ABO TUBE TEST 20S centrifuge

1+ : turbid
Gel Technology Dextran-Acrylamide Gel
Yves Lapierre

Principle: hemagglutination
Incubation 15 mins
10 mins centri
Standardization of equipments: advantage
Disadvantage: need special equipment
2-3 days: stable reactions
4+ solid band top
3+ predominant
Negative: bottom pellet rbc compatible
Tube centri 20 s
Microtiter well Solid-Phase Technology
2 mins incubation
Chromosome 46 chromosome
23 pairs

2 sex chromosome/ 1 pair

22 autosome pair
Father of Genetics Gregor Mendel
Sweet peas
1st law inheritance: Independent segregation
2nd law: Law of independent assortment
Simplest mutation Point mutation
- Substitution, insertion, deletion
Missense point Codon change
POPE INNOCENT VII (1942) First blood transfusion
ABO Blood grouping Nobel prize winner Karl Landsteiner
Specificity of Serologic Reactions (SSR)
Braxton Hicks Sodium phosphate
Edwards Lindemann Vein to vein
Lewisohn Minimum amount of citrate for anticoagulant
Hustin Sodium citrate
Dr. Charles Drew American Red Cross
1943 ACD Loutit and Mollison
CPD 1957 Gibson
In vivo sensitization
Warm haemolytic anemia Autoimmune haemolytic anemia
Antihuman globulin test/ Coomb’s test Maternal antibody coating fetal RBCs
HDN Specimen: infant’s rbc
DAT EDTA (Pink)
First step DAT: washing
Indirect AHG Test In vitro (In tube/glass) sensitization
 - Weak D, K, Fy (RBC antigen detection)
Enhancement medium AHG Test LISS, PEG, POLYBRENE, AND ALBUMIN
LISS decrease incubation period (10-15 mins)
Albumin (30 mins incubation)
Major Crossmatch Critical crossmatch
ISBT name
001 – ABO
004 – Rh
030 – Rh – associated glycoprotein
4 diff forms of H antigens - Unbranched straight chain (H1 H2, H3, H4)
Anti H / O cells – Ulex europaeus No agglutination with Bombay individuals (No H
Anti A1 : Dolichos biflorus antigens)
Anti B : Bandeiraea simplicifolia
ABO Genes 5 different phenotypes
Bombay Phenotypes (Oh) Hh genotype
No H antigens formed; no A or B antigens formed
Yellow B
Blue Angel
ABO discrepancies
Group 1
Group 2: missing Ag forward grouping (least
frequently encountered)
Group III Rouleaux (phase in Antibody screen: AHG)
Plasma factors
Elevated fibrinogen
Dextran
Wharton’s jelly in cord blood samples
Group IV Discrepancies Anti – I
Oh bommbay Anti A, Anti-B, Anti O
All reactions: rouleaux
Cold antibody (anti-I )

Group A with acquired B Anti A, B cells, Anti B

Immunogenecity D>c>E>C>e
Rh Ag are proteins
Kleihauer Betke Test Fetomaternal Hemorrhage
Flow Cytometry
Fetal RBCs (high Hb F) Resists acid elution – DEEP PINK
Maternal RBCs Eluted from the cell – GHOST CELLS
ABO HDN Rh HDN
Most common Severe
Cannot be diagnosed Immune exposure (second child)
Affect 1st child Very strong DAT (sensitizing Ab)
Weak-to-negative DAT Exchange transfusion
(ABO hemolysis) bind complement
Treatment: phototherapy
KELL (K) is second to D in immunogenicity k- cellano (high frequency Ag present 99.9)
 K – Kell rare (absent in 90% of population, 10% K +)

Lewis Aantigen are not RBC, but produced by Adsorbed from plasma onto rbc membrane
tissue cells
First: Le A
Anti I Enhance: incubation 4C or enzymes-treated RBCs
may be required to detect reactivity
Anti i IM and alcoholic cirrhosis (EBV)
Duffy Resistance to malaria
Fy (a-b-) Resistant to invasion by P. vivax merozoites and P.
Duffy a and b knowlesi
M-N- cells En (a-), resistant to invasion by P. falciparum
merozoites
Anti-M Anti-M: Acidification of px serum
pH of 6.5
Jk (a-b-) RBC resistant to tumor urea
JK (a-b-) 2M urea
Lutheran Blood group Lupus Erythematosus
Anti Lu A (Luteran)
Diego Ag Mongolian ancestry, Asian people and native
south American Indians
XG Antigen Female (89%) > male (66%)
Short arm of X chromosome
Destroyed/inactivated by enzymes
Sex-linked
SC2 (Scianna 2) Mennonite population
Chido Rogers 9 AG attached to the complement
Gerbich or Leach Weak expression of Kell blood group Antigens
Ge 2-3-4
Elliptocyte
Knops system Ag CR1 complement receptor
Sd (a+) Mixed fieald, refractile agglutinate
Urine pang neutralize
030 Rh- associated Glycoproteins( newest blood
group )
Antibodies that exhibit Dosage Rh
Kidd
Duffy
MNS
Lutheran
Dosage Stronger reaction with homozygous
Anti M M+ , N- strongest

