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Transfusion > Week 5 > Flashcards

Week 5 Flashcards

Antibody Detection

1
Q

What does a basic Type and Screen test include?

A

It includes ABO/Rh testing and screening for other blood group antibodies.

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2
Q

When is a Type and Screen test usually performed?

A

It is usually performed when requested by a physician if a transfusion might be required.

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3
Q

Why is a Type and Screen test done during prenatal testing?

A

To assess ABO/Rh compatibility and screen for any antibodies that could affect the pregnancy.

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4
Q

What is assessed in neonates using a Type and Screen test?

A

Hemolytic disease of the fetus and newborn (HDFN), ABO/Rh grouping, and Direct Antiglobulin Test (DAT) testing.

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5
Q

In what emergency situation might a Type and Screen test be necessary?

A

It is performed during emergencies or trauma when a transfusion is likely.

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6
Q

What is the typical outcome of a Type and Screen test for most patients?

A

Most patients will have a normal ABO/Rh type and no antibodies present in plasma.

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7
Q

When are serological investigations required after a Type and Screen test?

A

If any problems arise with the ABO/Rh typing or antibodies are detected in the initial screen.

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8
Q

What suspected issue might prompt pre- and post-samples in a Type and Screen test?

A

Suspected transfusion reactions.

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9
Q

Who else undergoes Type and Screen tests besides patients?

A

Blood and plasma donors, who are tested at CBS before distributing to hospitals.

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10
Q

What genetic mutation can lead to abnormal antigens affecting ABO/Rh testing?

A

Genetic mutations resulting in Weak D antigen.

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11
Q

How can disease states impact ABO/Rh testing?

A

Disease states can result in changes to antigens, making them abnormal or absent.

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12
Q

What issue can arise with antisera during ABO/Rh testing?

A

Antisera may cross-react with other antigens, affecting test accuracy.

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13
Q

What laboratory testing issues can arise in ABO/Rh testing?

A

Cold agglutination or warm autoantibodies can delay test results.

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14
Q

What technique may be used to address ABO/Rh testing issues in patients?

A

The prewarm technique can be used to identify the least incompatible blood for patients with warm autoantibodies.

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15
Q

How can ABO/Rh testing issues impact patient care?

A

Delays in identifying blood group or antibodies can affect the readiness of blood for the patient.

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16
Q

What can be done in an emergency if ABO/Rh testing is delayed?

A

Uncrossmatched blood can be given in an emergency situation.

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17
Q

What is polyagglutination?

A

It is a condition in which red cells are agglutinated by all human sera.

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18
Q

What causes polyagglutination?

A

It is caused by exposure to a normal “hidden antigen” or a red cell antigen mutation.

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19
Q

How do red cells with polyagglutination react when tested with human antisera?

A

They show agglutination regardless of the antibody specificity.

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20
Q

How are patients with polyagglutination affected in ABO/Rh testing?

A

They may be grouped as A and B Rh-positive even if they do not have those antigens.

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21
Q

What is acquired T activation, and what causes it?

A

Acquired T activation is caused by a bacterial or viral infection, such as Streptococcus pneumoniae in children.

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22
Q

What are alloantibodies?

A

Alloantibodies are antibodies produced as a normal immune response to non-self antigens.

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23
Q

Why is a patient’s history of transfusions and pregnancies important in antibody investigations?

A

A history of transfusions and pregnancies is essential because these experiences can expose the patient to foreign antigens, increasing the likelihood of antibody production.

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24
Q

Why is patient diagnosis important in antibody investigations, particularly for sickle cell patients?

A

Sickle cell patients are more likely to produce antibodies, making patient diagnosis vital information for understanding antibody production risks.

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25
Q

Which antibodies are considered significant in transfusion science?

A

Significant antibodies in transfusion science are those that can cause transfusion reactions or impact patient compatibility, though specific examples depend on individual patient factors.

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26
Q

When are autoantibodies produced?

A

They are produced when the patient’s immune system fails to recognize “self” antigens.

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27
Q

What type of cells do autoantibodies react with?

A

Autoantibodies react with almost all human red cells and are usually non-specific or specific to a high-incidence antigen, like Anti-e.

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28
Q

What are cold autoantibodies, and how do they react?

A

Cold autoantibodies are IgM antibodies that react at room temperature (RT) or 4°C.

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29
Q

What are warm autoantibodies, and at what temperature do they react?

A

Warm autoantibodies are IgG antibodies that react at body temperature.

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30
Q

To which cells do autoantibodies attach?

A

Autoantibodies attach to the patient’s own red cells.

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31
Q

Which test can detect antibodies attached to red cells in vivo?

