |1.0 – 5.0 |1.0 – 3.0 |1.0 – 3.0 |

|Mono x109/l |0.2 – 1.0 |0.2 – 1.0 |0.2 – 1.0 |0.2 – 1.0 |0.2 – 1.0 |

|Eos x109/l |0.1 – 1.0 |0.1 – 1.0 |0.1 – 1.0 |0.02 – 0.5 |0.02- 0.5 |

|Baso x109/l |0.02 – 0.1 |0.02 – 0.1 |0.02 – 0.1 |0.02 – 0.1 |0.02 – 0.1 |

|Plats x109/l |200 - 550 |200 - 490 |170 - 450 |150 - 410 |150 - 410 |

Reference: Dacie & Lewis Practical Haematology 12th Edition

ESR Reference Ranges for adults

|Test |17-50 Years |50-61 Years |61-70 Years |>70 Years |

|ESR (male) mm/hr |≤ 10 |≤ 12 |≤ 14 |≤ 30 |

|ESR (female) mm/hr |≤ 12 |≤ 19 |≤ 20 |≤ 35 |

Reference: Dacie & Lewis Practical Haematology 12th Edition

HbA2 and HbF Reference Ranges for adults

|Test | |

|HbA2 % |2.2 – 3.5% |

|HbF % |< 1.0% |

Reference: Dacie & Lewis Practical Haematology 12th Edition

Routine Coagulation Tests

Coagulation Screening:

Prothrombin Time/International Normalised Ratio (PT/INR) and Activated Partial Thromboplastin Time (APTT) are our standard coagulation tests. Prolonged clotting times are an indication of abnormal clotting which could lead to an increased risk of bleeding. Further coagulation investigations may be reflexed by the laboratory depending on the nature of the coagulation results and previous test results to aid interpretation.

Clauss fibrinogen is reflex tested if the coagulation analyser indicates that the fibrinogen level is likely to be low (derived fibrinogen). Fibrinogen can also be requested by the requesting clinician.

When requesting coagulation screening tests it is important to include relevant clinical information as to why the test was requested to enable laboratory staff to determine the best course of action inthe event of an abnormal result.

It is not appropriate to request a coagulation screen for patients receiving warfarin or heparin as the reference ranges will not be valid. If monitoring of anticoagulants required select the appropriate test (warfarin monitoring or heparin monitoring).

Anticoagulant monitoring:

Always ensure the request form contains details of any anticoagulants given.

All INR results >4.0 will be telephoned to the patient by specialised anticoagulant nurses. No other INR results are routinely telephoned.

For queries related to the anticoagulant monitoring service contact the anticoagulation service.

VTE (venous thromboembolism) screening:

D-dimer is used to help venous thromboembolism. It is a restricted test and should only be used in conjunction with a clinical probability score e.g. Wells score and when the result will affect patient management (e.g. allow VTE to be excluded without radiology tests). D-dimer results are generally raised in in-patients and should not be requested.

Specialist Coagulation Tests:

Discuss with a Consultant Haematologist.

