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MTAP 1 > IMMUNOHEMATOLOGY > Flashcards

IMMUNOHEMATOLOGY Flashcards

1
Q

Gel test was invented in ______ by ________.

A

1985, Dr. Yves Lapierre

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2
Q

Two antigen/antibody reactions

A

agglutination and hemolysis

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3
Q

In gel testing, ________ is detected however hemolytic reaction is not detected

A

hemeagglutination

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4
Q

PARALLEL TESTING

A

gel test + tube test

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5
Q

immortalized antibody producing cell

A

hybridoma cell

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6
Q

the antibody producing capacity of hybridoma cell is due to

A

plasma cell

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7
Q

what is elution

A

detachment/dissociation of antigen or antibody

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8
Q

IMPORTANCE OF WASHING IN DAT

A

(1) presence of other unbound antibodies
(2) washing phase
(3) addition of AHG
(4) IgG bridge/ lattice formation
(5) agglutination

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9
Q

Steps in Direct AHG

A
  1. Washed patient’s red cell (3x), sensitized in vivo
  2. Addition of AHG reagent (Coomb’s sera)
  3. Visual red cell agglutination
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10
Q

Steps in Indirect AHG for human red cells

A
  1. Human red cells + Human (patient serum) IgG antibody
  2. Red cell (in vitro) sensitization
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11
Q

Steps in Indirect AHG for sensitized red cells

A
  1. sensitized red cells + Coomb’s sera AHG reagent
  2. Visual red cell agglutination
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12
Q

Gel test principle

A

size exclusion chromatography

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13
Q

what gel is used in gel test

A

dextran polyacrilamide gel

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14
Q

steps/procedure in gel test

A
  1. addition of cells
  2. addition of plasma/serum
  3. incubation for IgG - body temp
  4. centrifugation - 5 mins
  5. results - more than 10 mins
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15
Q

the bigger the size, the ______ the position of cells in the medium

A

higher

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16
Q

reagent is already pre-dispensed, only add the sample to microtube
a. specific
b. low ionic antiglobulin
c. neutral/plain

A

a

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17
Q

reagent is not added to the gel, it is added along with the sample in microtube
a. specific
b. low ionic antiglobulin
c. neutral/plain

A

c

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18
Q

specific for sensitization test. IgG antibodies are detected. AHG reagent is pre-dispensed in gel matrix
a. specific
b. low ionic antiglobulin
c. neutral/plain

A

b

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19
Q

advantages of using gel testing

A
  1. standardization
  2. stable and well defined agglutination reaction
  3. decreased sample volume <1mL
  4. enhanced specificity and sensitivity, no need for cell washing
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20
Q

ABO forward/direct cell typing is and example of
a. specific
b. low ionic antiglobulin
c. neutral/plain

A

a

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21
Q

INDIRECT/reverse/backward typing is an example of
a. specific
b. low ionic antiglobulin
c. neutral/plain

A

c

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22
Q

In gel testing, stability is up to _______

A

3 days

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23
Q

In conventional tube method, stability is up to ____________

A

an hour

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24
Q

In slide method, stability is up to _______

A

1 min

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25
Q

If RBC are unagglutinated, it will pass thru the gel and will form __________ at the bottom

A

cell button

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26
Q

one solid clump, 100 percent antigen reacted with antibodies
a. 3+
b. 4+
c. 1+
d. +/-

A

b

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27
Q

several large sized clumps, 75 percent antigen reacted with antibodies
a. 3+
b. 4+
c. 1+
d. +/-

A

a

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28
Q

several medium sized clumps, 50 percent antigen reacted with antibodies
a. 3+
b. 4+
c. 2+
d. +/-

A

c

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29
Q

several numerous small sized clumps, <25 percent antigen reacted with antibodies
a. 3+
b. 4+
c. 1+
d. +/-

A

c

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30
Q

weak reaction, very small sized clumps, <25 percent reacted with antibodies
a. 3+
b. 4+
c. 2+
d. +/-

A

d

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31
Q

RBC on top most part of the cell
a. 3+
b. 4+
c. 2+
d. +/-

A

b

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32
Q

RBC on upper part of gel matrix
a. 3+
b. 4+
c. 2+
d. +/-

A

a

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33
Q

RBC is disseminated on gel matrix with most RBC found at the center
a. 3+
b. 4+
c. 2+
d. +/-

A

c

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34
Q

RBC are found on lower part of the gel
a. 3+
b. 1+
c. 2+
d. +/-

A

b

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35
Q

RBC found near at the bottom of the tube
a. 3+
b. 4+
c. 2+
d. +/-

A

d

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36
Q

What antibody cannot agglutinate on its own and is the smallest so it can cross the placenta

A

IgG

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37
Q

A technique for detecting cell-bound immunoglobulin and is used to detect incomplete antibodies (IgG).