A

The Direct Antiglobulin Test (DAT) can detect antibodies attached to red cells in vivo.

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32
Q

What are OBG antibodies, and how are they produced?

A

OBG antibodies are red cell immune antibodies and depend on the production and stimulation of IgG or IgM forms.

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33
Q

When are Rh and OBG antibodies expected to be present in a patient’s plasma?

A

Rh and OBG antibodies are generally not expected in a patient’s plasma; if present, they result from immune stimulation due to red cell exposure through pregnancy or transfusion.

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34
Q

What is the first step in the stimulation process of antibody production?

A

Antigens on red cells are introduced to the recipient.

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35
Q

What happens if the recipient lacks the introduced antigen?

A

The immune system recognizes the antigens as foreign.

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36
Q

How does the immune response change upon secondary exposure to the same antigen?

A

Memory cells initiate a faster secondary response, with increased antibody levels.

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37
Q

What is the role of significant antibodies in the stimulation process?

A

Significant antibodies can destroy red cells carrying the foreign antigen.

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38
Q

Which antigens are more immunogenic compared to others?

A

Rh and Kell antigens are more immunogenic than other antigens.

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39
Q

How does the immune system of an individual impact antibody production?

A

The strength and response of an individual’s immune system affect the likelihood and quantity of antibody production.

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40
Q

What is the testing protocol for patients who may be transfused or are pregnant?

A

The protocol is to test all patients who have the potential to be transfused and those who are pregnant.

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41
Q

At what age is reverse ABO testing performed on all patients?

A

Reverse ABO testing is performed on patients over 4 months of age.

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42
Q

When is Weak D testing performed?

A

Weak D testing is performed on Rh-negative babies with Rh-negative mothers and for donor testing at CBS.

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43
Q

Is Rh control routinely done?

A

No, Rh control is not routinely done; it is performed only in cases of discrepancy on D1/D2, AB Rh-positive patients, and during Weak D testing.

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44
Q

What type of testing is performed on donor units in the hospital?

A

Only confirmation testing is performed, with no reverse testing.

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45
Q

Which antibodies are considered non-red cell immune and belong to the Lewis system?

A

Anti-Leᵃ and Anti-Leᵇ

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46
Q

What type of antibodies are Rh antibodies?

A

Rh antibodies are red cell immune.

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47
Q

Which non-red cell immune antibodies are associated with the MNS system?

A

Anti-M and Anti-N.

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48
Q

Are Kell antibodies red cell immune?

A

Yes

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49
Q

Is Anti-P1 non-red cell immune?

A

Yes

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50
Q

Are Kidd antibodies red cell immune?

A

Yes

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51
Q

Which non-red cell immune antibodies are associated with the Ii system?

A

Anti-I and Anti-i.

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52
Q

Are duffy antibodies red cell immune?

A

Yes

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53
Q

Which antibodies are red cell immune and part of the MNS system?

A

Anti-S and Anti-s.

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54
Q

What immunoglobulin types can non-red cell immune antibodies be?

A

Non-red cell immune antibodies can be IgM or IgG.

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55
Q

What immunoglobulin type are red cell immune antibodies typically?

A

Red cell immune antibodies are typically IgG.

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56
Q

Which test uses the patient’s cells with Anti-A and Anti-B reagents and results in agglutination?

A

Forward grouping.

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57
Q

Which test uses the patient’s plasma with A1 and B cells, resulting in agglutination?

A

Reverse grouping.

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58
Q

What is used in the screening and identification of antibodies, involving the patient’s plasma?

A

SCI/SCI panel cells, resulting in sensitization/agglutination.

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59
Q

Who are some manufacturers of screening cells?

A

Screening cells are commercially prepared by manufacturers like Ortho, Bio-Rad, and Immucor (DBL).

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60
Q

Why are Group O cells chosen for screening cells?

A

Group O cells are chosen to prevent a reaction with the patient’s Anti-A, Anti-B, and Anti-A,B (ABO antibodies).

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61
Q

What do the symbols “+” and “0” indicate on an antigram for screening cells?

A

The “+” symbol indicates the presence of an antigen, while “0” indicates the absence of an antigen.

62
Q

What must screening cells include for all significant blood group systems?

A

Screening cells must include positive red cells for all the significant blood group systems.

63
Q

What is the purpose of screening cells in antibody detection?

A

Screening cells must be able to detect all significant antibodies.

64
Q

Why is it preferable for screening cells to be homozygous?

A

Homozygous cells are preferred because they provide the strongest expression of antigens, which helps detect weak antibodies.

65
Q

What initial clues do screening cells provide in antibody investigation?

