Bloodbanking Numericals

Common cause of anamnestic response

Kidd

2 laws of Mendel

1. Law of Segregation- two alleles separate during gamete formation

2. Law of Independent Assortment- genes for traits are inherited independent of the others

2023 Nobel Prize for discovering nucleoside modifications to determine mRNA vaccines against COVID-19

Katalin Kariko

Drew Weissman

First blood transfusion

Pope Innocent VII

1492

Edward Jenner

Small Pox Vaccination

1798

Landsteiner discovered ABO

Landsteiner book (SSR)

1901

Specificity of Serologic Reactions

Von Decastello and Sturli discovered AB

1902 ABO

Braxton Hicks

Sodium phosphate

1869

Rous and Turner

citrate-dextrose

1916

Lewisohn

minimum amount of citrate

1915

Hustine

Use of sodium citrate

1914

Incubation time for 22% Bovine Albumin

30-60 minutes

Longest

Dr. Charles Drew

American Red Cross director

1941

Loutit and Mollison

Acid citrate dextrose

1943

Coombs, Mourant, Race discovered AHG reagent in

1945

Gibson

citrate phosphate dextrose

1957

POTENTIATORS

a. Protein Medium

22% Bovine Albumin

PEG

Protamine

Polyvinylpyrrolidone

Albumin incubation time

PEG incubation time

LISS incubation time

30-60 minutes

10-30 minutes

10-15 minutes

POTENTIATORS

c. Use of Enzymes

Enhances:

Destroys:

Enhances: Rh, I, P1, Kidd, Lewis

Destroys: Duffy, MNSs, Xg (+ Chido/Rodgers, Yt)

Washing for Wharton's Jelly

6-8

POTENTIATORS

b. LISS

Concentration:

Incubation time:

Concentration: 0.2% NaCl (hypotonic so add glycine to prevent hemolysis)

Incubation time: 5-15 minutes

POTENTIATORS

d. AHG

Factors affecting AHG test

Cell-serum ratio:

Temperature:

Centrifugation:

Cell-serum ratio: 1: 40 ; 1 drop RBC: 2 drops serum

Temperature: 37 degrees Celsius

Centrifugation: 1000 RCF for 20 seconds

Gel technology

Gel used:

Founder:

Principle:

Advantage:

Disadvantage:

Incubation:

Centrifugation

Stability of reaction:

Gel used: Dextran-acylamide

Founder: Yves Lapierre

Principle: Hemagglutination

Advantage: Standardization

Disadvantage: Expensive centrifuge, incubator, pipet

Incubation: 15 minutes

Centrifugation: 10 minutes

Stability of reaction: 3 days

Gel Technology grading

4+ solid band at top

3+ agglutinates begin to disperse but are concentrated at the top

2+ agglutinates are observed throughout the microtube

1+ concentrated at the bottom agglutinates

Negative: Cell button at bottom

MF: top and bottom

Minimum washes for Anti-globulin tests

3

To remove free globulins

Hemolysis is only seen in

serum, not plasma (EDTA chelates calcium and prevent complement activation)

Single most important reaction in BB

Hemagglutination

Normal serofuge RCF

1000

ABO centrifuge speed and time

1000 RCF for 20 seconds

Serologic Grading

4+ - One solid

3+ - Large agg

2+ - Medium agg

1+ Small agg / turbid

w+ - tiny / turbid

0 - no agg / hemolysis

mf - small agglutinates within predominantyle unagglutinated cells

H - hemolysis

PH - partial hemolysis (intact red cells still in the bottom)

Detection levels of AHG tests

DAT:

IAT:

