Successfully reported this slideshow.
We use your LinkedIn profile and activity data to personalize ads and to show you more relevant ads. You can change your ad preferences anytime.
Laboratory Diagnosis of AnemiaBy
Dr. Varughese George
Department of Pathology
Learning Objectives
At the end of this briefing, you should know
• Clinical presentation of anemia.
• Definition of anemia...
Clinical presentation of anemias
• Fatigue and weakness
• Headache
• Tinnitus
• Numbness and coldness
• Pallor
• Dyspnea a...
Clinical presentation of anaemias
Initial Laboratory Work-Up
• Hemoglobin concentration.
• Packed Cell Volume.
• Red cell indices.
• Peripheral Blood Smear.
Definition of Anaemia
• Anaemia is defined as a reduction in the concentration
of circulating haemoglobin below the level ...
Blood parameters (Laboratory Normal Range)
Blood parameters (Laboratory Normal Range)
•In full term infants, hemoglobin is 18.0 ± 4.0 g/dl
•Children - 12.5 ± 1.5 g/d...
Grading of Anaemia
Classification of Anaemias
• Morphological Classification
• Etiological Classification
• Classification based on reticuloc...
Morphological Classification of Anemias
Hematocrit
• Proportion of the volume of red cells relative
to the volume of blood
• Rules of Three:
– RBC X 3 = Hemoglobi...
Mean Corpuscular Volume
• Dividing the total volume of red cells by the
number of red cells
• Index for average size of re...
Mean Corpuscular Hemoglobin
• Average amount of haemoglobin in each red
cell.
• It is expressed in picograms or pg.
• Norm...
Mean Corpuscular Hemoglobin Concentration
• This represents the average concentration of
haemoglobin in a given volume of ...
Red cell distribution width
• Variation in red cell size
• Normal range - 12.8% ± 1.2%
• Low in B-thalassemia trait
• High...
17
1 Microcytic/hypochromic
3
1 2
2 Macrocytic/Normochromic
3 Normocytic/Normochromic
Morphologic Categories of Anemia
N.B...
Macrocytic anemia
• Low/normal reticulocyte
count, macrocytosis(oval
and round)
• Elevated MCV,MCHC
• Basophilic stippling...
Normocytic Normochromic anaemia
with effective erythropoiesis
Etiological Classification of Anaemias
Basic Approach to a diagnosis of anemia
Evaluation of microcytic hypochromic anaemia
Evaluation of macrocytic anaemia
Evaluation of normocytic anaemia
Evaluation of haemolytic anaemia
A simplified approach to diagnosis of haemolytic
anaemias
Reticulocyte Count
(In the Diagnosis of Anemia)
• Reticulocytes are non-
nucleated RBCs that still
contain RNA.
• Visualiz...
Fe++ deficiency anemia
• Low hemoglobin and low packed cell volume
• Low MCV,MCH and MCHC
• Microcytosis & hypochromia are...
31
Megaloblastic Anemia
Mild to severe anemia,
– Increased MCV & MCH, normal MCHC
– Low RBC, HGB, WBC and PLT counts (frag...
32
Macrocytic Ovalocytes
Blood NRBC Blood
Howell-Jolly body
Teardrop
Schistocyte
Stippled RBC &
Cabot Ring
Giant Platelet
...
Tests
• Folate and B12 levels
• Schilling test may be useful to establish
etiology of B12 deficiency
– Assesses radioactiv...
Anemia of chronic disease
• Normocytic anemia with ineffective erythropoiesis
(reduced reticulocyte count)
• Normochromic
...
Anemia of chronic disease
• Decreased serum iron,decreased total iron
binding capacity and normal or raised ferritin
• Inc...
Normochromic, normocytic anemia with
effective erythropoiesis
INCREASED reticulocyte count
• Acute blood loss
– Very acute...
Aplastic anemia
• Pancytopenia caused by bone marrow
failure…decreased production of all cell
lines and replacement of mar...
Hemolytic anemia
• Abnormality intrinsic to red cells-
1. Hereditary spherocytosis
2. Thalassamia
3. Sickle cell anaemia
4...
Evaluation of haemolytic anaemia
Hereditary spherocytosis
• Inherited defect in the red cell membrane
cytoskeleton (spectrin, ankyrin or band 3)
leading to...
Thalassemia
• Decreased or absent globin
chains
• Alpha and beta
thalassemias
• Microcytic
hypochromic,target
cells,basoph...
Sickle cell anaemia
• Presence of Hb S
• Point mutation in 6th place of beta
chain
• Substitution of valine for glutamic a...
Sickle cell anaemia
• Sickling test is positive.
• Solubility test is positive.
• Electrophoresis shows HbS.
• In sickle c...
Glucose-6-phosphate dehydrogenase
deficiency
• X linked disorder
• Reduced activity of G6PD
• Inability to remove H2O2
• A...
Glucose-6-phosphate dehydrogenase
deficiency
• Asymptomatic
• Neonatal jaundice
• Acute hemolytic anaemia
• Chronic hemoly...
Glucose-6-phosphate dehydrogenase
deficiency
• Screening tests-fluorescent spot
test,methemoglobin reduction test and dye
...
Immune hemolytic anaemia
• Warm antibody-persons over 50 years,mild
jaundice and splenomegaly,red cells coated
with IgG,sp...
Coomb’s test
• Detects presence of either antibody on RBC or
of antibody in serum
• Helpful in determining if a hemolytic ...
SUMMARY
• Microcytic hypochromic anaemia-iron
deficiency
• Macrocytic hyperchromic-megaloblastic
anaemia
• Normochromic no...
Laboratory diagnosis of anemia
Laboratory diagnosis of anemia
Laboratory diagnosis of anemia
Upcoming SlideShare
Loading in …5
×

