ICMR-NIIH Practical Guide to Laboratory Immunohematology Shrimati Dharmapal Shetty
INDEX
Page numbers followed by f refer to figure, fc refer to flow chart, and t refer to table.
A
ABO
antibody titer 215
blood group 203
system 208
tube technique for 209
gene 208
titer 217t
weaker variants 208
Absolute neutrophil count 352, 353
Acanthocytes 5
Acetic acid 286
clot lysis method 96
procedure of 97t
Acid citrate dextrose 243, 330, 413
Acid solution 283
Acidic components 4
Acquired immunodeficiency syndrome 236
Acrylamide 469, 474476
Acrylamide stock solution 469
Activated partial thromboplastin time 89, 90, 98, 408
abnormal 91
screening 168
test 92
Acute myeloid leukemia, development of 303
ADAMTS 13 196, 453
activity assay 196
autoantibodies 197
sequencing 198, 199t
Adenosine
deaminase 326, 327
diphosphate 39, 107, 108, 409
triphosphate 33, 40, 403
Adrenaline 107, 409
Adsorption 211
elution test 210, 211t
Agarose 288
gel 57f
electrophoresis 119, 473
Agglutinate 116
cells 226
Agglutination inhibition 218
test 212
Agglutination reaction 208
Agkistrodon contortrix 178
Albumin 223
Aldolase 404
Alloimmunization, prevention of 232
Alpha gene, detection of 64f, 66f
Alpha globin gene
deletions 63f
triplication, detection of 65
Alpha-thalassemia deletions 64t
Alphorids 288
Alternate complement pathway 200
genes of 200t
Amine incorporation assays 98
Ammonia release assay 97
Ammonium
acetate 288, 468
chloride, dissolve 308
persulphate 469
Ampicillin 288
Amplification 263
Amplification-refractory mutation system 58, 58f
technique 59f, 60f, 407
Amplified fragments, resolution of 140
Anemia, classification of 7t
Anisocytes 5
Annealing temperatures 171t, 199t
Anti-A titer 215
normal range for 235t
Anti-B titer 215
normal range for 235t
Antibiotics 288
Antibody 174
defects 340fc
details of 361t
detection 106
iodination of 121
neutralization 212, 218
panels 328
quantitation of 264
titration 322, 323f
Antibody deficiency 319, 321
evaluation of primary 344
primary 344t
Antibody identification 225, 227
from eluate 239
from serum 239
panel cells 229t
Antibody screening 225
cells
commercial 226
typical antigram of 227t
Anticoagulant 407
Anticoagulation with heparin 178
Anticomplement factor H 197
Anticyclic citrullinated peptide antibodies test 273
Anti-D
reagents 203
sera 203
Antifade mounting medium 290
Antifade solution 293
Antiflavocytochrome 361
Antigen 174, 212
antibody precipitation 176
estimation 97
high-frequency 243
positive indicator cells 212
Antiglobulin sera 223
Antiglobulin technique, indirect 228, 233
Antiglobulin test 219, 236
indirect 221, 221f, 223, 234, 239, 241t, 414
technique, cross-match by indirect 241
Anti-H titers 215
Anti-hepatitis B
core antigen 262
surface antigen 263
Antihuman globulin 220, 221
negative 414
reagent 226, 232
sera 219
Antihuman monoclonal anti-fetal hemoglobin antibody 403
Antimitochondrial antibodies 272
Antimycobacterial immunity 367
Antineutrophil cytoplasmic antibodies 268, 270
Antinuclear antibody 268, 270, 272, 274
blot 270
indirect immunofluorescence assay 268
patterns 269, 269t
test 268
Antiperinuclear factor 273
Antiphospholipid antibodies 167
Antisera anti
A 203
B 203
Antiserum 206
Antithrombin 173
activity assay 173
deficiency, types of 173fc
gene 176, 177t
Anti-von Willebrand factor antibody 119
Aplastic anemia 313
investigations 313
Apoptosis assay 382
Arachidonic acid 107, 108, 409
Arginine hot spot mutations, screening for 131
Aspergillus species 321
Atrial fibrillation 188
Autoantibody
against cyclic citrullinated peptides 273
targets 272t
tests for detecting 237
Autoimmune
diseases 267, 268, 272t
disorders 265
hemolytic anemia 207, 236, 237
lymphoproliferative syndrome 375, 382, 383fc
Autoinflammatory disorders 319
Automated electrophoresis technique 46
Autosomal recessive 343, 356
Avidin 290
B
B cells 326, 328, 329, 343, 350, 363f
defects 326, 328, 339
numbers 339
B substance 212
Baby's cord blood, tests for 234
Bacillus-Calmette-Guérin 321
vaccines 365
Bacteria 321
Bacterial artificial chromosome 287f
Bacterial clone deoxyribonucleic acid mini-preparation 287
Bacterial culture work 310
Bacto yeast extract 288
Bactotrypton 288
Basophilic stippling 5
Bernard-Soulier syndrome 110, 110t, 111, 163
Beta globin gene 60f, 62f, 416
Beta thalassemia
diagnosis of 28
major 5, 20f
minor 5
screening of 11f
trait 20f
Beta-mercaptoethanol 290
Bethesda assay graph 105f
Biochemical basis 107
Bisacrolylpiperazine 475
