BB PRELIMS LAB

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RBC Suspension, ABO, Rh

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104 Terms

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Whole blood (Anticoagulated, preferably EDTA)

Specimen requirement for red cell suspension

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term image

Formula for red cell suspension

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To remove unbound antibodies and excess proteins

Purpose of washing RBCs

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tomato red or cherry red color

Color of proper 5% red cell suspension

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2-5% RBC Suspension

What is the percent of RBC suspension that is the universally accepted proportion of blood for most manual blood bank procedures?

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Landsteiner’s Rule

The ABO blood group system is the only system in which the reciprocal antibodies are consistently and predictably present in the sera of normal people whose RBCs lack the corresponding antigen(s). This rule is known as _______?

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Anti-B

Antibody/Antibodies Present in Blood Group A

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Anti-A

Antibody/Antibodies Present in Blood Group B

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None

Antibody/Antibodies Present in Blood Group AB

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Anti-A, Anti-B, and Anti-AB

Antibody/Antibodies Present in Blood Group O

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A

Antigen(s) Present in Blood Group A

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B

Antigen(s) Present in Blood Group B

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A and B

Antigen(s) Present in Blood Group AB

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None

Antigen(s) Present in Blood Group O

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Fill in the Table

<p>Fill in the Table</p>
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Forward Typing

Process of testing unknown cells with known antibodies:

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Reverse Typing

Process of testing an unknown serum/plasma with known RBCs

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Red Cells

Sample used for Forward Typing

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Serum/Plasma

Sample used for Reverse Typing

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Anti-Sera or Known Antibodies

Reagent used for Forward Typing

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Known RBCs

Reagent used for Reverse Typing

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ABO Forward and Reverse Grouping Rest 

Most frequently performed tests in the blood bank.

It must be performed on all donors and blood transfusion recipients.

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ABO antigens are detected on the red cells of the embryo as early as five weeks after conception.

ABO antigens are detected on the red cells of the embryo as early as?

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ABO antigens develop as early as the 37th week of fetal life.

ABO antigens develop as early as?

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Fetal and Newborn Infant’s RBCs

Weaker reactions (agglutination) are seen in this age group as their RBCs have less antigenic sites

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Expression of ABO Antigens is fully developed by 2 to 4 years of age.

Expression of ABO Antigens is fully developed by?

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Forward (Front) / Direct / Cell Typing

This test is used to detect the presence of ABO antigens on the red cells of the unknown blood sample by mixing the red cells of the unknown blood sample with commercially prepared antisera.

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Blue

Color of Anti-A antisera

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Yellow

Color of Anti-B antisera

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term image

Fill in the Table

<p>Fill in the Table</p>
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The formation of antibodies occurs in the first few months of life.

The formation of antibodies occurs in?

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Antibody titers are low up to 3-6 months of age.

Antibody titers are low up to _____ months of age.

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Reverse typing is not applicable for patients less than 6 months of age because most or all antibodies present are IgGs from maternal origin, and antibodies are generally too low for detection.

Reverse typing is not applicable for patients __________ of age.

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Antibodies peak at 5 to 10 years of age, remain constant and decline later in life

Antibodies peak at _______ years of age

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Antibodies may be very low or undetectable in reverse typing of serum from elderly persons.

Antibodies may be ___________ in reverse typing of serum from elderly persons.

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Reverse (Back) / Indirect / Serum Typing

The test is used to confirm the result of the forward typing by detecting the presence of antibodies in the serum of the patient.

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Reverse (Back) / Indirect / Serum Typing

It is based on the principle that the serum contains antibodies towards the antigens lacking on the red cells of the unknown blood sample.

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Known-A and Known-B RBC suspensions

In Reverse (Back) / Indirect / Serum Typing, the serum of the patient is tested with?

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term image

Fill in the Table

<p>Fill in the Table</p>
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Slide Method

[Methods in Blood Typing]

Rudimentary method, not recommended for transfusion purposes.

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Slide Method

[Methods in Blood Typing]

Commercially prepared reagent and unknown blood are mixed on a slide with an applicator stick. 

