laboratory for diagnosis of thalassemia
TRANSCRIPT
SCBM343CLINICALPATHOLOGY2(1-2-3)
LaboratoryfordiagnosisofTHALASSEMIA
PORNTHIPCHAICHOMPOO
Acknowledgements
Dr.PraneeWinichagoon FucharoenMs.Pornnapa Khampan
ThalassemiaResearchCenterInstituteofMolecularBiosciencesMahidol University
Objectives
• Interpretationofbloodsmearfromthalassemicbloodsamplesforgraderedbloodcellmorphologyandcolorstaining,gradeamountsofplatelets,differentialwhitebloodcells(neutrophils,lymphocytes,monocytes,eosinophilsandbasophils)andnucleatedredbloodcells.• Diagnosisofthalassemiccarriersandthalassemicdiseasesfromcompletebloodcount,bloodsmearandhemoglobintyping
BloodsmearanalysisforDiagnosisofAnemia&Thalassemia
Interpretationofredbloodcells(RBCs)onfilmsmearbygrading
• Color• Normochromia /Hypochromia /Hyperchromia
• Size• Normocyte /Microcyte /Macrocyte
• Shape• Biconcave/Targetcell/Teardrop/Echinocyte/Ovalocyte /Fragmentedcells/
Otheriffound(specific)suchaspolychromasia• Comments
• Anisocytosis/Poikilocytosis /Anisopoikilocytosis
Grading: Found(Mean)
NormalSlight1+2+3+4+
Grade0– 5%5– 9%10– 24%25– 49%50– 74%75– 100%
Count10fieldsofOilImmersion(100x)Examination
Interpretationofplateletsonfilmsmearbygrading
• Comments• Giantplatelet/Clumps
Grading: Found(Mean)
DecreaseAdequateIncrease
Grade
<10cells/field10– 30cells/field>30cells/field
Count10fieldsofOilImmersion(100x)Examination
Estimatedplateletcount/µL=Averagecountin10fieldsx15,000
Interpretationofwhitebloodcells(WBCs)andnucleatedRBCs(NRBCs)onfilmsmear
• Comments• CorrectedWBCs• Neutropenia/Neutrocytosis• Lymphopenia/Lymphocytosis
• Count100WBCsofOilImmersion(100x)Examination• Reportas%ofWBCs• Induringcount100WBCs,howmanyNRBCsarefound• CalculationofcorrectedWBCs(cells/µL)
100+numberofNRBCsinduringcount100WBCs
WBCcountx100CorrectedWBCs(cells/µL)=
Blood smear reportSTUDENT NAME ________________________________STUDENT ID________________
SPECIMEN CODE______________
RBC morphology:
Color
¨ Normochromia
¨ Hypochromia
¨ HyperchromiaSize
¨ Normocyte
¨ Microcyte
¨ Macrocyte
Shape, give score of grading
¨ Biconcave…………………..…………………
¨ Target cell…………………..…………………
¨ Tear drop…………................…………………
¨ Echinocyte……………….....…………………
¨ Ovalocyte……………….......…………………
¨ Fragmented cells…………....…………………
¨ Other if found (specific)………………………
Platelets:
Number per field: …………. Estimated platelet count (cells/µL): ………………….
¨ Decrease ¨ Adequate ¨ Increase Comments: ……………………………..
Comments: ………………………………………………………………………………….
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: ………………………………………………………………………………….
……………………………………………………………………………………………….
WorksheetfordifferentialWBCandNRBCscount
Column# 1 2 3 4 5 6 7 8 9 10 Total
Neutrophil
Lymphocyte
Monocyte
Eosinophil
Basophil
NRBCs
Howtousethistable:Eachcolumnmustcontain10cells.These10cellsareneutrophilorlymphocyteormonocyteoreosinophilorbasophil.Total10columnx10cellspercolumn=100cellsascalled100WBCs.Induring,100WBCsarecounting.IfNRBCsarefound,noteintotheyellowedcells.
