ST.

ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com

Activity No. 8
Rh Typing (Slide Method)

I. Introduction
The first and most clinically important characterization of Rh (D) system
antigens came when Landsteiner and Weiner (1940) published studies of animal
experiments involving the immunization of guinea pigs and rabbit with Rhesus
monkey’ s RBCs. The resulting antiserum agglutinated 85% of human RBCs and
the antigen defined was called the Rh (rhesus) factor.
Rh antigens are highly immunogenic. The Rh (D) antigen is the most
immunogenic antigen outside ABO system. It is important to test D antigen in all
patients who are to receive blood transfusion, so that proper type of blood will be
given. Rh negative patients should be transfused with Rh Negative blood.
Testing for D antigen is also a way of identifying females who might be at risk of
giving birth to an infant with Hemolytic Disease of Newborn (HDN). The five
major Rh antigens in decreasing immunogenicity are as follows: D, c, E, C, e.

II. Objectives
At the end of the activity, the student must be able to:
1. perform Rh blood typing using the slide method.
2. classify the red cells depending on the presence of absence of D antigen.
3. determine the importance of D antigen testing.
4. identify the possible false positive reactions with Rh typing.

III. Materials
Applicator Stick
Glass slide

Laboratory Manual in Immunohematology (MT 313)

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 ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com

IV. Reagents/ Sample

Patient’s Blood in EDTA tube
Rh Typing Sera (Anti-D)
22% Bovine Albumin

V. Procedure
1. Prepare 2 glass slides. Label on one slide “Test” and the other “Control.
2. Place 1 drop of Anti-D on the slide marked “Test”
3. Place 1 drop of 22 % Bovine Albumin on the slide marked “Control”.
4. To each slide, add 1 drop of patient’s blood.
5. With separate applicator sticks, mix each preparation and spread over an
area of about 2-4 cm.
6. Tilt back and forth and observe for agglutination within 2 minutes.
7. Agglutination indicates positive result.

Laboratory Manual in Immunohematology (MT 313)

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 ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com

PERFORMANCE ASSESSMENT SHEET

Rh Typing (Slide Method)

Name: _________________________________________ Date: __________________

Procedure Rating Comments

1. Wear laboratory gown
2. Wash hands and wear gloves
3. Assemble equipment and materials for Rh typing
(Slide Method)
4. Prepare 2 glass slides. Label on one slide “Test” and
the other “Control.
5. Place 1 drop of Anti-D on the slide marked “Test”
6. Place 1 drop of 22 % Bovine Albumin on the slide
marked “Control”.
7. To each slide, add 1 drop of patient’s blood.
8. With separate applicator sticks, mix each preparation
and spread over an area of about 2-4 cm.
9. Tilt back and forth and observe for agglutination within
2 minutes.
10. Clean and return the equipment to proper storage.
11. Clean the work area with surface disinfectant.
12. Safely disposed the gloves in a pathologic waste
container and properly washed hands with disinfectant.

Rating: 4= Excellent 3= Very Satisfactory 2= Satisfactory 1=Poor

Level of Competency:

Instructor: Date:

Laboratory Manual in Immunohematology (MT 313)

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 ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com

REPORT SHEET
Activity No. 8
Rh Typing (Slide Method)

Name: Rating:
Date Performed: Date Submitted:

Results:
Illustrate your final result obtained.

Test

Reaction: _________________

Control

Reaction: __________________

Laboratory Manual in Immunohematology (MT 313)

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 ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com

Review Questions:
1. What are the importance of Rh typing?
a.
b.
c.
d.
e.
f.

2. Enumerate the false positive and false negative reactions in Rh typing and give the
counterpart corrective actions to each.

3. How is Rh typing serum prepared?

Laboratory Manual in Immunohematology (MT 313)

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 ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com

Laboratory Manual in Immunohematology (MT 313)

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