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. 9
Rh Typing (Tube Method)

I. Introduction
Rh typing tube method is used to confirm slide testing. Rh antigen typing
uses anti-D commercially prepared sera. It must be performed with strict
adherence to manufacturer’s directions. Proper interpretation of test results is
essential since most inaccurate or incorrect results of Rh typing occur due to
technical errors. These can result from defects in equipment, reagents, specimen
and techniques or through wrong interpretation.

II. Objectives
At the end of the activity, the student must be able to:
1. perform Rh typing using tube method
2. give the possible genotypes of Rh positive and Rh negative blood.
3. Know the importance of Rh control.

III. Materials
Test Tubes
Test Tube Rack
Parafilm

IV. Reagents/ Samples

Normal Saline Solution (NSS)
Patient’s Blood at EDTA tube
Anti-Rh (Anti-D typing sera)

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

V. Procedure
1. Prepare 5% RCS of patient’s blood.
2. Label two test tubes with U (Unknown) and NC (Negative Control).
3. Place 1 drop of 5% RCS of patient’s blood into each tube.
4. Add 1 drop of Anti-D to tube labelled “U” and 1 drop of 22% Bovine Albumin to
the labelled “NC”.
5. Mix gently. Cover with parafilm. Centrifuge all tubes for 15-30 seconds.
6. Gently dislodge the cell button and examine for agglutination.
7. Grade each reaction and record the results.

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 (Tube Method)

Name: _________________________________________ Date: __________________

Procedure Rating Comments

1. Wear laboratory gown
2. Wash hands and wear gloves
3. Assemble equipment and materials for Rh typing
(Tube Method)
4. Prepare 5% RCS of patient’s blood
5. Label two test tubes with U (Unknown) and NC
(Negative Control).
6. Place 1 drop of 5% RCS of patient’s blood into each
tube.
7. Add 1 drop of Anti-D to tube labelled “U” and 1 drop of
22% Bovine Albumin to the labelled “NC”.
8. Mix gently. Cover with parafilm. Centrifuge all tubes for
15-30 seconds.
9. Gently dislodge the cell button and examine for
agglutination.
10. Grade each reaction and record the results.
12. Clean and return the equipment to proper storage.
13. Clean the work area with surface disinfectant.
14. 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. 9
Rh Typing (Tube Method)

Name: Rating:
Date Performed: Date Submitted:

Results:
Illustrate your final result obtained.

Unknown Negative Control

Reaction: _____________ 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. Give possible genotypes of Rh-positive blood.

a.
b.
c.
d.
e.

2. Give the possible genotypes of Rh-negative blood.

a.
b.
c.
d.
e.

3. Can reverse typing be done to detect the Rh type of an individual? Why?

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

4. If a person is transfused with an Rh (-) blood for the first time, what will be the
immunological consequences to this patient? What will be the outcome of the first
transfusion?

5. Name the common reagent that serve as control for Rh typing and give the
significance of this control.

Laboratory Manual in Immunohematology (MT 313)

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