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# Determining the ABO-Rh Blood Type

Determining the ABO-Rh blood type

of simulated samples

part 1: objectives/aims

The aim of this experiment is to determine the blood type of Patient 2 using the Edu-Lab ABO-Rh Blood typing kit and blood type of individual (self) using the EldonCard 2521.

PART 2: INTRODUCTION

There are 8 blood groups and they are classified using two systems:

     ABO System

     Rhesus System

ABO System

myDr, 2016 states that two of the antigens found on the surface of red blood cells are called antigen A and antigen B. The ABO group system for blood typing is based on which of these antigens a person has on their red blood cells. A person will inherit the particular combination of blood group antigens they have from their parents.

According to myDr, 2016, the first system is called the ABO system and it has four main groups:

1. A Group

Figure 1    Showing the ABO system of the four blood types

• Blood group A have A antigens on the surface of the red blood cells and B antibodies in the blood plasma as seen in Figure 1.
• Patients with A blood type can donate A and AB blood types
• Patients with A blood type can receive A and O blood types

(Austin Community College, 2018)

1. B Group
• Blood group B have B antigens on the surface of the red blood cells and A antibodies in the blood plasma as seen in Figure 1.
• Patients with B blood type can donate B and AB blood types
• Patients with B blood type can receive B and O blood types
1. AB Group
• Blood group AB both have A and B antigens on the surface of the red blood cells and no A or B antibodies at all in the blood plasma as seen in Figure 1.
• Patients with AB blood type can donate AB blood type only
• Patients with AB blood type can receive A, B, AB and O blood types
1. O Group
• Blood group O have neither A or B antigens on the surface of the red blood cells but have both A and B antibodies in the blood plasma as seen in Figure 1.
• Patients with O blood type can donate A, B, AB and O blood types
• Patients with O blood type can receive O blood type only

Rhesus System

According to myDr, 2016, Rhesus system is the other blood typing system commonly used and its also called Rh system, named after the Rhesus monkey in which it was first discovered. The RhD gene encodes a cell surface protein known as the Rhesus D (RhD) antigen or the Rhesus factor antigen.

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In this system, if you have an RhD antigen on the surface of your red blood cells as seen in Figure 2, you are said to be Rhesus positive (Rh+). Red blood cells that lack the RhD antigen are Rhesus negative (Rh-) which is evident in Figure 3. (myDr, 2016)

Figure 2    Showing the Rhesus Positive       Figure 3    Showing the Rhesus Negative

(Elkin, 2014)         (Elkin, 2014)

A person with Rh- blood does not have Anti-RhD antibodies naturally in the blood plasma. However, as seen in Figure 3, a person with Rh- blood can develop Anti-RhD antibodies in the blood plasma if he or she receives blood from a person with Rh+ blood without any problems. (myDr, 2016)

     If a mother is Rh negative but her baby is Rh positive (which can happen if the father is Rh positive), the mother will produce antibodies that fight the baby’s red blood cells. This can happen if blood from the unborn baby enters the mother’s circulation. (myDr, 2016)

     When there is a risk of threatened miscarriage, termination, chorionic villus sampling (CVS), or abdominal trauma at delivery, an injection called Anti-D can be given to the mother to help prevent these antibodies against Rh positive blood being produced. (myDr, 2016)

According to myDr, 2016, combining your ABO blood group with whether you are Rh+ or Rh- means your blood can be classified as one of 8 possible types as seen in Figure 4:

Figure 4    A representation of blood types showing the different types of antigens and antibodies present on each type of blood cell.

• O positive (O+)
• O negative (O-)
• A positive (A+)
• A negative (A-)
• B positive (B+)
• B negative (B-)
• AB positive (AB+)
• AB negative (AB-)

Therefore, the purpose of this experiment is to check how antigens and antibodies interact and to know the individual blood type. The presence or absence of agglutinates will show the blood type.

(Algonquin College, 2012)

part 3: MATERIALS & Methods

Materials:

▫       Anti-A serum (simulated)

▫       Anti-B serum (simulated)

▫       Anti-Rh serum (simulated)

▫       Blood typing tray

▫       Simulated Patient 2 blood sample

▫       Stirring sticks (blue, green, yellow) – 1 set

▫       EldonCard 2521

▫       Lancet

▫       EldonSticks – 4

▫       Ethanol wipes

Methods:

Patient 2 blood type:

1. 5 drops of Patient 2 simulated blood sample was placed in each well (A, B, Rh0) onto a blood typing tray.
2. 3 drops of Anti-A simulated serum was placed in Well A.
3. 3 drops of Anti-B simulated serum was placed in Well B.
4. 3 drops of Anti-Rh simulated serum was placed in Well Rh.
5. A separate stirring stick was used to mix the simulated blood and serum in each well for about 10 seconds.
6. Each well was carefully examined to determine if the simulated blood in each well has clumped together or agglutinated. The results were recorded in Table 1.