DAT: 100-500 IgG and 400-1100 C3d

IAT: 100-200 IgG and C3d

Best factor on rouleaux

fibrinogen

Substance that differentiate agglutination and rouleaux

NSS

agglutination - unchanged

rouleaux - dispersed

Chromosome Locations of blood group genes

1 - Rh, Duffy

4 - MNSs

6 - Chido/ Rodgers

7 - Kell

9 - ABO

18 - Kidd

19 - Landsteiner, Lewis, Lutheran, H, Secretors

22 - P1PK

X - Xg

Discrepancy grading

1+ and w+

ISBT number designation

Ang Mens Po ni Rhea Lumabas Kaso Lang Di Kita Diego

001 - ABO

002 - MNS

003 - P1PK

004 - Rh

005 - Lutheran

006 - Kell

007 - Lewis

008 - Duffy

009 - Kidd

010 - Diego

Although IgM, ABO HDFN is not the most common due to the fact that ABO is only fully-developed by

But now it is the most common due to Rhogam uses

2-4 years of age

ISBT has 6 numbers

The first 3 is ______

The last 3 is _____

The first 3 is BLOOD GROUP SYSTEM

The last 3 is ANTIGEN SPECIFICITY

Order of ABO abundancy

O - A - B - AB

Main advantage of ISBT

Machine and eye readable

ISBT ABO

001001 - A

001002 - B

001003 - AB

001004 - A1

ABO incompatibility type of rejection

hyperacute

ABO nomenclature

Landsteiner Jansky Moss

Landsteiner Jansky Moss

O I IV

A II II

B III III

AB IV I

Percentage of ABO in population

O - 45

A - 40

B - 10

AB - 5

Uses of Lectins

Differ A1 and A2

Differ True B and Acquired B

Differ O and Bombay

Dolichus biflorus agglutinate

A1

# of antigens for ABO blood groups

A - 810,000-1,170,000

B - 610,000-830,000

AB - (A)600,000 (B)720,000

O has Anti-A,B which is

IgG

Reagent for reverse typing

4-5% red cell suspension

tomato red

Order of adding reagents

Antisera / serum (clear first) then RC suspension/ reagent RBC (turbid second)

Reverse typing is not required for:

< 4 months infant

previously typed donor cells

How many phenotypes in ABO

5

A

B

AB

O

Bombay

Acquired A (2)

P. mirabilis infection

Tn-activated erythrocytes

Immunodominant sugars of ABO

A - N-acetyl-D-galactosamine

B - D-galactose

H - L-fucose

Acquired A can happen to ___ persons

Acquired B can happen to ___ persons

B, O

A

Acquired B (4)

EPIC

E. coli O86

P. vulgaris

Intestinal obstruction

Carcinoma of colon and rectum

Main pathology of Acquired B

Deactylase enzyme from bacteria can remove "acetyl of N-acetylgalactosamine (immunodominant sugar of A) to for galactosamine which looks like galactose. So A can become B. O persons cannot have acquired B since they do not have the sugar.

6 detection methods for Acquired B

Diagnosis

Monoclonal anti-B

acidified anti-B

anti-B lectin

Acetic anhydride (acetylates back)

Secretor status

Subgroups are the result of

less effective enzymes. Not efficient in converting H antigens to A or B antigens

A2 subgroup has ___ reactivity to H antigens than A1

higher (A1 converted every H antigen to either A or A1)

Test for differentiating A subgroups

Secretor studies

Adsorption-elution

Molecular studies

A subgroups

Anti-A1 presence or absence

Present - A2, Ax, Aend, Ael

Abundance of antigen sites (x 10 ^ 3)

A3 (35) > Ax (5) > Aend (3.5) > Am (1) > Ay (1) > Ael (0.7)

B subgroup with anti-B

Bx and Bel

H types

H1 and H2 - linear

H3 and H4 - branched

Precursor substances and genes that control it

Type 1 (secretions)

Type 2 (RBCS)

Type 1 - D-galactose is linked with N-acetylglucosamine in B 1-3 linkage

Type 2 - D-galactose is linked with N-acetylglucosamine in B 1-4 linkage

Type 1: ABH, Se, Le

Type 2: ABH

Secretor and nonsecretor percentage

Secretor (80%)