A

Screening cells provide clues such as dosage, which can begin the antibody investigation.

66
Q

What is the Rh phenotype of Screening Cell 1?

A

Screening Cell 1 is always R1R1.

67
Q

What is the Rh phenotype of Screening Cell 2?

A

Screening Cell 2 is always R2R2.

68
Q

Why is it important to have homozygous cells for each Rh antigen in screening cells?

A

Homozygous cells provide a strong expression for each Rh antigen (C, E, c, and e), aiding in the detection of antibodies.

69
Q

Which Rh antibodies are commonly found in patients?

A

Antibodies to C, E, and c are common.

70
Q

What is preferred for OBG antigens in screening cells?

A

Homozygous expression is preferred for OBG antigens, although it is not always possible.

71
Q

Are all OBG antigens expressed in the homozygous state?

A

No, some OBG antigens are not expressed in the homozygous state.

72
Q

Why might an antibody screen fail to detect Anti-Jkb?

A

The screen may not detect it if the patient has a very weak Anti-Jkb.

73
Q

What is an example of dosage in antibody screening related to Jkb?

A

A heterozygous Jk(a+b+) cell has too few Jkb antigens to demonstrate agglutination, showing the concept of dosage.

74
Q

What information is provided on Bio-Rad screening cells?

A

They list the donor unit with a cell number.

75
Q

What is the significance of adding an Rh-negative (rr) cell in screening cells?

A

It helps in detecting antibodies specific to Rh-negative cells.

76
Q

What unique identifier does each set of screening cells have?

A

Each set has a specific Lot number.

77
Q

Which blood group antigens are destroyed by enzymes in screening cells?

A

The Duffy and MNS antigens (shaded area) are destroyed by enzymes.

78
Q

What advantage does a 3-cell screen provide in antibody screening?

A

It offers a better chance of detecting all antigens in the homozygous state.

79
Q

For which methods is the 3-cell screen generally used?

A

It is typically used for manual methods rather than automated methods, which often use 2-cell screens.

80
Q

What are the three methods for antibody screening?

A

1) Tube Method, 2) Micro Typing System (MTS or Gel column method), and 3) Solid Phase Red Cell Adherence.

81
Q

How is an antibody screen reported?

A

An antibody screen is reported as either POSITIVE or NEGATIVE.

82
Q

What does a positive antibody screen indicate?

A

It indicates that an antibody was detected.

83
Q

Does the number of screening cells that reacted matter in a positive antibody screen?

A

No, it does not matter how many screening cells reacted for a positive result.

84
Q

Does the method used or the phase in which the reaction occurred matter in a positive antibody screen?

A

No, it does not matter which method was used or which phase it reacted in for a positive result.

85
Q

What does a negative antibody screen indicate?

A

It indicates that no antibody was detected.

86
Q

Does a negative antibody screen mean there is absolutely no antibody present?

A

No, a negative screen does not necessarily mean no antibody is present; a weak antibody might not react with a heterozygous cell.

87
Q

Why can weak antibodies be difficult to detect?

A

Weak antibodies have fewer IgG molecules, making them less likely to react in the screen unless re-stimulated.

88
Q

What happens if a weak antibody is exposed a second time?

A

The immune system may be re-stimulated, leading to increased production of the antibody.

89
Q

What is one limitation of pretransfusion testing?

A

The inability to detect weak antibodies that have not been re-stimulated.

90
Q

When might weak antibodies become detectable?

A

They may become detectable after the patient is re-stimulated, such as after being transfused.

91
Q

Do different antibody screening methods detect different types of antibodies?

A

Yes, different methods detect different types of antibodies.

92
Q

Which phase does the Tube Method include, and what type of antibodies can it detect?

A

The Tube Method includes the Immediate Spin (IS) phase, where cold IgM antibodies can be detected.

93
Q

Which phases do MTS and Capture R include, and what type of antibodies do they detect?

A

MTS and Capture R include only the AHG phase, detecting warm and IgG antibodies, which are considered significant.

94
Q

Do screening cells typically express rare antigens?

A

No, screening cells do not usually express rare antigens, such as Cw+, and will not detect rare antibodies.

95
Q

What must be done when antibodies are detected in a positive screen?

A

All detected antibodies must be identified, regardless of significance.

96
Q

Why is it important to identify all antibodies in a positive screen, even if some are insignificant?

A

An insignificant antibody may be masking another, more important antibody.

97
Q

How many cells are typically found in screening cells compared to panel cells?

A

Screening cells usually have 2-3 cells, while panel cells can have 10-11 cells.