Individual blood type:

1. The data of the person being tested was filled on the EldonCard 2521.
2. Using a micro-dropper, a drop of water was placed onto each of the coloured reagents spots on the EldonCard.
3. The finger of the person being tested was disinfected using an Ethanol wipes until it was dried.
4. The little green protective lancet cap was removed by twisting it off. Afterwards, the lancet was held against the fingertip and the green body was pressed against the finger to release the needle.
5. The fingertip was pressed firmly to release the blood. Using an EldonStick, the blood was transferred into the Anti-A reagent. This procedure was repeated 3 times using the remaining EldonSticks.
6. The blood was stirred for approximately 10 seconds in each reagent spots.
7. Each reagent spot was carefully examined to determine if the blood in each spot has clumped together or agglutinated. The results were read and recorded.

part 4: results

Table 1    Showing the data results of Patient 2 simulated blood sample

 Simulated blood sample Agglutination in Well A (+/-) Agglutination in Well B (+/-) Agglutination in Well Rh (+/-) Blood Type Patient 2 + – + A +

Table 1 shows there was agglutination in Well A and in Well Rh and the corresponding blood type is A+

Figure 5    Showing the results from the data table 1

Figure 5 shows that there was agglutination when the simulated blood of Patient 2 was mixed with Anti-A serum in well A and Anti-Rh serum in well Rhwhile there was no change with Anti-B serum in well B. This represents that Patient 2 carried the blood type of A+.

Table 2    Showing the data results of the Individual blood sample

 Simulated blood sample Agglutination in Anti-A (+/-) Agglutination in Anti-B (+/-) Agglutination in Anti-D (+/-) Agglutination in Control (+/-) Blood Type Individual – – + – O +

Table 2 shows there was agglutination in Anti-D serum and the corresponding blood type is O+

Figure 6    Showing the results from the data table 2

Figure 6 shows that there was agglutination when the blood (from individual) were mixed with Anti-D serum while there was no change in control, Anti-A antibodies and Anti-B antibodies. Figure 6 represents that the individual carried the blood type of O+.

part 5: discussion

The objectives of this experiment were successfully performed. It was possible to determine the blood type of Patient 2 using the Edu-Lab ABO-Rh Blood typing kit and blood type of individual (self) using the EldonCard 2521.

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Based on Table 1 and Figure 5, it showed that there was agglutination when the simulated blood of Patient 2 was mixed with Anti-A serum in well A and Anti-Rh serum in well Rhwhile there was no change with Anti-B serum in well B. Therefore, Patient 2 carried the blood type of A+ because there were A and Rh antigens on the red blood cells and Anti-B antibodies in the blood plasma of Patient 2.

Based on Table 2 and Figure 6, it showed that there was agglutination when the blood (from individual) were mixed with Anti-D serum while there was no change in Anti-A antibodies and Anti-B antibodies. There was also no change in control, therefore this produced a valid experiment because if the control showed agglutination, then it means the EldonCard was contaminated. As a result, the individual carried the blood type of O+ because there was Rh antigens on the red blood cells and Anti-A and Anti-B in the blood plasma of the individual.

To improve the results if the experiment were to repeat again;

• Adding blood (individual) onto the control reagent spot to analyse if there is contamination. No agglutination means no contamination and therefore a valid result.
• Adding more drops of Anti-A, Anti-B and Anti-Rh serum onto the wells to obtain an adequate agglutination for Patient 2 blood sample.
• Observation for Patient 1, 3 and 4 simulated blood types to determine what type of blood they carry and produce precise results.

Questions

ABO

1. What was the ABO type of the blood tested?
• The ABO type of Patient 2 simulated blood was A.
1. What ABO antigens are present in the red blood cells of this type of blood?
• The ABO antigens that are present in blood type A is A antigens.
1. What ABO antibodies are present in the red blood cells?
• The ABO antibodies that are present in blood type A is Anti-B antibodies.
1. If a person with this blood type needed a blood transfusion, what ABO type(s) of blood could be received safely?
• Blood type A can receive from O or A blood types.
1. If a person with this blood type as serving as a blood donor, what ABO blood type(s) could receive the blood safely?
• Blood type A can donate to A or AB blood types.

Rhesus

1. What was the Rh type of the blood tested in the demonstration?
• The Rh type of Patient 2 simulated blood was A+
1. What Rh antigen is present in the red blood cells of this type of blood?
• The Rh antigens that are present in blood type A+ are A antigens and Rh antigens.
1. What Rh antibody is normally present in the plasma of this type of blood?
• The Rh antibodies that are present in blood type A+ is Anti-B antibodies
1. If a person with this blood type needed a blood transfusion, what type of blood could be received safely?
• Blood type A+ can receive from any O or A blood types.
1. If a person with this blood type was serving as a blood donor, a person with what type of blood could receive the blood safely?
• Blood type A+ can donate to A+ or AB+ blood types.

part 6: conclusion

In conclusion, the aim to identify the blood type of Patient 2 and the blood type of individual (self) was achieved by using the Edu-Lab ABO-Rh Blood typing kit and EldonCard 2521. The simulated blood of Patient 2 was identified as A+ and the blood type of the individual was identified as O+. Therefore, this assay/method can determine blood types quickly, easily and reliably.

part 7: references

      Kelly, B. and Pollock, J. (2018). 3rd Year Immunology Semester 1 Lab Manual [unpublished], PHARS7013: Immunology Dundalk Institute of Technology, Department of Applied Science.

      Algonquin College. 2012. Anatomy and Physiology. [ONLINE] Available at: http://lyceum.algonquincollege.com/lts/onlineCourses/anatomy/content/module11-9.htm. [Accessed 14 October 2018].

      myDr. 2016. Blood typing – ABO blood groups and Rh types [ONLINE] Available at: https://www.mydr.com.au/tests-investigations/blood-typing. [Accessed 14 October 2018].

      Elkin, F. 2014. Blood Typing ABO. [ONLINE] Available at: https://slideplayer.com/slide/2299670/. [Accessed 14 October 2018].

      Austin Community College. 2018. Cardiovascular System (Blood). [ONLINE] Available at: http://www.austincc.edu/apreview/PhysText/Blood.htm. [Accessed 14 October 2018].