Bleeding
diagnosis of 85
type of 85
Bleeding disorders
genetic diagnosis of 123
laboratory diagnosis of 85
Blocking solution 106, 290
Blood
films, staining of 4, 401
specimen 267
urea nitrogen 236
Blood group 203, 206, 209, 219, 231, 237
A cell 203
antigens, minor 250
B cell 203
O cell 203
positive 205f
red cells 411
results 206t, 245t
Blood sample 366
heparinized 28
Bombay phenotype 208, 209t, 211t, 214, 215, 218, 219, 246
confirmation of 209
Bone marrow 286
aspirate 303
collect 286
examination 352
Borate ethylenediaminetetraacetic acid 469
Bothrops atrox 93
Bovine albumin 290, 361
Bovine serum albumin 121
Branched deoxyribonucleic acid technology 259
Bromophenol 415
Bruton tyrosine kinase expression, detection of 341, 342t
Buffers 406
preparation 412
Bursttest test kit 360
C
C3B/C4B inactivator 198
Calcium
ionophore 347, 380
solution 346, 379
presence of 107
Candida species 321
Capillary electrophoresis 481
Carnoy's fixative 307
CAT technology 226, 228
Cation exchange high-performance liquid chromatography 18, 28
CD119 on monocytes, expression of 365
CD154 expression
ranges 350
reagent for 347t
CD20 expression 340, 343
CD4 lymphopenia 326
CD40
expression 343
gene 343
CD40l
expression 343
gene 343
CD8 lymphopenia 326
Cell
button 217
culture pipette 307
culture, basic requirements for 283
grouping 203, 204, 206, 206t, 209
lysis 470
transfused 208
Cellulose acetate electrophoresis 16, 16f, 402
Central nervous system 196, 272
Centrifuge 285, 287
Centromere pattern 269
Centrosomic nucleoprotein B 272
Charge-coupled device 293
Chloramphenicol 288
Chloroform 468
Chorionic villus sample 132
Chromogenic assay 101
activity by 173
Chromophoric group 101
Chromosomal breakage investigation 307
Chromosomal instability syndromes 283
Chromosome 294f
analysis 284
denaturation 295
preparation 286
Chronic myeloid leukemia 297
diagnosis of 297
monitoring of 297
Citrate phosphate dextrose 243, 413
Classical bethesda assay 104
Clauss assay, modified 94
Coagulation cascade 88f
Coated autoantibodies, elution of 239
Coating buffer 106
Colchicine stock solution 286
Cold
agglutinin syndrome 236
antibodies, titration for 239
centrifuge 315
hem-agglutinin disease 236
Collagen 107, 409
binding assay 86, 117
Column agglutination technique, cross-match by 223
Combined immunodeficiency
severe 326, 327, 331f
workup
protocols for severe 328
severe 326
Commercial panel cells 228
Complement control protein 197
Complement deficiency 319, 321, 386, 386fc
Complement factor
B 198
H 197
I 198
Complementary deoxyribonucleic acid 259, 299
conversion procedure 298
master mix components 298t
preparation for 299t
thermal cycler conditions for 299t
Complementary test 147f
Complete blood count 3, 6, 36
Complex translocation
evaluation of 294f
spectral karyotyping of 296f
Conformation sensitive gel electrophoresis 124, 474
gel composition 476
Congenital neutropenia, severe 353
Connective tissue disease, mixed 271, 272
Consensus repeats, short 197
Conventional staining 286
Coomassie stain 405
Coombs control cells 414, 414t
Copy number status dosage quotient 482
Counter stain 293, 402
Covalent reverse dot-blot hybridization 52, 56f
technique 405
Crithidia luciliae 269, 270
Cross-matching procedures 222
Cross-reacting material 102
Cryptococcus neoformans 321
Cryptosporidium parvum 321
Cyanine 289
Cycle sequencing program 81t, 477
Cyclic citrullinated peptide 279
CYP2C9*2
genotype interpretation 191f
polymorphism 190t
CYP2C9*3
genotype interpretation 192f
polymorphism 190, 191, 191t, 192
CYP4F2:V433M polymorphism 195t, 196
Cysteine 411
activated papain 415
Cytochrome B-245 alpha 355
Cytofix 369
Cytogenetics 281, 460
conventional 303
laboratory setup 283
Cytomegalovirus 321
Cytometer set-up and tracking 323
Cytoperm 369
Cytoplasm 4, 270
Cytoplasmic antineutrophil cytoplasmic antibodies 270
D
Dde1 alleles, resolution of 153f
Deionized water 43, 475
Delta globin gene
defect 67f
mutations, detection of 67f
Delta thalassemia, detection of 66
Denaturation solution 288, 295
Denaturing gradient gel electrophoresis 138
Dendritic cells 350
Deoxyadenosine triphosphate 289
Deoxycytidine triphosphate 289, 290
Deoxyguanosine triphosphate 289
Deoxyribonuclease 289
Deoxyribonucleic acid 52
antibodies, anti-double stranded 269
denaturation 480
extraction 314, 470
polymerase 289
precipitation of 470
quality of 471
quantity of 471
sequencing 73f, 302, 312, 476
direct 148, 155
electropherogram 67f