A

AHG Coomb’s test

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38
Q

acts as a bridge/link so IgG can form lattice sequence

A

AHG

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39
Q

color of AHG/Coomb’s test

A

GREEN

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40
Q

Complement binding via classical pathway from MOST - LEAST potent
a. IgG subclass 1, 2, 3
b. IgG subclass 2, 1, 3
c. IgG subclass 3, 1, 2
d. IgG subclass 1, 3, 2

A

c

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41
Q

natural, big pentamere
a. IgG
b. IgM

A

b

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42
Q

complete best agglutinating antibody
a. IgG
b. IgM

A

b

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43
Q

cold-reacting, 1-6 deg
a. IgG
b. IgM

A

b

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44
Q

saline reactive
a. IgG
b. IgM

A

b

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45
Q

ABO Antibodies
a. IgG
b. IgM

A

b

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46
Q

complement binding and is more potent
a. IgG
b. IgM

A

b

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47
Q

Immune
a. IgG
b. IgM

A

a

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48
Q

incomplete, coating/sensitizing
a. IgG
b. IgM

A

a

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49
Q

warm-reacting, 37 deg and is clinically significant
a. IgG
b. IgM

A

a

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50
Q

albumin/AHG reagent
a. IgG
b. IgM

A

a

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51
Q

Rh antibodies
a. IgG
b. IgM

A

a

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52
Q

consists of a pool of rabbit anti-human IgG and mouse monoclonal anti C3b and anti C3d
a. monospecific AHG reagent
b. polyspecific AHG reagent

A

b

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53
Q

Also referred to as broad spectrum coombs reagent
a. monospecific AHG reagent
b. polyspecific AHG reagent

A

b

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54
Q

method of preparation in polyspecific AHG reagents

A

conventional/classic method (hyper immunization of rabbit)

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55
Q

contains only one antibody specificity
a. monospecific AHG reagent
b. polyspecific AHG reagent

A

a

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56
Q

can either be Anti-IgG, Anti-C3b or C3d
a. monospecific AHG reagent
b. polyspecific AHG reagent

A

a

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57
Q

METHOD of preparation in monospecific AHG reagents

A

hybridoma method (KOHLER and MILSTEIN technique)

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58
Q

hybridoma cell is made up of

A

mouse plasma cell AND myeloma cell

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59
Q

first stage in Ag-Ab reaction

A

sensitization

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60
Q

it occurs when antibodies react with antigens on the cells and coat the cells

A

sensitization

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61
Q

antigen binding fragment

A

Fab

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62
Q

determinant, reacts with antibody

A

epitope

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63
Q

second stage in Ag-Ab interaction

A

agglutination

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64
Q

it occurs when abs on coated cells form cross-linkages between cells resulting in visible clumping

A

agglutination

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65
Q

cross linking of Abs adjacent to antigen

A

lattice formation

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66
Q

single most important serologic test to diagnose HDFN

A

DAT

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67
Q

detects in vivo sensitization of red cells with IgG and/or complement

A

DAT

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68
Q

Investigation of transfusion reactions
a. IAT
b. DAT

A

b

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69
Q

diagnosis of autoimmune and drug-induced hemolytic anemias
a. IAT
b. DAT

A

b

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70
Q

cells used for DAT should be collected into either ———— to minimize the possibility of ——- attachment of complement components

A

EDTA or citrate containing anticoagulant
IN VITRO

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71
Q

the ———— is needed to demonstrate antibodies in the event of in vivo erythrocyte sensitization

A

DAT

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72
Q

a two-step procedure that determines in vitro sensitization of red cells
a. IAT
b. DAT

A

a

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73
Q

in IAT, what are the two procedures involved

A

sensitization (incubation/thermophase)
agglutination (AHG step)

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74
Q

incubation/thermophase
a. agglutination
b. sensitization

A

b

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75
Q

AHG step
a. agglutination
b. sensitization

A

a

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76
Q

detection of incomplete antibodies in compatibility testing or to screening cells in antibody
a. DAT
b. IAT

A

b

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77
Q

identification of antigen specificity, using a panel of red cells
a. DAT
b. IAT

A

b

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78
Q

determination of red cell phenotype using known antisera (Du testing)
a. DAT
b. IAT

A

b

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79
Q

titration of incomplete antibodies
a. DAT
b. IAT

A

b

80
Q

used for confirmatory testing for weak D testing

A

IAT

81
Q

In IAT, we use this as confirmatory testing for weak D specifically what type?

