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Laboratory Diagnosis. What is laboratory diagnosis? Laboratory diagnosis is such a diagnostic process in which the samples coming from patients’ blood,

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Presentation on theme: "Laboratory Diagnosis. What is laboratory diagnosis? Laboratory diagnosis is such a diagnostic process in which the samples coming from patients’ blood,"— Presentation transcript:

1 Laboratory Diagnosis

2 What is laboratory diagnosis? Laboratory diagnosis is such a diagnostic process in which the samples coming from patients’ blood, body fluid, secretion, excretion, tissues and cells are examined by using various laboratory methods to get useful data which may reflects body` s functional status, pathological changes and etiological hints.

3 Why to study laboratory diagnosis? l Laboratory diagnosis is a very important part in diagnostics. After we have finished the study of physical diagnosis and instrument examination, we might be able to judge what kind of disease this patient probably suffer from. For further diagnosis, it is still not enough if we only master physical examination. We have to depend upon some laboratory data to ensure our diagnosis.

4 How to learn laboratory diagnosis? l First, the methods used in laboratory diagnosis are more automatic and computerized. l Second, the aim is to train for clinical application. l For each test, referential values and clinical significance always should be known very well.

5 Chapter 1 Blood Test Blood routine test (BRT) l 1. content (items) of BRT l red blood cell count (RBC) l hemoglobin(HB)concentration determination l total white blood cell count (WBC) l leukocyte differential count (DC) l platelet count (PC) l 1. content (items) of BRT l red blood cell count (RBC) l hemoglobin(HB)concentration determination l total white blood cell count (WBC) l leukocyte differential count (DC) l platelet count (PC)

6 R ferential values of BRT l Table 1. Deferential values for blood routine test in different age and sex l ___________________________________ l male female newborn l ___________________________________ RBC 4.0 - 5.5 3.5 - 5.0 6.0 - 7.0 HB 120 - 160 110 - 150 170 - 200 WBC 4 - 10 4 - 10 15 - 20 PLT 100 - 300 100 - 300 l ___________________________________

7 Table 2 Referential values of differential count l ___________________________________ l type percent value(%) absolute value ___________________________________ N st 1 - 5 0.04 - 0.5 N se 50 - 70 2 - 7 LYM 20 - 40 0.8-4 Mo 3 - 8 0.12 - 0.8 Eo 0.5 - 5 0.02 - 0.5 l Ba 0-1 0-0.1 l ___________________________________

8 Morphology of blood cells l Red blood cell: normocyte: 6-9um, discocyte- shape, no nucleus l white blood cell divided in five types: neutrophilic granulocyte : neutrophil stab cell (Nst), neutrophil segmented cell (Nse), lymphocyte, basophil granulocyte, eosinophil granulocyte l Platelet: smallest cell in peripheral blood

9 In automatic blood cell analyzer, 25 items are included together : l WBC : 4.0~10.0x109/L l LYM : 0.8~ 4.0 x109 /L (20~40%) l MID( middle cells) : monocyte, eosinocyte, basocyte, immature cells(rare be seen) l GRAN(granulocyte):2.0~7.8x109/L(50~70%) RBC:4.0~5.5x1012/L(male),3.5~5.0x 1012 /L (female) l HGB: 120~160 g/L (male), 110~150 g /L (femal)

10 - l HCT(hematocrit):0.4~0.5L/L(male), 0.37~ 0.48 L/L (female) l MCV ( mean corpuscular volume): 82~95 fl l MCH(mean corpuscular hemoglubin): 27~31 pg l MCHC ( mean corpuscular hemoglubin concentration): 320~360 g/L l RDW ( red cell distribution width ): 11.5~14.5% PLT( platelet): 100~300 x 109 /L l MPV ( mean platelet volume): 8.7~16.5 fl l PDW ( platelet distribution width): 15.8~21.4%

11 Clinical significance l Anomalies of RBC and HB l 1. decrease of RBC and HB l Anemia: When RBC and HB of individual is lower than the referential values of the people in same age, same sex and same area. Usually, if Hb of adult male and female is lower than 120g/L or 110g/L respectively, they are considered as anemia.

12 According to the level of Hb, anemia is divided into four degrees in clinic l mild anemia: Hb < 120g/L(male) or < 110g/L(female) l medium anemia : Hb <90 g/L l severe anemia : Hb <60g/L l extreme severe anemia: Hb <30g/L

13 Physiological anemia l infant and children aged between 3 months to 15 years, l pregnant women in middle or terminal periods of pregnancy, l elderly people.