Ehannced Rh kidd lewis P1, I ABO

Inactivated Xg, Duffy, MNS

Allogeneic whole blood donations AABB: 8 weeks
 DOH: 12 weeks/3 months
Double rbc pheresis 16 weeks
Apheresis plt, plasma, wbc 2 days
Teratogenic Drug :

Isotretinoin/ Accutane 1month

Proscar, Propecia 1 month
Avodart 6 months
Soriatane 3 years
Tegison Permanent

Hepa B immune globulin (HBIg) 12 months

Px exposed to Hepa B
Experimental medication or unlicensed 1 year
experimental vaccine
Termination of pregnancy 6 weeks
Transfusion during pregnancy 12 months
Abortion or miscarriage (first trimested or second No deferral
trimester)
Creutzfeld-Jakob and Pituitary growth hormone of Permanent
human origin
Recombinant growth hormone No deferral
Vaccines
Measles (measles rubeola, mumps, polio oral, 2 weeks
typhoid, yellow fever
Rubella, german measles, chickenpox 4 week
Polio injection No deferral
Vaccine: Hepa B, A, No deferral
Smallpox
Male to male sex 12 months
Community sex worker / prostitute Permanent
Sexual contact with prostitute for sex 12 months
Treated for syphilis or gonorrhoea 12 months after completion of therapy
Drug IV abuser Permanent
Parenteral drug abuse (needle marks)
History of haemophilia A or B, VWD, or severe Permanent
thrombocytopenia
Viral hepatitis after 11th birthday Permanent
Positive Hepa B (HBsAg) Permanent
Close contact Household or sexual contact with 12 months
indiv with heap B or Hepa C
Reactice anti HBc on >1 occasion Permanent indefinite
Donor bluish or purplish spots on skin (Kaposi Permanent
sarcoma) HIV
Sex w HIV 12 months
Close contact with indiv with HIV No deferral
History of babesiosis or Chagas’ disease Permanent
History of leishmaniasis 12 months
Born or lived in Africa on FDA list of countries with Indefinite
HIV 1 group O risk since 1977
Cancer,leukemia, lymphoma Indefinite deferral
DOH
Dengue 1 mos
Dengue with FFP 12 months
DM: Type 1 Permanent
DM type 2 No deferral
Dental surgery 12 months
Tooth extraction 3 days (72 hours)
Tuberculosis 2 years after complete recovery
Mumps, measles, rubella (MMR) vaccination 8 weeks

Mumps measles rubeola (2 weeks)

Rubella (4 weeks)
MMR (2 + 2+ 4 = 8 weeks)
Malaria 3 years
Immigrants citizens reside in endemic malaria for 5 3 years
years
Traveler of an endemic place of malaria 1 year
Stay less than 6 mos 6 mos
COVID history travel 28 days
Vaccine COVID
Plateletpheresis Aspirin: 3 days/72 hrs
Or 2d 48 hrs
Feldene 2 days
Clopidogrel (plt) 14 days (DOH: 7 days)
Ticlid 14 days
Heparin Prevent coag
Shift to left
Heavy spin 5000g 5 mins
Whole blood ACD, CPD, CP2D: 21 DAYS
Storage: 1-6 C CPDA-1: 35 DAYS
Transport: 1-10
RBC ACD, CPD, CP2D: 21 DAYS
Storage: 1-6 C CPDA-1: 35 DAYS
Transport: 1-10 AS-1, AS-3, AS-5: 42 DAYS

Additive solution: after removing plasma

CPDA-1 200-250 mL plasma can be removed
Frozen rbc -65 C or 120 C: 10 years from collection
High glycerol Frozen at -80 C
Stored at -65 C
Low Glycerol (Liquid N2) Frozen at -196 C
 Stored at -120C
RBCS, DEGLYCEROLIZED OR WASHED (OPEN 1-6 C: 24 hours
SYSTEM )