Laboratory diagnosis of anemia

19,583 views

Published on

Laboratory diagnosis of anemia

Published in: Health & Medicine
  • Have you ever used the help of ⇒ www.WritePaper.info ⇐? They can help you with any type of writing - from personal statement to research paper. Due to this service you'll save your time and get an essay without plagiarism.
       Reply 
    Are you sure you want to  Yes  No
    Your message goes here
  • Hi there! I just wanted to share a list of sites that helped me a lot during my studies: .................................................................................................................................... www.EssayWrite.best - Write an essay .................................................................................................................................... www.LitReview.xyz - Summary of books .................................................................................................................................... www.Coursework.best - Online coursework .................................................................................................................................... www.Dissertations.me - proquest dissertations .................................................................................................................................... www.ReMovie.club - Movies reviews .................................................................................................................................... www.WebSlides.vip - Best powerpoint presentations .................................................................................................................................... www.WritePaper.info - Write a research paper .................................................................................................................................... www.EddyHelp.com - Homework help online .................................................................................................................................... www.MyResumeHelp.net - Professional resume writing service .................................................................................................................................. www.HelpWriting.net - Help with writing any papers ......................................................................................................................................... Save so as not to lose
       Reply 
    Are you sure you want to  Yes  No
    Your message goes here
  • Writing a good essay isn't easy and it's the fruit of hard work. For help you can check essay writing expert. Check out, please HelpWriting.net I think they are the best
       Reply 
    Are you sure you want to  Yes  No
    Your message goes here
  • I would absolutely recommend this program. You get lots of support and tools, and you get to be open and share, but you never feel embarrassed or ashamed. Everyone is so accepting and kind. It's just a wonderful community. Joining the program was the best thing I did to help my recovery. ♥♥♥ http://t.cn/A6Pq6ilz
       Reply 
    Are you sure you want to  Yes  No
    Your message goes here
  • I purchased your e-book, How to Reverse Diabetes. This is really an amazing life-saving book, it's a blessing beyond words. I've downloaded your program because my dad has been suffering from diabetes for a very long time but now, thanks to your help, his blood sugar level is well within normal range and all his symptoms are gone. I cannot explain in words how much this book has meant to me and my family. I just want to let you know that your program has totally changed my life; you truly are a blessing for humanity. People need to hear your message because it can save many many people’s lives. The world needs more people like you. From the bottom of my heart, thank you thank you thank you! ♣♣♣ https://bit.ly/2swQ6OO
       Reply 
    Are you sure you want to  Yes  No
    Your message goes here