98
Q

How can positive cells (agglutination) be useful in a positive antibody screen?

A

Positive cells (agglutination) can help identify a pattern of the antibody or antibodies.

99
Q

What is the initial setup for the Tube Method in antibody screening?

A

Use 1 drop of 3-5% red cell suspension with 2 drops of patient plasma.

100
Q

What is the first step after adding patient plasma in the Tube Method?

A

Spin and read the results, recording at IS (Immediate Spin) and RT (Room Temperature).

101
Q

What is the incubation temperature and time for the Tube Method?

A

Incubate at 37°C for 15-30 minutes.

102
Q

What steps follow the incubation in the Tube Method?

A

Spin and read the results again.

103
Q

How many washes are required before adding AHG in the Tube Method?

A

Wash three times, then add AHG and read (checking under a microscope if needed).

104
Q

How do you confirm the validity of a negative result in the Tube Method?

A

Check negatives with CCC (Coombs Control Cells) to confirm validity; if testing is valid, mark with a check.

105
Q

What is the purpose of including the Immediate Spin (IS) phase in the Tube Method?

A

Including the IS phase helps detect cold antibodies

106
Q

Are cold antibodies detected in the IS phase usually significant?

A

They are generally insignificant but may mask other antibodies.

107
Q

What is the Tube Method useful for investigating?

A

It is used to investigate anomalies, such as extra reactions.

108
Q

What type of gel is used in the MTS Method?

A

Dextran Acrylamide Gel Spheres.

109
Q

What component is found in both the gel and the diluent in the MTS Method?

A

LISS (Low Ionic Strength Solution).

110
Q

What reagent is used in the MTS Method for antibody screening?

A

Antiglobulin Reagent (AHG).

111
Q

What surrounds the gel spheres in the MTS Method?

A

AHG (Antihuman Globulin).

112
Q

What concentration of red cell suspension is used in the MTS Method?

A

A 0.8% suspension of red cells to maintain the antigen-antibody ratio.

113
Q

What is the first step in the MTS Method antibody screening process?

A

Add test reactants: 50 µL of 0.8% red cell suspension of screening cells I and II to each microtube, and 25 µL of test serum or plasma to each microtube containing screening cells.

114
Q

At what temperature and for how long is the gel card incubated in the MTS Method?

A

Incubate at 37°C for 15 minutes.

115
Q

What is the reaction step in the MTS Method?

A

Centrifuge the gel card for 10 minutes.

116
Q

In the MTS Method, where should red cells ideally stay in the gel column?

A

Red cells should stay at the wider top of the column.

117
Q

What happens to cells coated with antibodies in the MTS Method?

A

Cells coated with antibodies will react with the Anti-Human Globulin (AHG) as they fall through the gel.

118
Q

In the MTS Method, where do un-agglutinated red cells settle in the microtube?

A

Un-agglutinated red cells settle to the bottom of the microtube.

119
Q

In the MTS Method, where are agglutinated red cells found?

A

Agglutinated red cells are trapped in the upper levels of the gel.

120
Q

What does the position of red cells in the MTS microtube indicate?

A

The position indicates whether the cells are agglutinated (trapped at the top) or un-agglutinated (settling at the bottom).

121
Q

What can cause a mixed field reaction due to cold antibody agglutination?

A

A strong cold antibody, like Anti-I, that starts to agglutinate immediately, causing cells to agglutinate at the top where it is coldest, while other cells fall to the bottom.

122
Q

How might technique contribute to a mixed field reaction?

A

If red cells start falling before plasma is added, it could cause a mixed field reaction due to improper technique.

123
Q

What role can fibrin in plasma play in a mixed field reaction?

A

Fibrin in older plasma samples may bind to RBCs at the top, contributing to a mixed field reaction.

124
Q

What is the main principle difference between Capture R Testing and agglutination reactions?

A

Capture R Testing uses a different principle from agglutination reactions, although it still involves antigen and antibody reactions.

125
Q

For which type of antibodies is Capture R Testing exclusively used?

A

Capture R Testing is only used for IgG antibodies.

126
Q

In Capture R Testing, where are red cells (target antigens) located?

A

Red cells are bound to the bottom of the microplate wells.

127
Q

What is the role of protein in Capture R Testing?

A

Protein is bound irreversibly to polystyrene in microplate wells to hold antigens in place.

128
Q

What happens to red cells in Capture R Testing?

A

Red cells are lysed, and their antigens remain bound to the wells.

129
Q

How are plates used in Capture R Testing pre-prepared?

A

Plates come preloaded with RBC antigens.

130
Q

What is the first step in the Capture R test procedure?

A

LISS is added to the microplate wells.

