blood grouping experiment pdf

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Blood Group Test

We all should be aware of our blood groups. It plays a very important role when it comes to having a blood transfusion. Knowing our blood group helps us to select appropriate blood for transfusion either during any emergencies or accidents.

Here is a simple and easy experiment to find out our blood type sitting at home with the help of a Blood Group Test Kit.

A human blood cell includes red blood cells, white blood cells, and platelets. All these blood cells play a significant role in transportation, protection and regulation. Blood groups are mainly determined by the presence or absence of antigens and antibodies on the surface of our red blood cells or erythrocytes. The blood transfusion from the wrong blood group can be life-threatening.

The four major blood groups are: A, B, AB and O. The blood grouping system was discovered in the year 1901 by Karl Landsteiner- an Austrian biologist and immunologist.

Also Read: Blood

Blood group testing

The main purpose of conducting this experiment is to understand the basic concept of the ABO blood group system and to know our blood group and type.

Materials Required

• Blood sample
• Alcohol Swabs
• Clean glass slide
• Sterile cotton balls
• Biohazard disposal container
• Monoclonal Antibodies (Anti-A, B, and D)

All these equipment will be readily available in a blood test tool kit.

• Take a clean glass slide and draw three circles on it.
• Unpack the Monoclonal Antibodies (MAB) kit. In the first circle add Anti-A, to the second circle add Anti-B and to the third circle add Anti-D with the help of a dropper.
• Keep the slide aside safely without disturbing.
• Now wipe the ring finger with the alcohol swabs and rub gently near the fingertip, where the blood sample will be collected.
• Prick the ring fingertip with the lancet and wipe off the first drop of the blood.
• As blood starts oozing out, allow it to fall on the three circles of the glass slide by gently pressing the fingertip.
• Apply pressure on the site where it was pricked and to stop blood flow. Use the cotton ball if required.
• Mix the blood sample gently with the help of a toothpick and wait for a minute to observe the result.

Here is the chart which predicts the different types of blood groups along with its Rh factor.

Precautions

Discard the alcohol swabs, lancet, cotton balls and toothpick after their use. Drop all the materials, including the glass slide into the biohazard disposal container after observing the result.

As mentioned above, there are four major blood groups and eight different blood types, collectively called the ABO Blood Group System. The groups are based on the presence or absence of  two specific antigens and antibodies– A and B:

• Group A- Antigen A and Antibody B.
• Group B- Antigen B and Antibody A.
• Group AB- Antigen A and B both and no Antibodies
• Group O- No Antigens and both A and B Antibodies.

Other than this, there is a third kind of antigen called the Rh factor. Based on the presence or absence of this antigen (Rh factor), the four blood groups are classified into eight different blood types:

• A positive – Presence of Rh+
• A negative- Presence of Rh-
• B positive- Presence of Rh+
• B negative- Presence of Rh-
• AB positive- Presence of Rh+
• AB negative- Presence of Rh-
• O positive- Presence of Rh+
• O negative- Presence of Rh-

Important Questions on  Blood Group Test:

• What is a blood group test?

A blood group test is a simple test used to determine the blood group of an individual. It is also called ABO typing.

• How many types of blood groups are there?

There are four major blood groups and are called the ABO Blood Group System. The types of blood groups are:

• Blood Group A
• Blood Group B
• Blood Group AB
• Blood Group O.

Which Blood group is called the universal donor and universal recipient?

Type O blood group is called the universal donor, as it can be donated to recipients of any blood type. This is because type O blood group neither have A or B on the surface of the red blood cells.

Type AB blood group is called the universal recipient and can receive blood from the donors of any blood type. This is because type AB blood group have both A and B  antigens on the surface of the red blood cells.

• What determines blood type?

The presence of antigen on the surface of the blood cell and the antibodies in the blood plasma can determine the blood groups or blood type of an individual. Moreover, these blood types or groups are inherited from our parents. Each parent passes one allele for blood type to their child. Therefore a child could have the same blood type as one of their parents.

• What’s the rarest blood type?

As per the records, AB-negative is considered to be the rarest blood type. It is difficult to say which blood type is the rarest in the world and it depends on the frequency in requirement of certain blood types, which varies widely in different parts of the world.

• Why is it important to know blood types?

The ABO blood types are important as they are essential during the blood transfusions and to avoid further complications.