A

low grade type

82
Q

report for low grade weak D is

A

Rh positive

83
Q

If anti-D is negative, report immediately
TRUE or FALSE

A

false

84
Q

If IAT is positive it is considered as —-

A

Rh pos because IAT is used to confirm for the presence of low grade Du antigen

85
Q

AHG is also referred to as

A

anti-antibody

86
Q

Minimum ratio of serum to cells

A

40:1, 2 drops serum and 1 drop of 5 percent v/v cell suspension

87
Q

optimal temperature for AHG testing

A

37 degC

88
Q

incubation time for AHG testing in saline suspension

A

30-120 mins

89
Q

incubation time for AHG testing in LISS suspension

A

10-15 mins

90
Q

PROZONE
a. excessive Ag
b. excessive Ab

A

b

91
Q

POSTZONE
a. excessive Ab
b. excessive Ag

A

b

92
Q

in AHG reaction medium, ___ minute saline test = ___ minute albumin test
a. 30, 20
b. 15, 30
c. 30, 60
d. 60, 30

A

d

93
Q

____ is said to reduce the zeta potential between RBCs thus increasing the rate of Ab uptake on the cell

A

22 % albumin

94
Q

____ also increases the sensitivity and shortens incubation times

A

LISS

95
Q

LISS is an example of _____ which enhances ag/ab interaction

A

potentiator

96
Q

in AHG test, saline for washing should be fresh and buffered to a pH of ______

A

7.2-7.4

97
Q

Addition of AHG reagents should be added to washed cells immediately after washing
TRUE or FALSE

A

True

98
Q

In AHG test, centrifugation is done for

A

1000 rcf for 15-20 seconds

99
Q

factors affecting AHG test

A
  1. ratio of serum to cells
  2. temp
  3. incubation time
  4. reaction medium
  5. washing of cells
  6. saline for washing
  7. immediate addition of AHG reagent
  8. centrifugation
100
Q

Incomplete washing
a. neg
b. pos

A

a

101
Q

extreme pH (cause protein denaturation)
a. neg
b. pos

A

a

102
Q

too fast, too long
a. false pos
b. false neg

A

a

103
Q

too slow, too short
a. false pos
b. false neg

A

b

104
Q

spontaneous elution
a. false pos
b. false neg

A

b

105
Q

autoagglutinable cells
a. false pos
b. false neg

A

a

106
Q

bacterial contamination or other contamination in cells or saline
a. false pos
b. false neg

A

a

107
Q

cells with a (+) DAT used for IAT
a. false pos
b. false neg

A

a

108
Q

overcentrifugation or overreading
a. false pos
b. false neg

A

a

109
Q

polyagglutinable cells
a. false pos
b. false neg

A

a

110
Q

dirty glasswares
a. false pos
b. false neg

A

a

111
Q

saline contaminated with silica or heavy metals
a. false pos
b. false neg

A

a

112
Q

specificity is DECREASED
a. false pos
b. false neg

A

A

113
Q

sensitivity is DECREASED
a. false pos
b. false neg

A

b

114
Q

inadequate or improper washing of cells
a. false pos
b. false neg

A

b

115
Q

most common cause of false neg in AHG technique

A

inadequate or improper washing of cells

116
Q

AHG reagent nonreactive owing to deterioration or neutralization
a. false pos
b. false neg

A

b

117
Q

AHG reagent not added
a. false pos
b. false neg

A

b

118
Q

serum not added in the indirect test
a. false pos
b. false neg

A

b

119
Q

serum non reactive owing to deterioration of complement
a. false pos
b. false neg

A

b

120
Q

inadequate incubation conditions
a. false pos
b. false neg

A

b

121
Q

postzone and prozone (cell suspension either twoo weak or too heavy)
a. false pos
b. false neg

A

b

122
Q

undercentrifugation
a. false pos
b. false neg

A

b

123
Q

poor reading technique
a. false pos
b. false neg

A

b

124
Q

to allow sensitization reaction, contains agent to promote interaction of red blood cell
a. LIP- low ionic polybrene technique
b. ELAT- enzyme linked antiglobulin test
c. Solid phase method - direct/indirect

A

a

125
Q

similar to ELISA, uses RBC (contains IgG). Its primary antibody uses AHG.
a. LIP- low ionic polybrene technique
b. ELAT- enzyme linked antiglobulin test
c. Solid phase method - direct/indirect