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In the slide method, longer than 2 minutes of incubation should be avoided because of the effects of drying that may be misinterpreted as agglutination (pseudoagglutination or false positive).

In the slide method, longer than ________ of incubation should be avoided because of the effects of drying that may be misinterpreted as agglutination (pseudoagglutination or false positive).

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Tube Method

[Methods in Blood Typing]

Follows the same principle as the slide method, but the samples and reagents are placed on a clean test tube, then centrifuged and examined both macroscopically and microscopically for agglutination. 

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True

[True or False]

The tube method is more sensitive than the slide method and may be used for transfusion purposes.

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Gel Method

[Methods in Blood Typing]

Detects antigen-antibody reaction with the use of a polyacrylamide gel, which acts as a trap that sequesters agglutinated cells.

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Agglutination is observed as RBCs trapped within the gel matrix, whereas unagglutinated cells appear as forming a pellet at the bottom of the microtube.

Agglutinated: Trapped within the Gel Matrix (Above)

Unagglutinated: Bottom of the Microtube

[Methods in Blood Typing]

Agglutination is observed as RBCs trapped within the gel matrix, whereas unagglutinated cells appear as forming a pellet at the ______ of the microtube.

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Grouping reagents (anti-sera) must be tested daily to confirm the specificity of red cells and check the specificity and avidity of reagents. 

Grouping reagents (anti-sera) must be tested _____ to confirm the specificity of red cells and check the specificity and avidity of reagents. 

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4+

[GRADING OF REACTION IN TUBE METHOD]

one solid agglutinate, no free cells, clear supernatant fluid (macroscopic)

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3+

[GRADING OF REACTION IN TUBE METHOD]

several large agglutinates, few free cells, clear supernatant (macroscopic)

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2+

[GRADING OF REACTION IN TUBE METHOD]

medium-sized agglutinates, some free cells, clear supernatant (macroscopic)

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1+

[GRADING OF REACTION IN TUBE METHOD]

small agglutinates, many free cells, turbid background, reddish supernatant

(macroscopic and microscopic reading)

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0

[GRADING OF REACTION IN TUBE METHOD]

Negative, no agglutination, smooth suspension of cells.

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1w

[GRADING OF REACTION IN TUBE METHOD]

very small/tiny agglutinates (microscopic), turbid background, reddish supernatant (microscopic)

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MF

[GRADING OF REACTION IN TUBE METHOD]

mixed-field, few isolated agglutinates with large areas of unagglutinated cells, reddish supernatant (microscopic)

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H

[GRADING OF REACTION IN TUBE METHOD]

Complete Hemolysis

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PH

 [GRADING OF REACTION IN TUBE METHOD]

Partial hemolysis, some RBCs remain

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Forward/Cell Typing (Slide and Tube Method)

The principle of this method is the detection of antigen(s) on RBCs using known sources of commercial antisera (anti-A and anti-B).

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Anticoagulated blood / whole blood / RBC cell suspension

Specimen requirement for Forward or Cell Typing Slide Method

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Presence of Agglutination 

Positive Result in Forward Typing Slide Method

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No Agglutination 

Negative Result in Forward Typing Slide Method

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<ul><li><p><strong>Blood Group: </strong>A</p></li><li><p><strong>Antibody/Antibodies Present: </strong>Anti-B</p></li><li><p><strong>Antigen(s) Present:</strong> A</p></li></ul><p></p>

  • Blood Group: A

  • Antibody/Antibodies Present: Anti-B

  • Antigen(s) Present: A

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p>
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<ul><li><p><strong>Blood Group: </strong>B</p></li><li><p><strong>Antibody/Antibodies Present: </strong>Anti-A</p></li><li><p><strong>Antigen(s) Present:</strong> B</p></li></ul><p></p>

  • Blood Group: B

  • Antibody/Antibodies Present: Anti-A

  • Antigen(s) Present: B

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p>
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<ul><li><p><strong>Blood Group: </strong>AB</p></li><li><p><strong>Antibody/Antibodies Present: </strong>None</p></li><li><p><strong>Antigen(s) Present:</strong> A and B</p></li></ul><p></p>