131
Q

What is added to the microplates after LISS in the Capture R test procedure?

A

Patient serum is added to the microplates with ‘bound’ RBC antigens.

132
Q

At what temperature is the Capture R test incubated?

A

The test is incubated at 37°C.

133
Q

What is the purpose of washing the microtitre plate in the Capture R test procedure?

A

To remove excess immunoglobulins from the plate.

134
Q

What are the possible results observed after centrifugation in the Capture R test?

A

Strong positive, weak positive, or negative results based on the presence of agglutination.

135
Q

What is the purpose of adding indicator cells in the test procedure?

A

To detect the presence of Anti-IgG by binding or tagging it to the RBC.

136
Q

How does Anti-IgG bind to the red blood cell (RBC) in the addition of indicator cells?

A

Anti-IgG binds or is “tagged” to the RBC by the Fc portion.

137
Q

What indicates a positive reaction in Capture R testing?

A

The patient’s IgG reacts with the antigen on the wells, causing indicator cells to stick to plate-bound IgG, and the wells are coated with indicator cells.

138
Q

What does a negative reaction look like in Capture R testing?

A

No antibody or IgG is present on the antigens; indicator cells fail to adhere to the well and form a button after centrifugation.

139
Q

In Capture R testing, what happens to indicator cells in a positive reaction?

A

Indicator cells stick to plate-bound IgG, coating the wells.

140
Q

In Capture R testing, what visual result represents a strong positive, weak positive, and negative reaction?

A

A strong positive shows a dark coating, a weak positive shows a lighter coating, and a negative shows a button formation.

141
Q

The Antibody Screen:
1) Detects most clinically significant Antibodies
2) Detects low-frequency antibodies
3) Helps to distinguish between alloantibodies and autoantibodies
4) Can be omitted if the patient has no history of antibodies

A

Detects most clinically significant antibodies

142
Q

An Antibody demonstrating dosage means:
1) Homozygous cells had a stronger reaction
2) Heterozygous cells had a stronger reaction
3) Reacts better with PEG
4) Reacts Best at 4 ˚C

A

Homozygous cells were stronger

143
Q

Saline in an automated cell washer did not fill the test tubes correctly or consistently, used for tube method screening. This was evaluated during evening QC.
1) Would this affect the results of AHG testing?
2) How would this problem be detected in testing?

A

YES, it would affect the results, possible false negative reactions due to unbound antibodies
floating around in suspension, AHG will be neutralized, and after adding Coombs cells, the
reaction will be negative and would indicate an invalid test

144
Q

You added IgG-sensitized red cells to the negative indirect antiglobulin test result of an antibody screen procedure. You observe agglutination. Is the antibody screen interpretation
Positive or Negative? Please explain.

A

The antibody screen procedure is negative. Adding Coombs to a negative result is required and validates the testing; the results should be positive (agglutination) after the addition of
Coombs/Check cells

145
Q

Group O cells are used as a source for commercial screening cells because:
1) Anti-A is detected using O cells
2) Anti-D is detected using O cells
3) Weak subgroups of A react with O cells
4) ABO antibodies do not react with group O cells

A

ABO antibodies do not react with the group O cells

146
Q

What reagent would be selected to detect the presence of unexpected red cell antibodies in a patient’s serum sample?
1) A1 cells
2) B cells
3) IgG sensitized cells
4) Screening cells

A

Screening cells

147
Q

To determine the specificity of a red cell antigen in a patient sample, what antibody source is selected?
1) Commercial reagent red cells
2) Commercial antisera
3) Patient plasma
4) Patient red cell suspensions

A

Commercial Antisera

148
Q

What reagents are derived from plant extracts?
1) Panel Cells
2) Commercial Anti-b
3) Lectins
4) AHG reagents

A

Lectins

149
Q

In the gel test, what is the button of cells at the bottom of the
well called?
1) 4+ positive reaction
2) 1+ positive reaction
3) Negative reaction
4) Invalid reaction

A

Negative reaction

150
Q

What are the indicator cells used in SPRCS assays?
1) IgM-coated red cells used in determining the presence of ABO antibodies
2) Cells used to agglutinate the IgG antibodies to form a pellet
3) IgG-coated red cells that crosslink with IgG antibodies attached to the well
4) Check cells that are added to negative reactions

A

IgG-coated red cells that crosslink with IgG antibodies attached to the well

151
Q

Where can false negative reactions occur in gel and solid-phase testing?
1) Failing to centrifuge
2) Not mixing reagent red cells sufficiently
3) Incubation time was too short
4) All of the above

A

All of the above

Transfusion (8 decks)

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