A

b

126
Q

occurs in microwells, Antigen are bound to bottom of well and patient’s plasma/serum is incubated in the well
a. LIP- low ionic polybrene technique
b. ELAT- enzyme linked antiglobulin test
c. Solid phase method - direct/indirect

A

c

127
Q

substrate changes in color when there is AHG sensitized red cell binding.
a. LIP- low ionic polybrene technique
b. ELAT- enzyme linked antiglobulin test
c. Solid phase method - direct/indirect

A

b

128
Q

in ELAT, color of substrate is _________ to quantity of sensitized Antibodies present in sample

A

directly proportional

129
Q

uses colorimeter as principle
a. LIP- low ionic polybrene technique
b. ELAT- enzyme linked antiglobulin test
c. Solid phase method - direct/indirect

A

b

130
Q

used to detect as many clinically significant antibodies outside ABO system

A

antibody screening procedure

131
Q

in antibody screening procedure, ________ is used to differentiate different types of antibodies

A

enzyme procedure

132
Q

enzyme reagents

A

FICIN, BROMELIN, PAPAIN, TRYPSIN

133
Q

for IgG alloabs (qualitative)

A

patient serum + screening cells

134
Q

for autoantibodies

A

patient serum + red cells

135
Q

______________ now replaces the minor cross-matching procedure in routine pre-transfusion

A

antibody screening procedure

136
Q

what blood type is used to as screening cell to prevent ABO abs from producing reactions

A

O

137
Q

if the antibody screen is reactive, this must be determined

A

antibody specificity

138
Q

panel used for antibody identification

A

11 reagent panel

139
Q

use of ____, _____ and ______ will inactivate some antigens specially ________.

A

AET, DTT and ZZAP
KELL

140
Q

in the prewarm procedure, this may be removed

A

clinically insignificant cold antibodies

141
Q

in the prewarm procedure, patient serum, reagent red cells and enhancement medium can be warmed separately at

A

37 degC for 5-10 mins prior to mixing

142
Q

these reagents denatures IgM antibodies by breaking disulfide bonds

A

sulfhydryl or thiol reagents *DTT and 2-ME

143
Q

use of _____________ techniques to remove unwanted antibodies such as cold or warm autoantibodies, or to help resolve multiple antibodies

A

adsorption and elution

144
Q

used to remove unwanted antibodies from Serum
a. elution
b. adsoprtion

A

b

145
Q

used to dissociate IgG abs from sensitized cells
a. elution
b. adsoprtion

A

a

146
Q

this may also be used as adsorbents for anti-I since they are rich in I antigen.

A

rabbit cells

147
Q

these autoantibodies can be adsorbed onto the patient’s enzyme pretreated cells at 4 degC

A

I, H, or IH

148
Q

the recovered antibody, eluate can be tested like ____ to determine antibody’s _____

A

serum, specificity

149
Q

contains detached antibodies

A

eluate

150
Q

example of elution techniques

A

heat, freeze-thaw, use of organic solvent, acid eluates, using ZZAP or chloroquine diphosphate

151
Q

mixture of DTT and papain that is used to remove Ab from sensitized red cells and to enzyme treat them at the same time
a. AET
b. DTT
c. ZZAP
d. chloroquine disphosphate

A

c

152
Q

reagent used to remove IgG Abs from the surface of sensitized cells; inactivates Bg antigens
a. AET
b. DTT
c. ZZAP
d. chloroquine disphosphate

A

d

153
Q

landsteiner-miller heat
a. first generation
b. second generation
c. third generation

A

a

154
Q

lui-freeze thaw
a. first generation
b. second generation
c. third generation

A

a

155
Q

uses organic solvents
a. first generation
b. second generation
c. third generation

A

b

156
Q

uses non-hazardous chemical agents: acid elution
a. first generation
b. second generation
c. third generation

A

c

157
Q

another technique for facilitating antibody identification is

A

NEUTRALIZATION

158
Q

Hydatid cyst fluid
a. anti P-1
b. anti I
c. anti ABH
d. anti Lea and anti Leb

A

a

159
Q

Plasma or serum with Le subs
a. anti P-1
b. anti I
c. anti ABH
d. anti Lea and anti Leb

A

d

160
Q

Pooled serum or plasma
a. anti P-1
b. anti I
c. anti chido anti rogers
d. anti sda, tamms horsefall protein

A

c

161
Q

urine
a. anti P-1
b. anti I
c. anti chido anti rogers
d. anti sda, tamms horsefall protein

A

d

162
Q

saliva of ‘secretors’
a. anti P-1
b. anti ABH
c. anti chido anti rogers
d. anti sda, tamms horsefall protein