exon-wise 75
of beta-globin gene 60
protocol 388t
reaction 81t
spectral karyotyping 294
Deoxyribonucleotide triphosphate 289
Deproteinization 470
Dermatomyositis 271
Destaining solution 402
Detection buffer 290
Diagnostic and complementary tests 148t
Diamidino-2-phenylindole 291
Dichlorophenolindophenol
dye test 404
test 36
Diepoxybutane 306
Diethylaminoethyl 234
Digested fragments
interpretation for 183t
resolution of 138, 139, 143, 153, 181, 182, 184, 189
Dihydrorhodamine 353, 355, 358
assay 355, 357, 359f
Dilute Russel viper venom time test 167, 169
Dimension gels, first 176
Dimethyl sulfoxide 347, 356, 379, 380
Dimorphic population 5
Dipotassium hydrogen phosphate 401
Direct antiglobulin test 207, 219, 220f, 237
Direct coombs test 34
Direct thrombin inhibitors 102
Disodium
hydrogen phosphate 412, 415
phosphate 401
Disposable syringes with needles 285
Disseminated intravascular coagulation 170
Distilled water 3840
Dithiothreitol 45, 415
reagent, preparation of 415
Dolichos biflorus 203, 410, 411
Donors blood sample 242
Dot plots 339f
Dot-blot hybridization, reverse 52f
Double negative T cells 382, 383
evaluation of 384t
Double negative T lymophocytes, flow cytometric evaluation of 385f
Dual fusion translocation probe 297f
Dual-platform method 328, 329
Duffy blood group antigens, genotyping of 254f
Dysfibrinogenemia 93
E
Eagle's medium 310
Dulbecco's modified 310
Earl's medium 295
Electropherogram 246f
Electrophoresis 120, 121
apparatus 287
grade 119
specifications 481
Electrophoretic buffer 120
Elevated serum globulin 236
Elliptocytes 5
Elution 211
solution 402
volume, calculation of 17
Embden-Meyerhof-Parnas pathway 34
Endothelial protein C receptor 184, 185t
Enteric flora 321
Enteroviruses 321
Enzyme 223, 241t
immunoassay 269
method 239
technique 223, 228, 233
titer cross-match by 241
Enzyme-linked immunosorbent assay 95, 97, 102, 106, 112, 262, 263, 272, 382
Eosin-5-maleimide 404
fluorescent dye 44
Epinephrine 107, 409
Epithelial cells 212
Epstein-Barr virus 310, 321
Erythrocyte 210
lysis buffer 308
Escherichia coli 310
Ethanol 288, 402
series 292, 294
washing 477
Ethidium bromide 469, 472
Ethylene glycol 475, 476
Ethylenediaminetetraacetic acid 38, 47, 203, 204, 210, 259, 287, 288, 344, 353
Exon 430
Exon14 bsl 1 140
Extragenic marker 140
F
F cell estimation 403
F-10 nutrient mixture 284
Factor assays, different modes of 100t
Factor deficient plasma 408
Factor II gene 135, 135f
Factor IX
antigen estimation 102
gene 152, 152f
inhibitor screening assay 103
Factor V gene 136, 136f
Factor V Leiden
genotypes 179f
mutation 178, 179t
interpretation for 180t
Factor VII gene 137, 137f
Factor VIII
antigen estimation 102
gene 138
inhibitor screening assay 103
Factor X gene 157
Factor XI gene 157, 158f
Factor XII gene 159, 159f
Factor XIII
estimation 95
gene 160, 160f
Familial hemophagocytic lymphohistiocytosis 375, 377
Family history and pedigree 85
FANC genes mutation analysis 311
FANCD2 immunoblot patterns 309t
FANCD2 monoubiquitination detection 308, 309f
Fanconi anemia
basic diagnostic method for 306
molecular investigations for 310
workup 306
F-cell estimation 32
Ferric chloride 402
Fetal blood analysis 28
Fetal bovine serum 284, 307
Fetal calf serum 310
Fetal cell
estimation by flow cytometry 32f
staining 11, 12f
Kleihauer-Betke method of 402
Fetal D antigen 230f
Fetal hemoglobin
acid-resistant 11
hereditary persistence of 3, 68, 71
Fibrinogen
concentration 95
estimation of 94
genes 134, 134f
Flow cytometer-based assays 322
Flow cytometry 32, 42, 341, 342t, 343, 355, 357, 360, 361, 364, 403
analysis 44, 111f, 335f, 404
data acquisition and analysis 333, 335, 337, 342, 345, 351, 354, 359, 362, 378, 385
evaluation 337f, 343f, 385f
osmotic fragility 42
Fluorescein isothiocyanate 268, 322
Fluorescence activated cell sorting lyse 358
solution 335, 336, 369, 384
Fluorescence dye 404
Fluorescence histogram 44f
Fluorescence in situ hybridization 283, 287, 291, 292f, 294f, 297, 297f, 303
rapid 295
technique 297
Fluorescence resonance energy transfer 196
Fluorescent compounds 322, 324
Fluorescent fragments, short 306f
Fluorescent in situ hybridization, slides for 291
Fluorescent microscope using different filters 293
Fluorescent-labeled antibodies 110t
Fluorochrome 289, 322
selection 322
Formaldehyde 341, 360, 361
fixed platelets 116
Formamide 292, 295, 476
Forward scatter 342, 354
characteristics 43
Fragment separation 481