Diagnosisofthalassemiccarriersandthalassemicdiseasesfrom
- completebloodcountand- hemoglobintyping
Thalassemias1.Homozygousb-thalassemia2.b-Thalassemia/Hb E3.Hb Bart,s hydropsfetalis4.Hb Hdisease
ThalassemicDiseases
Hb H diesease Normal
x
P1
F1
a-Thalassemia 1
a-Thalassemia 1
a-Thalassemia 2
a-Thalassemia 2
Normal
x
P1
F1
a-Thalassemia 1
a-Thalassemia 1
a-Thalassemia 1
Hb Bart’s hydropsfetalis
a-Thalassemia 1
x
P1
F1
b-Thalassemia trait HbE trait
bE
bE bE
Normalb-Thalassemia/HbE disease b-Thal. trait HbE trait
Thalassemia Diagnosis
Hematological Data
CS/PS
A2/E
A
F
Bart’s
Hb Type
• Hb A2 (E)
• Hb FQuantitative
• Hb A
• Hb Bart’s• Hb CS/PS
Qualitative
Hb Typing Data
J Hb J MCV J MCH J MCHC J RDW
Interpretationfordiagnosisofthalassemia
MCV (fL) MCH (pg) Hb typing %HbA2(E) %HbF
Normal 88+6 29+2 A2A 2.6+0.4 0.5+0.7
a-Thalassemia 2 trait 83+7 27+3 A2A 2.9+0.3 0.3+0.1
a-Thalassemia 1 trait 68+5 23+5 A2A 2.6+0.3 0.6+0.3
b-Thalassemia trait 64+5 20+3 A2A 5.5+1.3 1.5+1.4
Hb E trait 81+5 26+2 EA 27.8+7.5 1.2+0.9
Homozygous HbE 65+3 20+1 EE 90.2+4.9 4.3+2.7
b-Thalassemia/HbE dis. 67+6 20+3 EF 61.2+13.6 31.1+14.5
HbH disease 67+7 20+3 A2ABart’sH 1.6+1.2 0.6+0.6
Red Cell Indices
Normocyte MicrocyteHb Analysis Hb Analysis
A2 + A A+CS A+E EE A2 + ANormala-thal 2
Thalassemic disease(specified by Hb types)
Hb Constrant Spring
Hb E Determination Hb A2 DeterminationHb E Homozygote
E ~ 25-30% E 21% or less A2 < 3.5%A2 > 3.5%
Hb E traita-thal 2/Hb E
b-thal trait
Hb < 10 g/dl Hb > 10 g/dl Hb < 10 g/dl Hb > 10 g/dl
Hb E withFe deficiency
a-thal 1/Hb E Fe deficiency orFe deficiencyon top of thalassemia trait
a-thal 1 traitHomo a-thal 2
Hb Bart’sHb Bart’s
Casestudy(example)
Mr.Z Mrs.Zx
Riskcouple?
Family1
Hematological DataHb (g/dl) 12.4MCV (fl) 90.5MCH (pg) 29 MCHC (g/dl) 31.5RDW (%) 12.1
A2
A
HbF
HbA
HbA2
Mr.Z
Family1:1. Individualdiagnosis(MaleandFemale)by
give%ofeachHb type,interpretationofHb typinganddiagnosisforthalassemia
2. Givepossibilityofthiscoupletogiveanewbirth.Aretheyariskcoupleforthalassemiadisease?
Hematological DataHb (g/dl) 12.4MCV (fl) 90.5MCH (pg) 29 MCHC (g/dl) 31.5RDW (%) 12.1
A2
A
HbF
HbA
HbA2
Mr.Z
Hemoglobin TypingHb type A2AHb A2 (E) 2.8 %Hb F 0.3 %Hb A 87.5 %
Diagnosis: Normal Hb typing(aa/aa, bN/bN)
Hematological DataHb (g/dl) 9.4MCV (fl) 68.0MCH (pg) 21.5MCHC (g/dl) 31.6RDW (%) 23.0
Hemoglobin TypingHb typeHb A2 (E)Hb FHb A
A2
E
F
HbF HbE
Mrs.Z
Hematological DataHb (g/dl) 9.4MCV (fl) 68.0MCH (pg) 21.5MCHC (g/dl) 31.6RDW (%) 23.0
Hemoglobin TypingHb type EFHb A2 (E) 52.7Hb F 43.2Hb A -
A2
E
F
HbF HbE
Mrs.Z
Diagnosis: b-thalassemia/HbE disease(aa/aa, b0/bE)
b-thal trait Hb E trait b-thal trait Hb E trait
Family 1
b-thal/HbE diseaseNormal Hb typing
Mr. Z Mrs. Z
bE
bEbE
Casestudy(example)
Mr.Z Mrs.Zx
Riskcouple?
Family1
ActivitiesforLab
• Individualword• Choose3specimenbloodsmearwithCode“Nxxx”,“NSxxx”and“Sxxx”foranynumber.SpecifyNo.(xxx)onreport.• Chooseonesetofcasestudy.Thereare4sets.Eachsetcontain2family(4subjects).Reportindividualdiagnosisofthalassemia(4subjects),familypedigreechartandanswerthequestionthattheyareariskcoupleornot.
• Submitreportattheendofclass• Evaluation:• 100%oflabexaminationonL4Anemia&Thalassemia