Stay tuned with BYJU’S to learn more about the blood groups types, tests and other related topics at  BYJU’S Biology

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Blood Grouping PDF

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14 January 2020

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Blood Grouping Activity Sheet

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Blood Grouping and Rh Typing: Introduction, Principle, Procedure, Result- Interpretation, and Keynotes

Introduction.

Table of Contents

Blood Grouping and Rh Typing are used to determine the blood group and Rh factor of a sample of blood. ABO blood grouping and Rh typing are the most common blood group systems. There are nearly 300 blood group systems so far discovered. Other common remaining systems are MNS, Lutheran, Kell, Duffy, etc. Karl Landsteiner discovered ABO (A, B and O) in 1900. Two years later, Decastello and Sturli added AB and finally ABO system gets completed. In 1940, Karl Landsteiner and Winner discovered Rh factor in red blood cells (RBCs) of the Rhesus monkey (Macca rhesus).

For his great contribution to blood banking and transfusion medicine, he was awarded the Nobel Prize in Physiology or Medicine in 1930. He is also known as the Father of Blood Grouping and Immunochemistry. Methods of ABO Blood Grouping and Rh Typing are of two types i.e. slide and tube. But here we are concerned with the slide method of ABO blood grouping and Rh typing as it is rapid, simple, and easy to proceed.

ABO blood grouping and Rh typing are based on the principle of agglutination. The normal red blood cells (RBCs) possessing antigens will clump in the presence of corresponding antibodies.

Test Requirements

• Antisera A,B and Rhesus (Rh)
• Clean and grease-free glass slides
• Clean sticks for mixing (optional since slide can be used for this purpose)
• EDTA blood or clotted blood/ skin puncture using lancet at spot test
• Micropipette and tips/dropper-optional in case of skin puncture
• Physiological saline (optional)
• Label the two different clean slides with the unique ID of the patient.
• Pipette 50/50 µL (one-one drop) of the blood and deliver it to the first slide in two corners and a drop in the remaining slide at the center.
• Put 1 drop of A, B, and Rh antisera onto each slide respectively, and mix it thoroughly with the help of the clean sticks or using a clean glass slide.
• Tilt the glass slide back and forth for up to 2 minutes.
• Look for the agglutination in each slide. 
• In the case of Rh, if agglutination is seen it is reported as Rh positive and if no agglutination is seen, it is reported as Rh negative as shown table of result-interpretation.

Result Interpretation

Applications of ABO Blood Grouping and Rh Typing

It is applicable for the following streams-

• Prior to blood transfusion and cross-match.
• In the case of hemolytic disease of the newborn (HDN)
• Determination of paternity in medico-legal cases
• Relationship of blood groups, and susceptibility to a variety of diseases e.g. blood group A is more prone to gastric carcinoma whereas blood group O is more prone to systemic lupus erythematosus (SLE).
• In the case of doubtful agglutination reactions, a further microscopic examination should be done to confirm the result.
• Each negative Rh result should be confirmed by the Du Test.
• For clarification of the agglutination, add a drop of normal saline/physiological saline. Tilt the glass slide back and forth and observe for agglutination/clumping.
• To omit prozone and post zone phenomenon, use 10% suspension of RBCs in place of using whole blood.

Further Reading

• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4732084/
• https://en.wikipedia.org/wiki/ABO_blood_group_system
• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3497328/
• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5741336/
• https://nios.ac.in/media/documents/dmlt/hbbt/Lesson-07.pdf
• https://www.labpedia.net/blood-banking-part-1-blood-groups-abo-and-rh-system-blood-grouping-procedures/
• https://rfums-bigtree.s3.amazonaws.com/files/resources/abo.pdf
• https://www.jstor.org/stable/40793065?seq=1#metadata_info_tab_contents
• https://www.cambridge.org/core/journals/amg-acta-geneticae-medicae-et-gemellologiae-twin-research/article/karl-landsteiner-father-of-blood-grouping-and-immunochemistry/CB635080E85F850EC0694CFA169BE2B5

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Blood Group Test Procedure

Shares2Facebook2Tweet0Pin0LinkedIn0Email0 There are four main blood groups (types of blood) – A, B, AB and…

There are four main blood groups (types of blood) – A, B, AB and O. Your blood group is determined by the genes you inherit from your parents.

Each group can be either RhD positive or RhD negative, which means in total there are eight main blood groups.