Frozen plasma samples, controlled thawing of 87
Frozen serum samples, controlled thawing of 267
Fucosyltransferase
2 deletion, detection of 248
gene 246, 249f
analysis 250f
FXIII deficiency, detects mild-to-moderate 96
G
Gamma chain, detection of 336, 336t
Gamma-activated
factor 367
sequence 367
Gastrointestinal diseases 321
Gastrointestinal tract 321
G-banded metaphase 294f
Gel
card, agglutination reaction in 224f
electropherogram 153f, 186f, 193f195f
extraction 477
loading dye 469, 473
staining dye 474
Gene 416, 430, 458, 460, 466
encoding
ankyrin-1 34
GPIB/IX/V complex 163f
transfection 310
General molecular biology protocols 468
Genesplicer 484
Genetic diseases, diagnosis of 283
Genomic deoxyribonucleic acid
digestion of 145t
with Bcl1, digestion of 144
Genotype 59f, 144, 184
different 50t
interpretation of different 57t, 71t
techniques 250
Giardia lamblia 321
Giemsa stain 286, 287
Giemsa-banding technique using trypsin 287
Glacial acetic acid 285
Glanzmann thrombasthenia 110, 110f, 161
diagnosis of 110t
Glass Pasteur pipettes 307
Glasswares, preparation of 283
Globin gene promoters 71
Glomerulonephritis 270
Glucose 403
phosphate isomerase 34
activity 39
assay reaction composition 40t
deficiency 78f
Glucose-6-phosphate 38
dehydrogenase 34, 36, 40, 74
activity test 37
activity, calculation of 38
assay reaction composition 38t
deficiency 73, 404
detection of 75f, 76f
gene 74t
screening for 36
Glucose-tris-ethylenediaminetetraacetic acid lysis buffer 288
Glutaraldehyde 403
3-phosphate dehydrogenase 299
Glycerol-citrate
buffer 415
preservative 413
Glycerolization, procedure of 413
Glycoproteins 110
GP IIB gene 161
GP1BB gene 163
GP9 gene 163
GPIBA gene 163
GPIBB gene 163
GPIIB gene 161
GPIIIA gene 161, 163t
Granular cytoplasmic staining, diffuse 270
Granule release assay 375, 378, 380t, 382t
Granulomatous disease, chronic 352, 354, 355, 360
H
Haemophilus influenzae 321
Heat denaturation technique 14
Heat elution 237
Heat stability test 14f
Hematocrit 7
Hematology 61t
Hematopoietic stem cell transplantation 319
Hematoxylin 402
Hemoglobin 7, 16, 36
A2 28
quantitation of 17
adult 16
analysis 18
C 3
D 3, 48, 51f
disease 6
mutation, detection of 49t
detection of unstable 402
disorders of 3
E 3, 16
beta thalassemia 24f
homozygous 24f
minor 6
trait 23f
electrophoresis 4
H 16
disease 3, 6, 21f
M, spectrophotometric detection of 31
N 31
S 48, 51f
beta-thalassemia 6
confirmation of 48
heterozygous cord blood sample 30f
solubility test for 10f
tests for unstable 14
unstable 14f
variant, unstable 15
Hemoglobinopathy 1, 401, 416
detection of 48
laboratory investigation of 3
Hemoglobins, abnormal 17
Hemolysate, preparation of 13, 35
Hemolysis 217
Hemolytic disease
cause of 230
of newborn 165, 203, 230, 234
Hemolytic transfusion reactions, investigations of 242
Hemolytic uremic syndrome 196
atypical 196
Hemophagocytic lymphohistiocytosis 375, 376fc, 387
workup 376
evaluation of 380t
Hemophilia
A 138, 144
genetic diagnosis of severe 151fc
mild 102
B 152
genetic diagnosis of 157fc
Hemostasis 83, 407, 430
Heparin 330
binding site 176
presence of 93
vacutainers 284
Hepatitis
B
core antigen 262
surface antigen 263, 264
B E-antigen 264
enzyme-linked immunosorbent assay 264
B virus 259, 263
deoxyribonucleic acid quantitation 263
C virus 259, 260
viral load 259
Hereditary spherocytosis 43f, 44f
Heteroduplex 476
formation, sample for 476
Heterozygous alpha thalassemia 5
Hexokinase activity 40
Hexose monophosphate pathway 34
Hha1 alleles, resolution of 154f
High performance liquid chromatography 36
Histocompatibility complex, major 326, 327
Histoplasmosis capsulatum 321
Homogeneous pattern 269
Homozygous
deletion, large 311f
genotypes 48
Horseradish
peroxidase 262
peroxide 117
Hot air oven 283
Human albumin 356
Human chromosome preparation 284
Human globulins 219
Human immunodeficiency virus
type 1 and type 2 259
viral load 262
Human interleukin-7 receptor subunit alpha 334
Human leukocyte antigen 326, 327
Human plasma 263
Human platelet antigen 165
5 alleles 165f, 166f
genotyping 165
Human splicing finder 484
Hybridization 292, 293, 406
chamber 292
procedure 55
reaction 480
Hydrochloric acid 197
Hydrochloride 39
Hyper immunoglobulin M
defect 387
protocols for 343
syndrome 343
Hypochromia 5
Hypotonic solution 286
I
Imidazole buffered saline, preparation of 407
Immune
dysregulation 375
diseases of 319
Immunodeficiency
combined 321, 326
common variable 339, 340
disorders