  • Blood Group: AB

  • Antibody/Antibodies Present: None

  • Antigen(s) Present: A and B

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p>
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<ul><li><p><strong>Blood Group: </strong>O</p></li><li><p><strong>Antibody/Antibodies Present: </strong>Anti-A, Anti-B, and Anti-AB</p></li><li><p><strong>Antigen(s) Present:</strong> None</p></li></ul><p></p>

  • Blood Group: O

  • Antibody/Antibodies Present: Anti-A, Anti-B, and Anti-AB

  • Antigen(s) Present: None

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p>
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2-5% patient RBC suspension

Specimen requirement for Forward or Cell Typing Tube Method

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Presence of Agglutination 

Positive Result in Forward Typing Tube Method

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No Agglutination

Negative Result in Forward Typing Tube Method

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<ul><li><p><strong>Blood Group:</strong> A</p></li></ul><ul><li><p><strong>Antibody/Antibodies Present:</strong> Anti-B</p></li><li><p><strong>Antigen(s) Present:</strong> A</p></li></ul><p></p><ul><li><p><strong>Anti-A Sera: </strong>4+</p></li><li><p><strong>Anti-B Sera:</strong> 0</p></li></ul><p></p>

  • Blood Group: A

  • Antibody/Antibodies Present: Anti-B

  • Antigen(s) Present: A

  • Anti-A Sera: 4+

  • Anti-B Sera: 0

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

Grade the Reaction

  • Anti-A Sera:

  • Anti-B Sera:

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p><p>Grade the Reaction</p><ul><li><p>Anti-A Sera: </p></li><li><p>Anti-B Sera:</p></li></ul><p></p>
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<ul><li><p><strong>Blood Group:</strong> B</p></li></ul><ul><li><p><strong>Antibody/Antibodies Present:</strong> Anti-A</p></li><li><p><strong>Antigen(s) Present:</strong> B</p></li></ul><p></p><ul><li><p><strong>Anti-A Sera: </strong>0</p></li><li><p><strong>Anti-B Sera:</strong> 4+</p></li></ul><p></p>

  • Blood Group: B

  • Antibody/Antibodies Present: Anti-A

  • Antigen(s) Present: B

  • Anti-A Sera: 0

  • Anti-B Sera: 4+

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

Grade the Reaction

  • Anti-A Sera:

  • Anti-B Sera:

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p><p>Grade the Reaction</p><ul><li><p>Anti-A Sera:</p></li><li><p>Anti-B Sera:</p></li></ul><p></p>
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<ul><li><p><strong>Blood Group:</strong> AB</p></li></ul><ul><li><p><strong>Antibody/Antibodies Present:</strong> None</p></li><li><p><strong>Antigen(s) Present:</strong> A and B</p></li></ul><p></p><ul><li><p><strong>Anti-A Sera: </strong>4+</p></li><li><p><strong>Anti-B Sera:</strong> 4+</p></li></ul><p></p>

  • Blood Group: AB

  • Antibody/Antibodies Present: None

  • Antigen(s) Present: A and B

  • Anti-A Sera: 4+

  • Anti-B Sera: 4+

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

Grade the Reaction

  • Anti-A Sera:

  • Anti-B Sera:

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p><p>Grade the Reaction</p><ul><li><p>Anti-A Sera: </p></li><li><p>Anti-B Sera:</p></li></ul><p></p>
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<ul><li><p><strong>Blood Group:</strong> O</p></li></ul><ul><li><p><strong>Antibody/Antibodies Present:</strong> Anti-A, Anti-B, and Anti-AB</p></li><li><p><strong>Antigen(s) Present:</strong> None</p></li></ul><p></p><ul><li><p><strong>Anti-A Sera: </strong>0</p></li><li><p><strong>Anti-B Sera:</strong> 0</p></li></ul><p></p>

  • Blood Group: O

  • Antibody/Antibodies Present: Anti-A, Anti-B, and Anti-AB

  • Antigen(s) Present: None

  • Anti-A Sera: 0

  • Anti-B Sera: 0

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

Grade the Reaction

  • Anti-A Sera:

  • Anti-B Sera:

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p><p>Grade the Reaction</p><ul><li><p>Anti-A Sera: </p></li><li><p>Anti-B Sera:</p></li></ul><p></p>
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Reverse/Serum Typing (Slide and Tube Method)

The principle of this process is the detection of ABO antibody (ies) in the patient’s serum by using known RBC suspension, namely; A1 and B cells

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<ul><li><p><strong>Blood Group: </strong>A</p></li><li><p><strong>Antibody/Antibodies Present: </strong>Anti-B</p></li><li><p><strong>Antigen(s) Present: </strong>A</p></li></ul><p></p>

  • Blood Group: A

  • Antibody/Antibodies Present: Anti-B

  • Antigen(s) Present: A

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p>
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<ul><li><p><strong>Blood Group: </strong>B</p></li><li><p><strong>Antibody/Antibodies Present: </strong>Anti-A</p></li><li><p><strong>Antigen(s) Present: </strong>B</p></li></ul><p></p>

  • Blood Group: B

  • Antibody/Antibodies Present: Anti-A

  • Antigen(s) Present: B

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p>
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<ul><li><p><strong>Blood Group: </strong>AB</p></li><li><p><strong>Antibody/Antibodies Present: </strong>None</p></li><li><p><strong>Antigen(s) Present: </strong>A and B</p></li></ul><p></p>

  • Blood Group: AB

  • Antibody/Antibodies Present: None

  • Antigen(s) Present: A and B

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p>
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<ul><li><p><strong>Blood Group:</strong> O</p></li><li><p><strong>Antibody/Antibodies Present: </strong>Anti-A, Anti-B, and Anti-AB</p></li><li><p><strong>Antigen(s) Present:</strong> None</p></li></ul><p></p>

  • Blood Group: O

  • Antibody/Antibodies Present: Anti-A, Anti-B, and Anti-AB

  • Antigen(s) Present: None

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p>
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<ul><li><p><strong>Blood Group: </strong>A</p></li></ul><ul><li><p><strong>Antibody/Antibodies Present: </strong>Anti-B</p></li><li><p><strong>Antigen(s) Present:</strong> A</p></li></ul><p></p><ul><li><p><strong>Known A1:</strong> 0</p></li><li><p><strong>Known B: </strong>4+</p></li></ul><p></p>

  • Blood Group: A

  • Antibody/Antibodies Present: Anti-B

  • Antigen(s) Present: A

  • Known A1: 0

  • Known B: 4+

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

Grade the Reaction

  • Known A1:

  • Known B:

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p><p>Grade the Reaction</p><ul><li><p>Known A1:</p></li><li><p>Known B:</p></li></ul><p></p>
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<ul><li><p><strong>Blood Group: </strong>B</p></li></ul><ul><li><p><strong>Antibody/Antibodies Present: </strong>Anti-A</p></li><li><p><strong>Antigen(s) Present: </strong>B</p></li></ul><p></p><ul><li><p><strong>Known A1: </strong>4+</p></li><li><p><strong>Known B: </strong>0</p></li></ul><p></p>

  • Blood Group: B

  • Antibody/Antibodies Present: Anti-A

  • Antigen(s) Present: B

  • Known A1: 4+

  • Known B: 0

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

Grade the Reaction

  • Known A1:

  • Known B:

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p><p>Grade the Reaction</p><ul><li><p>Known A1:</p></li><li><p>Known B:</p></li></ul><p></p>
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<ul><li><p><strong>Blood Group: </strong>O</p></li></ul><ul><li><p><strong>Antibody/Antibodies Present: </strong>Anti-A, Anti-B, and Anti-AB</p></li><li><p><strong>Antigen(s) Present: </strong>None</p></li></ul><p></p><ul><li><p><strong>Known A1: </strong>4+</p></li><li><p><strong>Known B: </strong>4+</p></li></ul><p></p>

  • Blood Group: O

  • Antibody/Antibodies Present: Anti-A, Anti-B, and Anti-AB

  • Antigen(s) Present: None

  • Known A1: 4+

  • Known B: 4+

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

Grade the Reaction

  • Known A1:

  • Known B:

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p><p>Grade the Reaction</p><ul><li><p>Known A1:</p></li><li><p>Known B:</p></li></ul><p></p>
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<ul><li><p><strong>Blood Group: </strong>AB</p></li></ul><ul><li><p><strong>Antibody/Antibodies Present: </strong>None</p></li><li><p><strong>Antigen(s) Present: </strong>A and B</p></li></ul><p></p><ul><li><p><strong>Known A1: </strong>0</p></li><li><p><strong>Known B: </strong>0</p></li></ul><p></p>

  • Blood Group: AB

  • Antibody/Antibodies Present: None

  • Antigen(s) Present: A and B

  • Known A1: 0

  • Known B: 0

Identify the:

  • Blood Group

  • Antibody/Antibodies Present

  • Antigen(s) Present

Grade the Reaction

  • Known A1:

  • Known B:

<p>Identify the:</p><ul><li><p>Blood Group</p></li><li><p>Antibody/Antibodies Present</p></li><li><p>Antigen(s) Present</p></li></ul><p></p><p>Grade the Reaction</p><ul><li><p>Known A1:</p></li><li><p>Known B:</p></li></ul><p></p>
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Rh is the most important blood group system after ABO in transfusion medicine.

___ is the most important blood group system after ABO in transfusion medicine.

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D antigen

Most potent Rh antigen

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Most Rh antibodies are IgG immunoglobulins, and they react best at 37ºC

Most Rh antibodies are ___ immunoglobulins and they react best at __ºC

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Presence of agglutination in test, negative in control

Positive Reaction for Rh Testing

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No agglutination on both test and control

Negative Reaction for Rh Testing

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D, C, E, c, e

There are five (5) available reagent antisera to test for the five common Rh antigens:

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Anti-D 

The only Rh required for routine testing.

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Rh-D Typing (Slide and Tube Method)

The principle of this test is the detection of the presence of the Rh D antigen using commercially available anti-D typing sera.

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Whole blood / anticoagulated blood (EDTA) / washed RBC suspension

Specimen used in Rh Typing (Slide and Tube Method)

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Anti-D typing sera

Reagent used in Rh Typing Slide Method

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Bovine albumin or 22% LISS 

Used as Control in Rh Test

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<p>4+</p><p>one solid agglutinate, no free cells, clear supernatant fluid (macroscopic)</p>

4+

one solid agglutinate, no free cells, clear supernatant fluid (macroscopic)

[GRADE THE REACTION]

<p><strong>[GRADE THE REACTION]</strong></p>
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<p>3+</p><p>several large agglutinates, few free cells, clear supernatant (macroscopic)</p>

3+

several large agglutinates, few free cells, clear supernatant (macroscopic)

[GRADE THE REACTION]

<p><strong>[GRADE THE REACTION]</strong></p>
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<p>2+</p><p>medium-sized agglutinates, some free cells, clear supernatant (macroscopic)</p>

2+

medium-sized agglutinates, some free cells, clear supernatant (macroscopic)

[GRADE THE REACTION]

<p><strong>[GRADE THE REACTION]</strong></p>
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<p>1+</p><p>small agglutinates, many free cells, turbid background, reddish supernatant (macroscopic and microscopic reading)</p>

1+

small agglutinates, many free cells, turbid background, reddish supernatant (macroscopic and microscopic reading)

[GRADE THE REACTION]

<p><strong>[GRADE THE REACTION]</strong></p>
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<p>Hemolysis</p><p>complete hemolysis</p>

Hemolysis

complete hemolysis

[GRADE THE REACTION]

<p><strong>[GRADE THE REACTION]</strong></p>
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<p>Negative Reaction</p><p>Negative, no agglutination, smooth suspension of cells.</p>

Negative Reaction

Negative, no agglutination, smooth suspension of cells.

[GRADE THE REACTION]

<p><strong>[GRADE THE REACTION]</strong></p>
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<p>4+</p>

4+

[GRADE THE REACTION]

<p><strong>[GRADE THE REACTION]</strong></p>