Also Known As:   Blood Group ,  Rh Factor , ABO Grouping , Blood Typing , ABO Group

Blood is made up of red blood cells, white blood cells and platelets in a liquid called plasma. Your blood group is identified by antibodies and antigens in the blood.

Antibodies are proteins found in plasma. They are part of your body’s natural defenses. They recognize foreign substances, such as germs, and alert their immune system, which destroys them.

Antigens are protein molecules that are found on the surface of red blood cells.

ABO grouping can be performed in two ways Forward grouping/Cell grouping : Red blood cells are tested for A and B antigens using known anti-A and anti-B sera. Reverse grouping/Serum grouping : Serum is tested for anti-A and anti-B anibodies using known A and B red cells.

The ABO system

There are four main blood groups defined by the ABO system:

• blood group A –  has A antigens on the red blood cells with anti-B antibodies in the plasma
• blood group B   – has B antigens with anti-A antibodies in the plasma
• blood group O   – has no antigens, but both anti-A and anti-B antibodies in the plasma
• blood group AB   – has both A and B antigens, but no antibodies

Blood group O is the most common blood group. Almost half of the population (48%) has blood group O. 

The Rh system

Red blood cells sometimes have another antigen, a protein known as the RhD antigen. If this is present, your blood group is RhD positive. If it’s absent, your blood group is RhD negative.

This means you can be one of eight blood groups:

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

About 85% of the population is Rh positive (36% of the population has O+, the most common type).

Sample Required

• This can be done on whole blood or even on clotted blood.
• The sample can be stored at 4 °C. and stable for 5 days.
• Some time week subgroup may result in mistyping where Coombs test may be helpful.

Indications

• ABO blood grouping and Rh typing are done.
• Blood grouping is done for the donor and the recipient (Crossmatch).
• Blood grouping is done in the expected mother and newborn.

Test Procedure:

Blood grouping reagents

The blood Grouping reagents enables rapid identification of ABO blood group and Rh factor depending upon the antigen present on the surface of red blood cells.

Blood Grouping reagent

Blood grouping Procedure

• Inform the patient or individual about the procedure to be carry out
• Dangle the hand down to increase the flow of blood in the fingers.
• Clean the fingertip to be pierced with spirit or 70% alcohol (usually ring or middle finger) and gently massage the finger to increase blood flow
• With the help of the sterile lancet or pricker, pierce the fingertip and place one drop of blood in each of the four cavities
• Now add one drop of each antiserum into each cavity respectively

ABO and Rhesus Blood Grouping Tiles

• Mix each blood drop with the antiserum using a fresh  mixing stick  or applicator stick
• Now you can observe agglutination in the form of fine red granules within 30 seconds. Anti RhD takes slightly longer time to agglutinate compared to Anti A and Anti B.  

Blood Grouping Result and Interpretation

• ✓: Agglutination
• X : No agglutination  

Blood grouping result interpretation

• If  agglutination is observed when individual’s  blood is mixed with  Anti A reagent , then the individual is said to have a  blood group   “A” .  
• If agglutination is observed when individual’s  blood is mixed with  Anti B reagent , then the individual is said to have a blood group  “B”
• If agglutination is observed when individual’s  blood is mixed with  Anti A and Anti B reagent , then the individual is said to have a  blood group  “AB”
• If no agglutination is observed when individual’s blood is mixed with  Anti A and Anti B reagent , then the individual is said to have a blood group  “O”  
• If agglutination is observed when individual’s blood is mixed with  Anti RhD reagent , then the individual is said to have a  “+ve” Rh factor .
• If no agglutination is observed when individual’s blood is mixed with  Anti RhD reagent , then the individual is said to a have  “-ve” Rh factor.

Common setback during Blood grouping

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ABO Blood Group System

ABO blood grouping is based on the principle of an agglutination reaction . It is the popular method for blood group identification to determine the presence and absence of cellular antigens and their relative antibodies in the blood. In blood typing, the detection of antigen in the donor’s RBCs is called forward typing. In contrast, the  detection of antibodies in the donor’s plasma or serum is known as reverse typing .

According to the ABO-blood group system, A , B , AB and O are the four phenotypes. Type A and B are the two antigens associated with the RBC membrane, while anti-A and anti-B are the two antibodies naturally present in the blood plasma. The blood typing or the blood group identification is performed by the blood-test kit that contains anti-A, anti-B and anti-D antisera.