genotyping of primary 387
molecular analysis of primary 387
primary 317, 319, 320, 321t, 322, 365, 466
screening, primary 325
syndromes 319
Immunoelectrophoresis 175
Immunoglobulin
dissociation of 238
G 220, 221, 233, 234
antibody 233t
coated red cells, preparation of 414
levels, low 326
In vitro incubation 9f
Incubated mix reading 103
Indirect immunofluorescence
assay 268
technique 268
Inflammation, chronic 352
Inhibition 219
Inhibitor screen test 103
In-house panel cells 227
preparation of 412
Innate immunity 319
defects 364
Interferon gamma receptor 364, 366t, 368, 369t
on monocytes, expression of 368
Interleukin 12 receptor beta 1 370, 371t
Interleukin 7 receptor subunit alpha, detection of 334, 334t
International normalized ratio 89
International System for Chromosome Nomenclature 287
Interstitial lung disease 277
Intracellular phospho-stat assay 332, 332t
Intracellular staining protocol 362
Intracranial hemorrhage, cause of 165
Intravenous blood 267
Intrinsic immunity defects 364
Intron 1 GT repeat marker 140, 142t
Intron 1 inversion analysis 148, 149t
Intron 22 inversion analysis 144
Intron hind III
reaction mix for 140t
re-digestion for 140t
Inverse protein C resistance technique 144
ISO-osmotic phosphate buffer 401
Isopropanol 288
test 15
Isopropanol-Tris buffer 402
ITG3A gene 161f
ITGA2B gene 161f
J
JAK2 V617F mutation 188f
detection of 187
K
Kahn tubes 232
Kanamycin 288
Karyotype preparation 287
Kell blood group antigens, genotyping of 255f
Kidd blood group antigens, genotyping of 254f
Kidneys 270
Kinetoplast 269
Kleihauer-Betke method 11, 12f, 402
Kleihauer-Betke test 11
L
Lactate dehydrogenase 39, 403
Laemmli buffer 308
Laminar flow 283, 285
L-asparaginase therapy 170
Laurell immunoelectrophoresis 114, 115f
Lectins 410, 411
dolichos biflorus 410f
preparation of 411
Leishman stain 4, 5f, 401
Leptocytes 5
Leukocyte adhesion
defect 387
deficiency 352, 353
protocols for 352
Ligase buffer
A 479
B 479
Ligase-65 479
Ligation reaction 480
Linkage analysis 138
Liquid chromatography chromatogram, high-performance 19f, 20f27f, 29f, 30f
Listeria monocytogenes 321
Liver 321
disease 90
dysfunction 170
Lower respiratory tract 321
Lung
abscesses 321
vasculitis of small arteries of 270
Lupus anticoagulant 167, 168
testing 87
Lupus nephritis 275
Luria-Bertani medium 288
Lymphadenitis 321
Lymphoblastoid cell lines
maintenance of 310
preparation of 310
Lymphocyte 343
cell culture, maintenance of 310
function-associated antigen-1 352
identification of 330f
preparation and culture procedure 308
subset analysis 325, 326, 328, 340, 364, 382
subset report 331f
Lysate preparation 313
Lyse wash-stain-wash 325
M
Macrocytes 5
Macrophage-activation syndrome 375
Macrothrombocytopenia 163
Magnesium chloride 38
Maternal serum antibody 232
Mature human erythrocytes 33
Maxentscan 484
Mean channel fluorescence 44
Mean corpuscular
hemoglobin 7
concentration 6, 7
volume 7
Median fluorescence intensity 323, 334
Membrane protein defects, identification of 46
Membrane-Biodyne C transfer membrane 406
Memory B cells 343, 344
pattern of 346f
Memory cells 337
Memory T cells 337, 338t
Mendelian inheritance 319
Mendelian susceptibility 353, 364, 365
Meningitis 321
Mepacrine 112
Metaphase plate 296f
Methanol 285, 286
Methemoglobin 30
absorption maxima of 31f
measurement of 30, 402
Methylenetetrahydrofolate reductase 167
A1298C polymorphism 182
C677T polymorphism 182, 183f
enzyme, activity of 181
gene polymorphism 181, 182t
Micro pipettes 292
Microangiopathic hemolytic anemia 196
Microcentrifuge tubes 292
Microcytes 5
Mitomycin C 306
Mitotic index 287
Molecular biology
protocols 48
reagents, common 468
Molecular study 343, 382
Monoclonal antibody 348, 377, 381
panels
four-color 329
three-color 329
Monoclonal immunoglobulin M 203
Monocytes 340, 350, 363f
Mononuclear cell separation 310
Moraxella catarrhalis 321
Multiple ligation-dependent probe amplification, advantages of 482
Multiple vitamin K dependent coagulation factor deficiency 93
Multiplex ligation dependent probe amplification 131, 131f, 311, 478
buffer 479
kit component 479
reaction, thermocycler program for 480
technique 131
Mutation 416
analysis in genes 200
direct detection of 144
screening strategy 124
Mycobacteria 321
Mycobacterial disease 353, 364, 365, 365fc
workup 364, 366t, 369t, 371t, 373t
Mycobacterium tuberculosis 365
Mycoplasma pneumoniae 321
Myelodysplastic syndrome 303
diagnostic workup of 304fc