Nonsecretor (20%)

All precursor sugar are ________

All precursor chains have this terminal sugar

Hexoses

Galactose

How to determine the possible genotype and phenotype of offspring using parent's genotype

Mother (AA, AO)

Father (OO)

AA, AO x OO

By multiplying those you get:

6 AO and 2 OO = 8 total genotypes

A phenotypes: 6 AO/ 8 Total = 75% will be of A phenotype

O phenotypes: 2 OO/ 8 Total= 25% will be of O phenotype

Who elucidate the subgroups of A and B

Von Dungern and Hirzfield

H abundancy in order

O > A2 > B >A2B > A1 > > A1B

ABO subgroups differ in the

amount of antigens present on RBCs

A1 and A2 percentage

A1 (80%), Branched, 2 million A antigens

A2 (20%), Linear, 500,000 A antigens

H deficient phenotype first reported by Bhende in

1952

A subgroup that produces anti-H

A1

Group 1 discrepancy (most common)

reverse typing

Cause:

decreased antibodies

dilutional

Rare Group 1 discrepancy

Chimerism - presence of two cell population in an individual

Best division of Weak subgroups of A

Agglutinated by Anti-A or Anti-AB:

A3: mixed field for both

Aend: mixed field, weak for both

Ax: only with anti-AB

Not agglutinated by Anti-A or Anti-AB:

Am: AH substance

Ay: Small AH substance

Ael: H substance

Best division of Weak subgroups of B

Agglutinated by Anti-B or Anti-AB:

B3: mixed field

Bx: only with anti-AB

Bm: BH substance

Bel: B substance

Most common and least common discrepancy of ABO

Most common: Group 1

Least common: Group 2

Phase of antibody screening with no rouleaux

AHG phase (washing is done)

Group 2 discrepancy (least common)

forward typing

Subgroups of A and B

Leukemia (Hodgkin's)

Acquired B

BGSS

Antibody in Acquired B

pH in Acquired B

Anti-B clone ES4

>8.5 or <6.0

Resolution of Type 1

Extend reverse typing:

RT for 15-30 minutes

4C for 15-30 minutes

Group 3 discrepancy

rouleaux:

Increased globulins (MM, WM, Hodgkin's lymphoma)

High fibrinogen

Plasma expanders (dextran and PVP)

Wharton's Jelly

Resolution of Type 2

Extend reverse typing:

RT for 15-30 minutes

4C for 15-30 minutes

For BGSS: Wash red cells

Group 4 Discrepancy

1,2,3 unclassifiabl due to miscellaneous reasons:

Autoantibodies

Transplant/Transfusion (more than 1 ABO group)

Alloantibodies

ABO isoagglutinins

Rare group 4

CIS-AB

Resolve Type 3

Wash 2-3x (6-8x if Wharton's Jelly)

Main use of Anti-AB (2)

1. Check reactions of A and B

2. Detect Weak Subgroups of A and B since it has high titer of Anti-A and Anti-B

If discrepancy is not resolved, transfuse

O-negative red cells

4 examples of amorph gene

O

Lu

Jk

d

Anti-rhesus ab (Landsteiner and Weiner) agglutinated __% of human RBC

85

Procedure if a patient is anti-AB positive

1. Perform saline control on room temperature

Order of Immunogenicity

D>c>E>C>e

Rh nomenclature

Fisher race versus Weiner (Rh-Hr)

D - Rh0

C - rh'

E - rh''

c - hr'

e - hr''

Modified Weiner nomenclature

example: Ro/r'

R

r

1 or '

2 or ''

no 1 or '

no 2 or ''

Rz

ry

Rh nomenclature

Fisher race versus ISBT

D - 004001

C - 004002

E - 004003

c - 004004

e - 004005

Rh nomenclature

Fisher race versus Rosenfield

D - Rh1

C - Rh2

E - Rh3

c - Rh4

e - Rh5