14 Pathological anemia. l decrease in synthesis of red blood cell in bone marrow l increased destroy of red blood cell in peripheral blood l blood loss

15 Decrease in synthesis of red blood cell in bone marrow l disorder of hemopoiesis in bone marrow: aplastic anemia l infiltration of bone marrow by tumorous cell: leukemia, multiple myeloma, lymphoma, metastatic tumor l deficiency of hemopoietic materials or factors: iron deficiency anemia(IDA), megaloblastic anemia (folic acid deficiency)

16 Destroy of RBC in peripheral blood -- hemolytic anemia l Hereditory disease : hereditory sphrocytosis (HS), G6PD deficiency, thalassmia, Hb disease l Acquired anomalies : hemolysis by immunological, physical, chemical, biological and mechanical factors

17 Blood loss -- hemorrhagic anemia l acute blood loss : acute upper digestive tract bleeding, splenic rupture l chronic blood loss : hemorrhoid, hookworm disease, hypermenorrhea, GI tumor

18 2.Increase of RBC and Hb Comparative increase of RBC and Hb: due to the decrease of volume of plasma-- severe dehydration: severe vomoting, diarrhea, severe burn Absolute increase of RBC and Hb—ery- throcytosis: polycythemia vera(PV), chronic cardiopulmonary diseases: cor pulmonale, obstructive emphysema, scarcity of oxygen.

19 Anomalies of WBC and DC In most cases, increase and decrease of WBC is chiefly depend on the numbers of neutrophil granuiocyte in the blood. Leukocytosis: WBC is high than 10 x10 9 /L Leukopenia: WBC is lower than 4 x10 9 /L

20 1. Neutrophil granulocyte A. granulocytosis: 5 causes leading to granulocytosis acute infection or inflammation: acute pyogenic tonsilitis, acute appendicitis, hematosepsis damage or necrosis of tissue: severe burn, acute myocardiac infarction

21 acute blood loss: digestive tract bleeding, splenic rupture Acute poisoning: uremia, ketoacidosis Malignant blood diseases and tumor: leukemia, metastatic tumor

22 B.Granulocytopenia: leukopenia: WBC < 4 x10 9 /L granulocytopenia: neutrophil granulocyte < 1.5 x10 9 /L agranulocytopenia: neutrophil granulocyte < 0.5 x10 9 /L

23 5 causes for granulocytopenia: Infection of bacteria and virus: typhoid, influenza, measles some physical and chemical factors: drug autoimmune diseases: systemic lupus erythematosis (SLE), malignant blood diseases: aplastic anemia,leukemia hypersplenism

24 Changes of nucleus nucleus shift to left: mild shift to left: Nst > 6% only medium shift to left: Nst > 10% with meta- myelocyte severe shift to left : Nst > 25% with more immuture cell (leukemiod reaction ) nucleus shift to left cab be seen in acute infection, acute poisoning, acute hemolysis

25 Nucleus shift to right: multi-segmented Nse >3% 3-lobed Nse is normal segmented neutrophil in BP nucleus shift to right can be seen megaloblastic anemia administration of anti-metabolic drugs

26 2. lymphocyte A. Lymphocytosis: DC: Lym > 40% Virus infection: infectious mononucleosis, chickenpox, Lymphocytic leukemia: ALL, AA, lymphoma GVHD or GVHR after BMT

27 B. lymphocytopenia: administration of anti-tumor drugs, prednision radiation: x-ray, r-ray, isotope 3. monocytosis: malaria, black fever, TB, subacute bacterial endocarditis(SBE) monocytic leukemia, malignant histocytosis,

28 4. eosinophilia allergic diseases, hypersensitive diseases parasites infection: Skin diseases: psoriasis blood diseases: CML, eosinophilic leukemia, lymphoma Infectious diseas: scarlatina Hypereosinophilic syndrome

29 5. Basophil granulocyte bosophilia: CML, basophilic leukemia, myelofibrosis

30 Other tests for red blood cell 1. Reticulocyte count (RC) 0.05-0.015 (0.5-1.5%) 24-84 x10 9 /L l Reticulocytosis: hemolytic anemia acute hemorrhagic anemia index of therapeutic effect: IDA, megaloblastic anemia

31 Reticulocytopenia: Aplastic anemia leukemia

32 2. Hematocrit(Hct) 0.4-0.5L/L(male), 0.37-0.48L/L(female) increase of Hct: blood concentration water loss polycythemia(PV) decreas of Hct: anemia

33 3. Mean values of RBC Mean corpuscular volume (MCV): MCV=Hct/RBC 82-95fl Mean corpuscular hemoglubin(MCH): MCH=Hb/RBC 27-31pg Mean corpuscular hemoglubin concentration MCHC=Hb/Hct 320-360g/L

34 Morphological classification of anemia Classification MCV MCH MCHC diseases Normocytic 82-95 27-31 320-360 AA, HA, leukemia Macrocytic >100 > 31 320-360 MA, pernicious anemia Microcytic < 80 < 27 320-260 infection, tumor, uremia Microcytic < 80 < 27 < 320 IDA, thalassemia Hypochromic sideroblastic anemia

35 4.Erythrocyte sedimentation rate(ESR) 0-15mm/h (male), 0-20mm/h (female) higher ESR: infection and inflammation: rheumatic diseases,tuberculosis malignant tumor anemia damage or necrosis of tissue globulinemia, cholesterolemia

36 Chapter 2. Bone Marrow Examination 1. Clinical application of marrow examination l Diagnosis for hematopoietic system diseases: leukemia, myeloma, aplastic anemia, etc l parasite infectious diseases: malaria, black fever l metabolic diseases: Gaucher disease. Niemann-Pick disease