Myocardial infarction 188
N
Naïve B cell 343, 344
Naïve cells 337, 338t, 343
Naïve T cells 337
Naked eye single tube red cell osmotic fragility test 10, 11f
Nasal passages 270
Natural anticoagulants protein C 167
Natural killer cell 326329, 336, 370, 378
function 375
Neisseria meningitides 321
Neonatal alloimmune thrombocytopenia 165
Nephelometry-based assays 343
Nestroft test 401
Neutrophils 350, 357f
cytosolic factor-2 355
gating of 363f
New methylene blue stain 401
Nick translation
buffer 290
method 289
Nicotinamide adenine dinucleotide 39, 79
reduced 34
Nicotinamide adenine dinucleotide phosphate 34, 38, 40, 98, 403
oxidase 354, 355
oxidase component
evaluation of 360
expression, detection of 361
Nijmegen modified Bethesda assay 104, 104f
Nitroblue tetrazolium 52, 352, 353, 355, 356, 365
blood test 325, 355, 356, 364
concentration of 356
plain reagent, concentration of 356
Nitroblue tetrazolium test
interpretation of 357
microscopic examination of 357f
negative 357
positive 357
Nonfunctional receptor 367f
Nonsecretor saliva 213
Nontuberculous environmental mycobacteria 365
Normal cord blood sample 29f
Normal healthy control 109f
Normal pooled plasma
dilutions 174fc
preparation of 408
Nucleic acid 470
sequence-based amplification 259
Nucleolar pattern 269
Nucleotide deletion 73f
Nucleotide polymorphisms, single 482
Nucleotide replacement 261
O
O cells 225
O group
cells 225
red cells 411
O pooled cells 232
O RhD
negative 234
positive cells 234
Octanol 468
Oligo probes, blotting of 406
Oligonucleotide probes 406
Omenn syndrome 326
Optical density 113
Osmotic fragility test 42
Otitis 321
Owren's barbiturate
buffer 407
saline 98, 408
Owren's buffer 98
Oxidative burst function 357f
Oxidized glutathione 404
P
Packed cell volume 6, 7
Panel cells 412
Papain 411
cysteine 232
Para-Bombay
blood group 217t
phenotype 208, 209t, 211t, 214, 215, 219
Parainfluenza type 3 321
Partial autosomal dominant form-over expression 367f
Partial thromboplastin 90
Pasteur pipette 285, 286
Patassium chloride 39
Perforin expression 376
analysis 379f
Perforin for hemophagocytic lymphohistiocytosis workup, evaluation of 377t
Perinuclear antineutrophil cytoplasmic antibody 270
Perinuclear staining 270
Peripheral blood 262
smear 5f, 9f
Peripheral venous blood samples 322
Phagocyte
destroy bacteria 356
number and function, congenital defects of 319
Phagocytic defects 321, 352, 353fc
Phenol 468
Phenol-chloroform method 470
Philadelphia chromosome A 294f
Phorbol myristate acetate 356, 358
reagent 356
Phorbol-12 myristate-13 acetate 356, 358, 380
Phosphate buffer 401, 402
saline 47, 268, 308, 361, 403, 415
Phospho signal transducer and activator of transcription
assay, detection of 368t
on monocytes, expression of 366, 372
Phosphoenolpyruvate 39, 403
Phospholipids 90
Phosphorylated signal transducer 364
Phytohemagglutinin 284, 371
Plant lectin 203
Plasma
samples 120
specific factor deficient 98
Plasminogen activator
inhibitor 1 activity 185
tissue-type 185
Platelet 111f
aggregation
basic pathways of 108
graph 109f
reagents 409
tests 107
dense granules 112
fresh-washed 116
poor plasma 87
receptor studies 110
rich plasma 409
Pneumocystis jiroveci 321
Pneumonia 321
P-nitro aniline 101
Poikilocytes 5
Polyacrylamide gel 76f
electrophoresis 165f, 166f, 473
preparation 469
technique 45
Polyacrylamide stock solution 474
Polychromasia 5
Polychromic staining solution 4
Polyclonal antibody 403
Polygalacturonic acid 404
Polymerase chain reaction 49, 58, 74, 81, 123, 172t, 259, 300, 305, 472
allele-specific 187, 187t
amplification 146t, 151t, 312
checking for 55, 252
protocols 132
components 138, 299t
composition 245t
conditions 314t
first 300t
for hematology 61t
master mix preparation 300t, 301t, 316t
composition 299, 387t
mixture 65t, 72t
multiplex 64f, 66f
program 61, 61t, 249t, 299
protocol 299
seminested 300
purification 301, 477
quantitative 303, 304, 314
reaction 247, 481
reagent
composition 125f, 133, 149t
real-time 259, 263, 299t
second 301t
thermal cycler conditions
for first 300t
for second 301t
for sequencing 302t
transcription
analysis, reverse 298f
reverse 313
test, reverse 297
Polymorphic alleles 154f156f
Polymorphic markers 152f
Polymorphic regions 152t
Polymorphism 167
analysis 48
heterozygous for 138
phenotyping 483
strongly-associated 188
weakly-associated 188
Polymyositis 272
Polyphen 483
Polypoidal choroidal vasculopathy 198
Polysaccharide 340
Ponceau S stain 402
