Laboratory for diagnosis of THALASSEMIA

Transcription

1 SCBM343 CLINICAL PATHOLOGY 2(1-2-3) Laboratory for diagnosis of THALASSEMIA PORNTHIP CHAICHOMPOO

2 Acknowledgements Dr. Pranee Winichagoon Fucharoen Ms. Pornnapa Khampan Thalassemia Research Center Institute of Molecular Biosciences Mahidol University

3 Objectives Interpretation of blood smear from thalassemic blood samples for grade red blood cell morphology and color staining, grade amounts of platelets, differential white blood cells (neutrophils, lymphocytes, monocytes, eosinophils and basophils) and nucleated red blood cells. Diagnosis of thalassemic carriers and thalassemic diseases from complete blood count, blood smear and hemoglobin typing

4 Blood smear analysis for Diagnosis of Anemia & Thalassemia

5 Interpretation of red blood cells (RBCs) on film smear by grading Count 10 fields of Oil Immersion (100x) Examination Grading: Found (Mean) 0 5% 5 9% 10 24% 25 49% 50 74% % Grade Normal Slight Color Normochromia / Hypochromia / Hyperchromia Size Normocyte / Microcyte / Macrocyte Shape Biconcave / Target cell / Tear drop / Echinocyte / Ovalocyte / Fragmented cells / Other if found (specific) such as polychromasia Comments Anisocytosis / Poikilocytosis / Anisopoikilocytosis

6 Interpretation of platelets on film smear by grading Count 10 fields of Oil Immersion (100x) Examination Grading: Found (Mean) <10 cells / field cells / field > 30 cells / field Grade Decrease Adequate Increase Estimated platelet count / µl = Average count in 10 fields x 15,000 Comments Giant platelet / Clumps

7 Interpretation of white blood cells (WBCs) and nucleated RBCs (NRBCs) on film smear Count 100 WBCs of Oil Immersion (100x) Examination Report as % of WBCs In during count 100 WBCs, how many NRBCs are found Calculation of corrected WBCs (cells / µl) Corrected WBCs (cells / µl) = WBC count x number of NRBCs in during count 100 WBCs Comments Corrected WBCs Neutropenia / Neutrocytosis Lymphopenia / Lymphocytosis

8 Blood smear report STUDENT NAME STUDENT ID SPECIMEN CODE RBC morphology: Color Normochromia Hypochromia Hyperchromia Size Normocyte Microcyte Shape, give score of grading Biconcave.. Target cell.. Tear drop... Echinocyte... Ovalocyte... Fragmented cells... Macrocyte Other if found (specific) Comments:. Platelets: Number per field:. Estimated platelet count (cells/µl):. Decrease Adequate Increase Comments:..

9 Blood smear report (cont.) Differential WBCs: % Neutrophils % Lymphocytes % Monocytes % Eosinophils % Basophils NRBC count: NRBCs/ 100 WBCs If WBC count from CBC is 15,000 cells/µl, how many corrected WBC count.. (show your calculation below) Calculation: Comments:..

10 Worksheet for differential WBC and NRBCs count Column # Total Neutrophil Lymphocyte Monocyte Eosinophil Basophil NRBCs How to use this table: Each column must contain 10 cells. These 10 cells are neutrophil or lymphocyte or monocyte or eosinophil or basophil. Total 10 column x 10 cells per column = 100 cells as called 100WBCs. In during, 100 WBCs are counting. If NRBCs are found, note into the yellowed cells.