Laboratory diagnosis of anemia

  1. 1. Laboratory Diagnosis of AnemiaBy Dr. Varughese George Department of Pathology
  2. 2. Learning Objectives At the end of this briefing, you should know • Clinical presentation of anemia. • Definition of anemia. • Approach to diagnosis of anemia • Classification of anaemia • Distinct features of each type of anemia.
  3. 3. Clinical presentation of anemias • Fatigue and weakness • Headache • Tinnitus • Numbness and coldness • Pallor • Dyspnea and palpitations • Angina pectoris • Intermittent claudication • Hemorrhages in the fundus of eyes • Menorrhagia • Anorexia • Flatulence • Nausea • Constipation
  4. 4. Clinical presentation of anaemias
  5. 5. Initial Laboratory Work-Up • Hemoglobin concentration. • Packed Cell Volume. • Red cell indices. • Peripheral Blood Smear.
  6. 6. Definition of Anaemia • Anaemia is defined as a reduction in the concentration of circulating haemoglobin below the level that is expected for healthy personsof same age and sex in the same environment. 6
  7. 7. Blood parameters (Laboratory Normal Range)
  8. 8. Blood parameters (Laboratory Normal Range) •In full term infants, hemoglobin is 18.0 ± 4.0 g/dl •Children - 12.5 ± 1.5 g/dl •Children(6-12 years) - 13.5 ± 2.0 g/dl
  9. 9. Grading of Anaemia
  10. 10. Classification of Anaemias • Morphological Classification • Etiological Classification • Classification based on reticulocyte response.
  11. 11. Morphological Classification of Anemias
  12. 12. Hematocrit • Proportion of the volume of red cells relative to the volume of blood • Rules of Three: – RBC X 3 = Hemoglobin – Hemoglobin X 3 = Hematocrit Packed cell volume (PCV) or Haematocrit (Hct) Men - 0.45 ± 0.05 l/l (40-50%) Women - 0.41 ± 0.05 l/l (38-45 % in non- pregnant women 36-42 % in pregnant women)
  13. 13. Mean Corpuscular Volume • Dividing the total volume of red cells by the number of red cells • Index for average size of red cells • Normal range - 92 ± 9 fl 13
  14. 14. Mean Corpuscular Hemoglobin • Average amount of haemoglobin in each red cell. • It is expressed in picograms or pg. • Normal range - 29.5 ± 2.5 pg
  15. 15. Mean Corpuscular Hemoglobin Concentration • This represents the average concentration of haemoglobin in a given volume of packed red cells. • Normal range – 330 ± 15 g/l • MCHC raised in hereditary spherocytosis. • Decreased in hypochromic anaemia.
  16. 16. Red cell distribution width • Variation in red cell size • Normal range - 12.8% ± 1.2% • Low in B-thalassemia trait • High in iron deficiency anaemia • Normal in anaemia of chronic disease
  17. 17. 17 1 Microcytic/hypochromic 3 1 2 2 Macrocytic/Normochromic 3 Normocytic/Normochromic Morphologic Categories of Anemia N.B. The nucleus of a small lymphocyte (shown by the arrow) is used as a reference to a normal red cell size
  18. 18. Macrocytic anemia • Low/normal reticulocyte count, macrocytosis(oval and round) • Elevated MCV,MCHC • Basophilic stippling • Howell-jolly bodies • Cabot rings • Pancytopenia • Hypersegmented neutrophils • Bone marrow- megaloblastic maturation,sieve like chromatin,nuclear- cytoplasmic asynchrony,maturation arrest
  19. 19. Normocytic Normochromic anaemia with effective erythropoiesis
  20. 20. Etiological Classification of Anaemias
  21. 21. Basic Approach to a diagnosis of anemia
  22. 22. Evaluation of microcytic hypochromic anaemia
  23. 23. Evaluation of macrocytic anaemia
  24. 24. Evaluation of normocytic anaemia
  25. 25. Evaluation of haemolytic anaemia
  26. 26. A simplified approach to diagnosis of haemolytic anaemias
  27. 27. Reticulocyte Count (In the Diagnosis of Anemia) • Reticulocytes are non- nucleated RBCs that still contain RNA. • Visualized by staining with supravital dyes, including new methylene blue or brilliant cresyl blue. • Useful in determining response and potential of bone marrow. • Normal range is 0.5-2.5% of all erythrocytes. 29
  28. 28. Fe++ deficiency anemia • Low hemoglobin and low packed cell volume • Low MCV,MCH and MCHC • Microcytosis & hypochromia are hallmarks • RDW is increased • Serum ferritin is less than 15 micro gram/dl • Serum iron is low,TIBC is increased and transferrin saturation is less than 10 percent • Free erythrocyte protoporphyrin is increased. • Increased soluble transferrin receptor in serum • Bone marrow-micronormoblastic,absence of stainable iron in bone marrow on Perls Prussian blue reaction
  29. 29. 31 Megaloblastic Anemia Mild to severe anemia, – Increased MCV & MCH, normal MCHC – Low RBC, HGB, WBC and PLT counts (fragile cells) due to ineffective hematopoiesis. – Low reticulocyte count – Macrocytic ovalocytes and teardrops; – Marked anisocytosis and poikilocytosis – Schistocytes/microcytes - due to RBC breakage upon leaving the BM – Erythroid hyperplasia - low M:E ratio (1:1) – Iron stores increased.