Pooled cells, preparation of 410
Posthybridization washing 291293
solution 292
Postsequencing product cleanup 302
Postsequencing reaction cleanup 477
Potassium
bicarbonate 308
chloride 285
cyanide 402
dihydrogen phosphate 401, 412
ferricyanide 402, 403
Power SYBR master mix 316
Precipitating rabbit antiserum 174
Precipitin tubes 232
Pre-warmed hypotonic solution 286
Probes in membrane 53f
Proliferating cell nuclear antigen 272
Promoter polymorphism 186t
genotypes 186f
Promyelocyte arrest 352
Protamine sulfate neutralization 93
Protein 340, 470
analysis 483
and denaturation, dilution of labeled 46
C
activated 177
activity 170
deficiency 170
gene 170, 170f, 171t
resistance, activated 177
S
activity 171
gene 172, 172f, 172t
Proteinase K 468
Proteolytic enzymes 411
Prothrombin G20210A polymorphism, alleles of 181f
Prothrombin gene
G20210A polymorphism 180
polymorphism 180t, 181t
Prothrombin time 86, 88, 89
abnormal 91
Protozoa 321
Pseudomonas aeruginosa 321
Pulmonary arterial hypertension 277
Purine nucleoside phosphorylase 326
Pyrimidine 5'-nucleotidase activity
detection of 404
screening test for 41
Pyruvate kinase
activity 38
assay reaction composition 39t
deficiency, molecular characterization of 80f
Q
Qualitative deficiency 173
Quantitative deficiency 173
R
Radiation sensitivity test 326
Radiolabeling antibodies 121
Rare blood 243
group 243
Rare donor cells, preservation of 243
Rare phenotype 243
Re mix for exon 14 BSL1 142t
Reaction composition 38, 39
Reaction mixture preparation 314t
Reactive oxygen species 360
Reagent 332
and solutions 346, 379
pooled cells 410
preparation of 286, 290, 335, 336, 344, 354, 358
red cell 243
panels 412
Receiver operating characteristic 273
Reciprocal translocation, double 297f
Red blood cell 7, 36, 42, 220, 221, 230f, 239, 325, 326
distribution 7
enzymopathy, diagnosis of 36fc
lysis buffer 405
membranopathy, diagnosis of 36fc
parameters 7t
Red cell
agglutination of 205
disorders 34
enzyme assay 35
reagent for 403
enzymopathies 1, 33
detection of 73
membrane 34
disorders 42
ghost, preparation of 46, 47fc
membrane protein
analysis of 46f, 48f
defects 48f
membrane protein deficiency 44
flow cytometric analysis of 404
membranopathies 1, 33, 73
pellet 225
pooled 206
preparation of 243, 413
preservation of 243, 415
protein defects, identification of 45
suspension, preparation of 42
Renal tubular acidosis, distal 76
Representative gel 179f
Reptilase time 93
reagent 94
Respiratory syncytial virus 321
Respiratory tract 321
Restriction enzyme 74t
digestion 50, 75
reaction mix 50t
Restriction fragment length polymorphism 247t
technique 51f
Reticulocyte
body 8f
count 7, 401
Retroviral gene transfection 310
Retroviral production 310
Rh antibody, titration of 232, 233t, 234t
Rh genotyping 231
Rh hemolytic disease 230
Rh phenotypes and genotypes 231t
Rh zygosity 231
RhD antigen 230
RhD gene 251
RhD genotyping test 256f
RhD grouping 207, 231
RhD negative
mother 230
phenotype donors 253f
RhD positive phenotype donors 253f
Rheumatic factor detection and quantification 272
Rheumatoid arthritis 272, 279
Rheumatoid factor 272
Ribonuclease 288
Ribonucleic acid 297
extraction 297
Ribonucleoprotein 272
Ribosomal phosphoprotein 272
Ristocetin 409
absent aggregation with 163
cofactor assay 116
Ristocetin-induced platelet aggregation 112
Rocket immunoelectrophoresis 174
Room temperature 4
Running buffer 119
Running gel 120
Russell viper venom 169
S
Saline 241t
normal 232
technique 223, 227, 232
cross-match by 240
titer 235
Saliva
amount of 235
collection of 213
Salmonella
infections 370
typhi 321
Sample buffer 119
Sample dilution buffer 106
Sample preparation 120
Saponin 401
Scleroderma 271
Screening cells 226
Screening coagulation
results, interpretation of 93t
tests 88, 88f
Screening techniques 95
Second dimension gels 176
Secretor status 212
test 214t
Semi-log graph paper 99f
Sensitizing antibody 219
Sepsis 321
Serological diagnosis 203
Serum
amount of 235
grouping 203, 204, 206, 206t, 209
inhibition test 218, 219t
sample 232
Serum immunoglobulin estimation immunoglobulin
A 325
E 325
G 325, 343
M 325
Shaker incubator 287
Sickle cell 9f
anemia 6
disease 28
hemoglobin 3, 16
beta thalassemia 23f
D disease 26f
E disease 25f
homozygous 22f
solubility test for 10
trait 22f
Sickling test 401
for sickle cell hemoglobin 9
Signal transducer 372
Signal-to-noise ratio 323f, 334
Siliconized glass tubes 104
Single platform method 328, 329