37 Indications: Fever origin unknown (FOU) Cachexia Hepatomegaly, splenomegaly, lymphoadenovarix Abnormal in quantity and quality in peripheral blood cell

38 Contraindication: Hemophilia Pyogenic infection in local skin Methods: bone marrow aspiration bone marrow biopsy

39 2.Development of blood cells myeloblast promyelocyte myelocyte metamyelocyte Nst Seg normoblast basophilic normoblast polychromatic normoblast orthochromatic normoblast erythrocyte

40 Monoblast promonocyte monocyte macrophage histiocyte Megakaryoblast promega granular Meg thrombocytogenous Meg platelet B-lymphoblast B-prolymphoblast B- lymphocyte plasmablast proplasmacyte plasmacyte

41 3. Regulation of blood cell development l Cytobody: from large to small, Mega is an exception l Cytoplasma: from less to more, color from blue to pink or orange red, granules from invisible to visible, granules from non-specific to specific

42 l Nucleus: from large to small, Mega is an exception, shape of nucleus from round to irregular, chromatin patten of nucleus from fine to coarse. nucleolus from existance to nonexistance l Ration of nucleus to cytoplasma: from bigger to smaller



45 5.Contene and step of marrow examination l Myelogram: l Low power(LP): Wright stain of marrow film plastic degree: 5 classification Meg count : 7-35/1.5x3cm2 ousspecial cell: metastatic tumorous cell

46 Oil immersion: Nucleated cell count (%): 200-500 Ratio of myeloid to erythroid (M:E): 2-4:1 Morpholigical description of every series Special cells and parasite Diagnosis

47 Hemogram: Low power(LP): Oil immersion len: differential count 100 leukocyte (DC) Immature cell (nucleated cell) parasites

48 6. Normal myelogram and hemogram 1. Myelogram l Normal cellularity: erythrocyte: nucleated cell l M:E=2-4:1 l Granulocytic series is 40-60% in all nucleated cells l Erythrocytic series is 20% in all nucleated cells l Lymphocyte is 20%, monocyte is <4% in all nucleated cells l Meg count is 7-35, platelet is normal l Other cell: plasmacyte and histiocyte are rare l No specific cells and parasite

49 Plastic degree of marrow Plastic degree erythrocyte: nucleated diseases Extreme hypercellularity 1:1 leukemia Significant hyper- 10:1 leukemia, ITP, Cellularity plastic anemia Normal cellularity 20:1 anemia, normal myelogram Hypocellularity 50:1 AA,agranulocy- topenia Extreme hypocellularity 300:1 aplastic anemia

50 2. Hemogram: DC is normal No nucleated red blood cell No immature white cell Platelet is normal



53 7. Characters of common blood diseases in microscope l Iron deficiency anemia(IDA) l Aplastic anemia(AA) l Acute leukemia(AL) l Chronic myelocytic leukemia(CML) l Idiopathic thrombocytopenia purpura(ITP)

54 1. Iron Deficiency Anemia(IDA) l Hemogram: Hb, RBC, normal RC, RBC hypochromia, exaggeration of central pallor to from rings, anisocytosis normal WBC and platelet

55 Myelogram: Significant hypercellularity M:E Erythroid hyperplasia with small normoblast Pycnotic nuclei and scanty cytoplasma irregular margin


57 2. Aplastic Anemia(AA) l Hemogram: pancytopenia: Hb,RBC, WBC,PLT RC <0.005 or absolute value < 15 x10 9 /L RBC is normocytic

58 Myelogram: Hypocellularity or extreme hypocellularity, Hematopoietic cell decrease: erythroid, myeloid cells, meg or absent, platelet rarely be seen Non-hematoietic cell increase: lymphocyte Reticular cell, plasmacyte


60 3. Acute Leukemia(AL) l FAB classification: acute lymphocytic leukemia: L1-L3 acute non-lymphocytic leukemia: M0-M7 l Hemogram: Hb, RBC WBC uncertain: normal, increase, decreas platelet:

61 Myelogram: Extreme or significant hypercellularity M:E Hyperplasia of certain line : Myeliod(M0,M1-3), monocyte(M4-5), meg(M7), lymphocyte(L1-3), erythroid(M6) Depressed erythroid and meg line













74 4. Chronic Myelocytic Leukemia(CML) l Hemogram: WBC increase rxtremly DC: E0, Ba increas, myelocyte, metamyelocyte, band cell increase normal Hb, RBC or mild decrease normal plt

75 Myelogram: Extreme hypercellularity M:E increase Extreme hyperplasia of granulocyte with increas of myelocyte, metamyelocyte, band cell, E0, Ba Normal erythroid and Megakaryocyte




79 5. Idiopathic Thrombocytopenic Purpura(ITP) l Hemogram: normal Hb, RBC normal WBC and DC PC decreas with abnormal morphology

80 Myelogram: Significant hypercellulerity or normal cellularity Normal myeloid, erythroid line Significant hyperplasia of Meg. Impaired maturation of Meg: Granular Meg or promeg increase Platelet producting Meg absent Platelet rarely be seen


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