  30. 30. 32 Macrocytic Ovalocytes Blood NRBC Blood Howell-Jolly body Teardrop Schistocyte Stippled RBC & Cabot Ring Giant Platelet Pap bodies Hypersegmented Neutrophil >5 lobes Megaloblastic anemia
  31. 31. Tests • Folate and B12 levels • Schilling test may be useful to establish etiology of B12 deficiency – Assesses radioactive B12 absorption with and without exogenous IF • Other tests if pernicious anemia is suspected – Anti- parietal cell antibodies, anti-IF antibodies – Secondary causes of poor absorption should be sought (gastritis, ileal problems, etc.)
  32. 32. Anemia of chronic disease • Normocytic anemia with ineffective erythropoiesis (reduced reticulocyte count) • Normochromic • Results from – Chronic inflammation (e.g. rheumatologic disease): Cytokines released by inflammatory cells cause macrophages to accumulate iron and not transfer it to plasma or developing red cells (iron block anemia) – Inflammation – malignancy • Bone marrow suppression (EPO is elevated)
  33. 33. Anemia of chronic disease • Decreased serum iron,decreased total iron binding capacity and normal or raised ferritin • Increased marrow storage iron • ESR is high
  34. 34. Normochromic, normocytic anemia with effective erythropoiesis INCREASED reticulocyte count • Acute blood loss – Very acutely, with hypovolemia, may have normal blood counts, will become anemic with volume replenishment • Hemolytic anemia – Increased reticulocyte production cannot keep pace with loss of RBCs peripherally. • Response to specific therapy in nutritional anemias
  35. 35. Aplastic anemia • Pancytopenia caused by bone marrow failure…decreased production of all cell lines and replacement of marrow with fat. • Inherited- Fanconis anaemia, Dyskeratosis congenita • Acquired - Idiopathic,drugs like NSAIDs,chloramphenicol,benzene,parvo virus,hepatitis and EB virus.
  36. 36. Hemolytic anemia • Abnormality intrinsic to red cells- 1. Hereditary spherocytosis 2. Thalassamia 3. Sickle cell anaemia 4. Glucose -6-phosphate dehydrogenase deficiency • Abnormality extrinsic to red cells- 1. Immune 2. Mechanical etc
  37. 37. Evaluation of haemolytic anaemia
  38. 38. Hereditary spherocytosis • Inherited defect in the red cell membrane cytoskeleton (spectrin, ankyrin or band 3) leading to the formation of spherocytic red cells. • Autosomal dominant • Mild to moderate anaemia • Intermittent jaundice • Splenomegaly • Pigment gall stones • Peripheral smear-microspherocytes • Screening test-osmotic fragility
  39. 39. Thalassemia • Decreased or absent globin chains • Alpha and beta thalassemias • Microcytic hypochromic,target cells,basophilic stippling • Reticulocytosis • Hb F elevated in electrophoresis
  40. 40. Sickle cell anaemia • Presence of Hb S • Point mutation in 6th place of beta chain • Substitution of valine for glutamic acid • On deoxygenation,sickle cells are formed • Chronic hemolytic anaemia,vaso- occlusive crisis • Aplastic crisis • Hemolytic crisis • Infections
  41. 41. Sickle cell anaemia • Sickling test is positive. • Solubility test is positive. • Electrophoresis shows HbS. • In sickle cell trait, electrophoresis shows 60 percent of Hb A and 40 percent Hb S
  42. 42. Glucose-6-phosphate dehydrogenase deficiency • X linked disorder • Reduced activity of G6PD • Inability to remove H2O2 • Accumulated H2O2 leads to oxidation of hemoglobin with precipitation of globin chains • Heinz bodies • Red cells with heinz bodies destroyed in spleen(extravascular hemolysis)
  43. 43. Glucose-6-phosphate dehydrogenase deficiency • Asymptomatic • Neonatal jaundice • Acute hemolytic anaemia • Chronic hemolytic anaemia • On peripheral smear- polychromasia,fragmented red cells,spherocytes,bite cells,half ghost cells • Biochemical-increased bilirubin,hemoglobinemia and hemoglobinuria
  44. 44. Glucose-6-phosphate dehydrogenase deficiency • Screening tests-fluorescent spot test,methemoglobin reduction test and dye decolorisation test
  45. 45. Immune hemolytic anaemia • Warm antibody-persons over 50 years,mild jaundice and splenomegaly,red cells coated with IgG,spherocytes.Seen in autoimmune disorders,lymphoma • Cold antibody-acrocyanosis,IgM.Seen in cold agglutinin disease, Paroxysmal cold hemoglobinuria (PCH)
  46. 46. Coomb’s test • Detects presence of either antibody on RBC or of antibody in serum • Helpful in determining if a hemolytic anemia is immune-mediated
  47. 47. SUMMARY • Microcytic hypochromic anaemia-iron deficiency • Macrocytic hyperchromic-megaloblastic anaemia • Normochromic normocytic-hemolytic anaemia • Pancytopenia-megaloblastic and aplastic anemias

×