Single strand conformation polymorphism 475
Single-chain precursor 198
Single-nucleotide polymorphism 165
Single-platform method, analysis for 331
Sinusitis 321
Sjögren's syndrome 278
Skin infections 321
Slides, rapid ageing of 291
Sodium
acetate 288, 469
bicarbonate buffer 405
chloride 401
dihydrogen phosphate 401, 415
metabisulfite 9f
phosphate 405
Sodium dodecyl sulfate 45, 119, 288
polyacrylamide gel
electrophoresis 36, 47
reagents, preparation of 405
Solid surface antigen 106
Solubility test 401
Sorensen's buffer 286, 287
Speckled pattern 269
Spectral karyotyping reagents 294
Spectral overlap 324
Spherocytes 5
Splice site prediction softwares 483
Stabilizing solution 403
Stacking buffer 119
Stacking gel 120
Stain-lyse-no wash 325
Stain-lyse-wash 325
Staphylococcus aureus 321
Sterilization procedures 284
Stimulate peripheral blood
natural killer cells 380
T-cells 347
Stock Laemmli buffer 405
Stock solution 119, 358, 401, 405, 407
Stomatocytes 5
Streptavidin-peroxidase 198
Streptococcus pneumoniae 321
Stroke 188
Substrate solution 106
Sulfuric acid 197
Synthetic chromomeric substrate 173
Systemic bacterial infections 321
Systemic lupus erythematosus 270, 272, 275, 276
disease activity index 276
Systemic sclerosis 277, 278
Systemic viral infections gastroenteritis 321
T
T-cell 328, 329, 336, 337, 363f
defects 326, 328
receptor 383
T-cytotoxic cells 328, 329, 339f
T-helper cells 328
Tag antibody 113
T- and B-cell
defects, combined 327fc
immunodeficiency, combined 319, 326
Tandem repeats, variable number of 132
TaqMan assay principle 260
Target cells 5
Tc cells 378
T-cell counts
low 326
normal 326
Telomerase activity
analysis 314t
assay 313
Telomerase detection kit, quantitative 313
Telomerase reaction 314
Telomere length
detection 316t
real time thermal cycler conditions for 316t
measurement 314, 315
Telomere standard, sequential dilution of 316t
Test plasma, dilution of 105
Tetramethylbenzidine 122
Tetramethylenediamine 273
Thalassemia 69f
major cord blood sample 29f
T-helper cells 329, 339f
Thermal cycling conditions 54t, 57t, 59t, 66t, 68t, 70t, 72t, 74t, 81t, 133t, 135t137t, 159t161t, 164t, 182t, 245t, 255t, 299
for intron hind III 140t
Thermal cycling parameters 200t
Thrombin
excess of 173
low concentrations of 108
time 86, 92, 93
Thrombocytopenia 196
Thrombophilia
investigations 167
markers, common 167
Thromboplastin
dilution test, modified 100
time 86
Thrombosis 83, 407, 430
Thrombotic microangiopathy 196
Thrombotic thrombocytopenic purpura 196
Thymic epithelial cells 350
Thymic stromal lymphopoietin 334
Tissue culture media, preparation of 284
Toxoplasma gondii 321
Transcription 4 activator of 372
Transcription-mediated amplification 259
Transfection 311
Transfusion medicine 201, 409, 458
genotyping in 244
Transfusion transmitted disorders 257
Triose-phosphate isomerase 404
Tripotassium citrate 415
Triradial formation 308
Tris-acetate-ethylenediaminetetraacetic acid 469
Trisodium citrate 407
Triton X 341, 361, 402, 403
solution 341, 361
Trypsin 287
treatment 295
Tube technique 203, 226, 227, 232
cross-match by 223
Tuberculosis 365
Tyrosine kinase
domain mutation
detection of 300
study 300
inhibitor 297
U
Ulex europaeus 410, 410f, 411
Ultrapure formamide 475
Ultraviolet light 285
source 283
Unstable buffer 402
Upper respiratory tract 321
Urea clot lysis technique 95, 96t
Urinary tract problems 321
Urokinase plasminogen activator 185
V
Venous thromboembolism 177
Viral ribonucleic acid extraction 262
Viruses 321
Vitamin
K
deficiency 170
dependent factors 101
von Willebrand disease 109f, 112, 131f, 132f, 409
diagnosis of 112, 123
von Willebrand factor 85, 86, 116, 117, 119, 121, 123, 123f, 124f, 132
antigen 112, 115f
cleaving protease 196
collagen binding 118f
determination of 114
gene 123
hot spot mutations of 125t
multimer analysis 119
Vortex tubes 347
W
Warfarin sensitive gene polymorphisms 188
Warfarin therapy, indications for 188
Warm antibodies 239
titration for 239
Wash buffer 106, 341, 360, 361
Washing solution 292
Water bath 285, 292, 315
Wegener's granulomatosis 270
White blood cell 6, 7, 47, 336
Witebsky's partial neutralization technique 234
Working sample buffer 120
World Health Organization 89
X
X-linked agammaglobulinemia 339, 340, 387
diagnosis 340, 341t
workup 340
Xylene cyanol 415
Z
ZZAP
reagent 237
treatment 237, 415
reagent for 415
×
Chapter Notes

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