A

b

163
Q

human milk
a. anti P-1
b. anti ABH
c. anti I
d. anti sda, tamms horsefall protein

A

c

164
Q

in heat elution, it will produce

A

dissociated Abs and destroyed erythrocytes

165
Q

in ether elution, name the parts in the tube

A

upper part- ether
middle- destroyed erythrocytes
lower part- dissociated abs

166
Q

pre-transfusion procedure, composed of series of tests to ensure the safety of the recipient during blood transfusion, to select the appropriate blood unit for transfusion

A

compatibility/crossmatch

167
Q

in compatibility/crossmatch, what is the preferred specimen

A

serum

168
Q

in compatibility/crossmatch, why is serum preferred than plasma?

A
  1. may cause small fibrin clots, difficult to distinguish from true agglutination
  2. may inactivate complement, antibodies will not be detected
169
Q

in compatibility/crossmatch, age of specimen must be

A

as fresh as possible

170
Q

in compatibility/crossmatch, age of specimen if the patient has been transfused or pregnant within the past 3 months must be

A

less than 3 days old

171
Q

in compatibility/crossmatch, the sample storage required by AABB must be stored between

A

1-6 degC for at least 7 days after transfusion

171
Q

in compatibility/crossmatch, the sample storage required by AABB must be stored between

A

1-6 degC for at least 7 days after transfusion

172
Q

most critical pretransfusion serologic test

A

ABO grouping

173
Q

if the patient’s ABO group cannot be satisfactorily determined and immediate transfusion is essential, what blood type should be utilized

A

blood type O

174
Q

if Rh type of the recipient cannot be determined and transfusion is essential, what Rh blood type should be given

A

O -

175
Q

routinely done on crossmatching
major or minor

A

major

176
Q

Major x-match
a. Donor’s serum + recipient’s cells
b. Donor’s cells + recipient’s serum

A

b

177
Q

Minor x-match
a. Donor’s serum + recipient’s cells
b. Donor’s cells + recipient’s serum

A

a

178
Q

Patient’s serum + donor’s red cells are tested in saline medium. IgM antibodies are detected
a. AHG technique
b. Broadspectrum technique
c. Saline technique
d. High protein/albumin technique

A

c

179
Q

PS+DR tested in high protein media. IgG abs are detected (Rh Abs)
a. AHG technique
b. Broadspectrum technique
c. Saline technique
d. High protein/albumin technique

A

d

180
Q

PS+DR tested in AHG medium. Non-agglutinating IgG are detected
a. AHG technique
b. Broadspectrum technique
c. Saline technique
d. High protein/albumin technique

A

a

181
Q

composed of 3 stages; (1) IS (2) Thermophase/Incubation (3) AHG
a. AHG technique
b. Broadspectrum technique
c. Saline technique
d. High protein/albumin technique

A

b

182
Q

immediate spin in saline detects

A

IgM

183
Q

thermophase/37degC incubation detects

A

IgG

184
Q

Check cells/Coombs control cells (IgG sensitized cells) should be added to tubes that demonstrate no agglutination. For results to be considered valid, _____ must occur.

A

agglutination

185
Q

invalid check cell is due to

A

failure to wash cell

186
Q

unbound Abs neutralized AHG is due to

A

reagent already deteriorating, not reacting

187
Q

What is the possible problem in this crossmatch
Ab screen: (-)
AC: (-)
Major Xmatch: (+)

A

ABO/Rh typing error
Donor unit with (+) DAT
Patient with low incidence Ab

188
Q

What is the possible problem in this crossmatch
Ab screen: (+)
AC: (-)
Major Xmatch: (+)

A

patient alloantibody

189
Q

What is the possible problem in this crossmatch
Ab screen: (+)
AC: (+)
Major Xmatch: (+)

A

patient autoantibody
rouleaux

190
Q

substances that are able to carry oxygen in the absence of intact red cells

A

blood substitutes

191
Q

common antibody encountered in BB

A

anti big K

192
Q

______ is tested when AHG is negative

A

check cell

193
Q

advantages of SFHS

A

longer shelf life
very stable
not immunogenic
no req for blood typing procedures

194
Q

disadvantages of SFHS

A

short intravascular halflife
possible toxicity
high O2 affinity
high oncotic effect

195
Q

advantages of PFCs

A

biologic inertness
not immunogenic
easily synthesized

196
Q

disadvantages of PFCs

A

adverse clinical effect
high O2 affinity
retention in tissues

MTAP 1 (10 decks)

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