After removal glycerol (80%) of original RBC mass

Irradiated RBCs : inactivates T cell 1-6 C
Radiation source: cesium/cobalt
28 days from irradiation or original outdate,
whoever is first
Leukopoor RBC 1-6 C
Centrifugation, filtration, saline washing Open system: 24 hours
Closed system: same as WB
Leukoreduced red cells Reduced to 5x10 6 at least 85% of the original RBC
mass
FFP
-18 C: 1 year
-65 C : 7 years
FFP thawed at 37C 1-6 C storage , used within 24 hours
Cryoprecipitate AHF -18 C or colder, 1 year
Thawed Cryo (37C) 20-24C, 6 hours
Pooled cryo 20-224 C, used within 4hours
Components of FFP Components of cryoprecipitated AHF
- Plasma, all coagulation factors, - Fibrinogen (min. 150 mg , QC 250 mg)
complement - AHF (VIII) 80 U
- vWF
- Factor XIII
- Fibronectin
Pooled platelet 20-24 C : 4 hours
Platelet 20-24 C 5 days
Pooled platelet
Apheresis granulocyte (Leukapheresis CSF) 20-24 C: 24 hours
Hydroxy
Sedimentation agent
1 SDP (A pheresis donor platelets) 4-6 random donor platelet (RDPs)
Plateletpheresis 500 mL plasma removed

600mL for donors more than 175 lbs weight

Prepare packed rbc: 200-250 mL plasma removed
Platelet concentrate Red cell ( more than 2 ml)  crossmatch
RHOGAM Component: IgG anti D
Prevention of rH HDN
Passive immunity (antibody
Administered within 72 hours after delivery ( 3
days)
Storage: 1-6 C
Shelf life: 2 years
Prevention of rh HDN
 Treatment of Immune thrombocytopenic purpura
Neocyte Young rbc (reticulocytes)
Separated using density
Mature liit but heavy
Immature big but gaan
Longer lifespan to reduce frequency of transfusion
Transfusion dependent patients
1. Aplastic anemia
2. Congenital haemolytic anemia
Aplastic anemia Blood: Leukoreduced rBC to decrease HLA
exposure
Washed rbc less than 7 days
PNH Washed rbc or frozen, deglycerolized rbc
Double rbc pheresis 2 weeks of rbc are collected by apheresis, donor
2RBC must wait 16 weeks before providing another
donation that includes rbcs
Male:
- >130lbs
- 5’1 height
- 40% hct
Female
- >150 lbs
- > 5’5
- %40 hct
16 weeks deferral
Nurses: final clerical check
Fatal Transfusion reactions Mostly caused by Clerical Errors
Most serious haemolytic transfusion ABO
KIDD Delayed haemolytic transfusion reactions
Most common cause of fatal transfusion rxns Clerical error
Most common transfusion rxns: FNHTR and allergic rxns
Most common type of trans rxns associated with TACO and TRALI
mortality (death)
Most Severe HDN Anti-D
D is the most antigenic
ABO HDFN is more common than RH HDN and
produce no symptoms
Febrile nonhemolytic reaction Increase in temp 1C or more
Anti leukocyte Antibodies
Transfuse Leukocyte reduced products (2 or more
febrile nonhemolytic transfusion rxns)
Leukopoor <5 x 10 6
Recover 85% RBC
TRALI Transfusion related acute lung injury
Leukocyte Antibodies (donor)
Parous female donors
 Plasma from male donors
Antileukocyte antibodies
Platelets stored at room temp – prone to bac Bacterial contam
contam
Yersinia enterocolitica – most common isolate
found in rbc units (ref) followed by Pseudomonas
putida / fluorescence
Propionibacterium acnes – skin (subacute bacterial
endocarditis) most common bacterial contaminant
in rbc
Staph epidermidis and B. cereus (G+) are
organisms most frequently recovered from
donated blood
Endocarditis Catalase and spot indole test : P. acnes is positive
T. pallidum Dies in refrigeration of stored blood for 3 days
Platelet concentrate Responsible for transmitting syphilis
Syphilis RITM : for confirmatory of syphilis
Transfusion-transmitted parasites Babesia microti
Trypanosome cruzi
Malaria (plasmodium species) – most common
parasite in blood transfusion
Babesia microti Survive in refrigerated uncoagulated blood for 21
days to 35 days
Preferred stain: Giemsa
TACO Iatrogenic type of transfusion reaction (physician
Transfusion associated Circulatory Overload cause)