11 Diagnosis of thalassemic carriers and thalassemic diseases from - complete blood count and - hemoglobin typing

12 Thalassemias Thalassemic Diseases 1. Homozygous b-thalassemia 2. b-thalassemia/hb E 3. Hb Bart, s hydrops fetalis 4. Hb H disease

13 P1 x a-thalassemia 1 a-thalassemia 2 F1 Hb H diesease a-thalassemia 1 a-thalassemia 2 Normal

14 P1 x a-thalassemia 1 a-thalassemia 1 F1 Hb Bart s hydrops fetalis a-thalassemia 1 a-thalassemia 1 Normal

15 P1 x b E b-thalassemia trait HbE trait F1 b E b E b-thalassemia/hbe disease b-thal. trait HbE trait Normal

16 Thalassemia Diagnosis Hematological Data J Hb J MCV J MCH J MCHC J RDW Hb Typing Data Hb Type Hb A 2 (E) Bart s F A A 2 /E CS/PS Hb F Quantitative Hb A Hb Bart s Hb CS/PS Qualitative

17 Interpretation for diagnosis of thalassemia MCV (fl) MCH (pg) Hb typing %HbA2(E) %HbF Normal A2A a-thalassemia 2 trait A2A a-thalassemia 1 trait A2A b-thalassemia trait A2A Hb E trait EA Homozygous HbE EE b-thalassemia/hbe dis EF HbH disease A2ABart sh

18 Red Cell Indices Normal a-thal 2 Normocyte Hb Analysis Microcyte Hb Analysis Thalassemic disease (specified by Hb types) A 2 + A A+CS A+E EE A 2 + A Hb Constrant Hb E Homozygote Spring Hb E Determination Hb A 2 Determination E ~ 25-30% E 21% or less A 2 < 3.5% A 2 > 3.5% Hb E trait Hb Bart s b-thal trait a-thal 2/Hb E Hb Bart s Hb < 10 g/dl Hb > 10 g/dl Hb < 10 g/dl Hb > 10 g/dl Hb E with a-thal 1/Hb E Fe deficiency or a-thal 1 trait Fe deficiency Fe deficiency Homo a-thal 2 on top of thalassemia trait

19 Case study (example) Family 1 Mr. Z x Mrs. Z Risk couple?

20 Mr. Z Hematological Data A Hb (g/dl) 12.4 MCV (fl) 90.5 MCH (pg) 29 MCHC (g/dl) 31.5 RDW (%) 12.1 HbF HbA HbA 2 A 2 Family 1: 1. Individual diagnosis (Male and Female) by give %of each Hb type, interpretation of Hb typing and diagnosis for thalassemia 2. Give possibility of this couple to give a new birth. Are they a risk couple for thalassemia disease?

21 Mr. Z A Hematological Data Hb (g/dl) 12.4 MCV (fl) 90.5 MCH (pg) 29 MCHC (g/dl) 31.5 RDW (%) 12.1 HbF HbA HbA 2 A 2 Hemoglobin Typing Hb type A 2 A Hb A 2 (E) 2.8 % Hb F 0.3 % Hb A 87.5 % Diagnosis: Normal Hb typing (aa/aa, b N /b N )

22 Mrs. Z F E Hematological Data Hb (g/dl) 9.4 MCV (fl) 68.0 MCH (pg) 21.5 MCHC (g/dl) 31.6 RDW (%) 23.0 HbF HbE A 2 Hemoglobin Typing Hb type Hb A 2 (E) Hb F Hb A

23 Mrs. Z F E Hematological Data Hb (g/dl) 9.4 MCV (fl) 68.0 MCH (pg) 21.5 MCHC (g/dl) 31.6 RDW (%) 23.0 HbF HbE A 2 Hemoglobin Typing Hb type EF Hb A 2 (E) 52.7 Hb F 43.2 Hb A - Diagnosis: b-thalassemia/hbe disease (aa/aa, b 0 /b E )

24 Family 1 Mr. Z Mrs. Z Normal Hb typing b-thal/hbe disease b E b-thal trait Hb E trait b-thal trait Hb E trait b E b E

25 Case study (example) Family 1 Mr. Z x Mrs. Z Risk couple?

26 Activities for Lab Individual word Choose 3 specimen blood smear with Code Nxxx, NSxxx and Sxxx for any number. Specify No.(xxx) on report. Choose one set of case study. There are 4 sets. Each set contain 2 family (4 subjects). Report individual diagnosis of thalassemia (4 subjects), family pedigree chart and answer the question that they are a risk couple or not. Submit report at the end of class Evaluation: 100% of lab examination on L4 Anemia & Thalassemia