|TESTS |SPECIMEN |TURNAROUND |SPECIAL |KEY FACTORS |

| |REQUIRED |TIMES |INSTRUCTIONS |AFFECTING TEST PERFORMANCE OR |

| | | | |INTERPRETATION |

|Clotting Screen (PT + APTT) | | | |All coagulation tests require a good|

| |3.5ml blue top |3 hours | |clean venepuncture to avoid sample |

| |citrate | | |activation |

|INR (oral anticoagulation) |3.5ml blue |3 hours | | |

| |top citrate | | | |

|APTT ratio (unfractionated |3.5ml blue |3 hours | | |

|heparin) |top citrate | | | |

|Anti-factor Xa (Low |3.5ml blue |3 hours | | |

|molecular weight heparin) |top citrate | | | |

| |3.5ml blue |3 hours | | |

|D-Dimer (DD) |top citrate | | | |

| |3.5ml blue |3 hours | | |

|Fibrinogen (FIB) |top citrate | | | |

| |3.5ml blue |3 hours | | |

|Thrombin time (TT) |top citrate citrate | | | |

| |1.4 ml |3 hours | | |

| |citrate paediatric | | | |

|Clotting tests (paediatric) |blue tube | | | |

|Coagulation factors |2 x 3.5ml |Weekly |Discuss with | |

|(VIII,IX,XI,XII,) |blue top citrate | |Haematologist | |

| |4 x 3.5ml |2 weeks | |Indicate if patient on |

|Lupus anticoagulant screen |blue top citrate | | |anticoagulants |

| |4 x 3.5ml |2 weeks |See guidelines |Patient should be |

|Thrombophilia screen (AT3, PC, FPS, |blue n top citrate | |below |minimum 1 month post anticoagulant |

|APC,) | | | |therapy or post thrombotic episode |

Coagulation reference ranges

Adult normal range

|Test |Ref/Normal Range |Alert/Critical Value |Source of reference range |

| |(+/- 2SD) | | |

|INR |0.8-1.2 |6 |ED-HAE-EssentialBloodCoag |

|APTT |0.8-1.2 |4 |ED-HAE-SynthASil |

|(APTR) | | |ED-HAE-Synthasil |

| | | |LI-HAE-APTRConfirm |

| | | |ED-HAE-PracHemThrom2ndED |

|Fibrinogen-Clauss |1.5 – 4.0g/L |Reflex to low or high curve. |ED-HAE-BSHGuideFib |

|TT |10-17 seconds |17 |HemosIL Thrombin Time reagent kit insert |

| | | |ED-HAE-ACLThrombin |

| | | |ED-HAE-ThrombinTime |

|D-Dimer |2.2 | |LR-HAE-ACL-APCRassayCutOff |

|Protein C |70-140% | |ED-HAE-HemosILProtC |

|Free Protein S |Male = 74-146 % | |ED-HAE-ACLFreeProteinS |

| |Female = 55-124 % | | |

|Lupus DRVVT screen |A polymorphism |

|Synonyms |PT20210 F2 Leiden (FII Leiden) |

|Sample type |1 x EDTA Mauve top) |

|Test instructions |Transport at ambient temperature. |

|Referral laboratory |Molecular Haemostasis Laboratory |

| |St Thomas' Hospital |

| |North Wing - 4th and 5th Floors |

| |Westminster Bridge Road |

| |London SE1 7EH |

| |02071882798 |

|Turnaround time |7 days |

|Test indications |Increased thrombophilia risk. |

| |Increased plasma Prothrombin levels. |

|Reference intervals |Contact referral laboratory |

|Interferences | |

|Assay |Taipan snake venom time |

|Synonyms |TSVT |

|Sample type |2 X Citrate (blue top) |

|Test instructions |The samples should be analysed or manipulated & stored in the |

| |laboratory within 4 hours of venepuncture. |

|Referral laboratory |Diagnostic Haemostasis and Thrombosis Department |

| |020 7188 2797 |

| |St Thomas' Hospital |

| |North Wing - 4th and 5th Floors |

| |Westminster Bridge Road |

| |London SE1 7EH |

|Turnaround time |2 Weeks |

|Test indications |Antiphospholipid syndrome, acquired thrombophilia |

|Reference intervals |TSVT: 0.87 – 1.14 |

| |ET: 0.88 – 1.14 |

|Interferences | |

|Assay |von Willebrand factor |

|Synonyms |vWF |

|Sample type |Citrate (Blue top) |

|Test instructions |The sample should be analysed or manipulated & stored in the |

| |laboratory within 4 hours of venepuncture |

|Referral laboratory |Diagnostic Haemostasis and Thrombosis Department |

| |020 7188 2797 |

| |St Thomas' Hospital |

| |North Wing - 4th and 5th Floors |

| |Westminster Bridge Road |

| |London SE1 7EH |

|Turnaround time |Contact laboratory |

|Test indications |Family history of von Willebrand Disease, investigation of bleeding |

| |tendency |

|Reference intervals |N/A |

|Interferences | |

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