Test: BNP (Brain natriuretic peptide)

NT-proBNP
BNP 1.5 times higher: diagnostic of TACO
Graft vs host disease Delayed immune transfusion reaction
Irradiated
T cell
Cobalt or cesium
SOP (standard operating procedure) Document
Reviewed annually
Worksheet Record
Pt record
3 major component of computer Hardware, software, peopleware

IMMUNOLOGY AND SEROLOGY

Skin ph prone to bacterial contamination pH: 5.6
Moist skin
Interferon (IFN) Antiviral agents
Immunomodulators
Antineoplastic agents (anti cancer)
IFN-a Treatment hepa C, Kaposi’s sarcoma, leukemias ,
 and lymphomas
IFN B Treatment multiple sclerosis
Marginal B cell Spleen
Follicular B cell Secondary lymphoid organs
KIR (Killer immunoglobulin-Like receptor system NK lymphocyte – mediate graft-vs-leukemia (GVL)
reaction and prevent relapse after transplantation
for hema malignancies
DAT Detect in vivo sensitization of rbc
Invest HTR
Diagnosis HDN, AIHA, drug-induced hemolysis

PINK STOPPER EDTA

IAT In vitro
Crossmatchh=
Red cell antigen typing
Ab detection/ab screen
Ab ID

Pat serum of rbc

RED STOPPER
Dithiothreitol and 2-mercaptoethanol Break disulphide bonds of J (joining) chain of IgM
to detect IgG antibodies
Monoclonal antibodies Milstein and Kohler
Very specific reagent of in vitro diagnosis
Current use: THERAPEUTIC AGENT
- Rheumatoid arthritis
- Crohn’s disease
- Anti-cancer
Mouse spleen  myeloma cells in presence of
PEG (polyethylene glycol)  PEG and plasma cell =
HYBRIDOMA
HYBRIDOMA Immortal cells that produce monoclonal
Antibodies
HLA PHENOTYPING Complement-dependent cytotoxicity
Anticoagulant: ACD (yellow stopper)
Inverted phase-contrast microscope
Method of choice: Molecular studies / tests
Autoimmute diseases Systemic
- SLE (dsDNA)
- Rheumatoid Antibodies ( anti CCP) cyclic
citrulinated protein
- WEGENERS GRANULOMATOSIS (anca ab)
antineutrophilic cytoplasmic antibodies
Systemic : SLE, RA, WEGENERS GRANULOMATOSIS Organ specific : hashimoto’s disease
ANA TEST Hep2 cells human epithelial cells
FANA Mouse kidney
Specific test for anti-dsDNA Substrate: C. luciliae
Primary Syphilis IgM: first ab
Secondary syphilis IgG
Neurosyphilis CSF: VDRL Venereal disease research laboratories
Most likely to be positive in tertiary syphilis FTA-ABS

VDRL and RPR and TRUST: NonTreponemal test

reagin
FTA-ABS Detects: anti-treponemal antibodies
Antigen: Nichol’s strain of Treponema pallidum
Sorbent: Reiter’s strain – removes cross reactivity
FITC-labeled reagent: AHG
Microscope: Fluorescence microscope

Dark-field fluorescent Microscope assembly

CD4 T cell Range: 500 to 1300 cells/uL
AIDS: less than 200 cells/uL
Stage 1 – primary HIV 500 cells/uL
Stage 2 – intermediate HIV 200 to 499 cells/uL
Increase or normal serum globulins
Stage 3 – AIDS <200 cells/uL

Primary Antibodies use to detect H. pylori (Peptic IgG

ulcer) Diagnosed on chronic stage na
Gold Standard for Rickettsial Antibodies Indirect Fluorescent assays (IFA)
Microimmunofluorescent assays (Micro-IF)
IFA Similar to Elisa
Target cells for rickettsial Endothelial cells
Mannan and anti-mannan antibodies Candida species
VZV (Varicella Zoster Virus) Most common : ELISA
REFERENCE METHOD: FAMA (Fluorescent
Antibody to membrane Antigen
Congenital Toxoplasmosis EIA IFA testing widely used, EIA is method of choice
PCR – method of choice to detect T. gondii DNA in
CSF
Autofluorescnece Cyclospora cayetanensis
Fruits: raspberries
Target Amplification PCR
Signal Amplification Branched chain signal
Probe amplification Ligase chain reaction

FIX FACTOR IX
Storage of FIX: 1-6 C
ISG Immune serum globulin
Immune or antibody deficient