The antigens of the ABO blood group system are glycolipid in nature, and the antibodies are predominantly of IgM type. In this context, we will study the principle, method and result interpretation of the ABO blood typing. Besides, we will also discuss some of the discoveries, facts and the overview of the ABO blood group system.

Content: ABO Blood Group System

Interpretation of result, abo compatibility, meaning of abo blood group system.

The ABO blood group system has type A, B, AB and O phenotypes and it is used to identify the type of surface antigens and antibodies present in the donor’s blood. If agglutination occurs in the RBCs, then the corresponding antibody must be absent in the blood plasma. The antigens if absent on the RBCs membrane, then the corresponding antibodies must be present in the blood plasma. Individuals above 3-6 month have naturally occurring antibodies which arise without any antigenic stimulation from the maternal placenta. These antibodies belong to the IgM class. ABO blood group is also present in some other animals like gorillas, chimpanzees etc.

History of ABO Blood Grouping

According to the ABO blood group system, there are four blood groups, namely A , B , AB and O .

Method of ABO Blood Group System

• First, scrub the middle finger with cotton saturated with 70% of alcohol .
• Then, prick the middle finger by sterilized needle or lancet.
• After that, place three drops on a clean glass slide.
• Then after this, add antisera in a sequence of anti- A in a first drop, anti- B in a second drop and anti- D to the third drop, respectively.
• Mix the blood with the antisera separately by using a sterilized toothpick.
• Allow the slide to stand for 2-3 minutes and then note down the results based on clump formation or agglutination reaction.

• If the agglutination occurs in the RBCs, to which anti-A is added, then the blood group is ‘ A ’.
• When agglutination occurs in the RBCs, to which anti-B is added, then the blood group is ‘ B ’.
• If the agglutination occurs in the RBCs, to which both anti-A and B are added, then the blood group is ‘ AB ’.
• When there is no agglutination occurs in the RBCs, then the blood group is ‘ O ’.

In addition, there is also another antiserum that is anti-D , which determines the positive and negative blood type .

• If the agglutination occurs in the RBCs to which anti-D is added, then the blood type is positive (+) and if no agglutination occurs in the RBCs mixed anti-D, then the blood type is negative (-).

Theory of blood transfusion states that before transfusion, the ABO compatibility of blood type must be checked, as any carelessness can affect the immune system.

• A person with blood group A can receive blood from the person with blood types A and O.
• The person with blood group B can receive blood from the person with blood types B and O.
• A person with blood group AB can receive blood from the person with all blood types A, B, AB and O and called as Universal recipient .
• The person with blood group O can receive blood from only the person with blood type O.

• A person with blood group A can donate blood to the person with blood types A and AB.
• The person with blood group B can donate blood to the person with blood types B and AB.
• A person with blood group AB can donate blood to only the person with blood type AB.
• A person with blood group O can donate blood to the person with all blood types A, B, AB and O and hence called Universal donor .

Therefore, the ABO blood group system is one of the popular technique to classify human blood. ABO blood group system majorly classifies the blood into four types, i.e. A, B, AB and O. Agglutination reaction determines the ABO blood type, which is determined by the clump formation in the blood. ABO blood group system was accepted by the National Research Council and popularly known as Landsteiner classification .  Karl Landsteiner won the Nobel prize in the year 1930 for his contribution in this.

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Practical Experiment | Microbiology - Blood Grouping | 12th Microbiology : Practical Experiment Manual

Chapter: 12th microbiology : practical experiment manual.

Blood Grouping

To determine the blood group of the blood sample by the slide agglutination test.

Theory and Principle:

Blood grouping is an essential requirement before blood is transfused from one person to another.It is also useful in settling paternity disputes and medicolegal problems.

Red blood cells contain blood group antigens. Antibodies to the blood group antigens are present in the blood plasma.The antigens are generally determined and are responsible for blood types. When RBCs of a person are mixed with corresponding antiserum, agglutination occurs due to antigen-antibody reactions.

Materials Required

• Blood sample ( anticoagulated)

• Sterile cotton

• Sterile lancet

• Clean dry grease free slides or white tile

• Toothpicks

• Marker pen

• Commercially available Anti A sera, Anti B sera and Anti D sera

1. Prick the finger under aseptic conditions

2. Place a drop of blood on the slide on each side marked as A, B and D.

3. Add a drop of antiserum A , B and D on A, B and D side respectively.

4. Mix with toothpick using separate toothpicks for each mixture.

5. Wait for 2 mins and observe for clumping reaction if any confirm it by observing under microscope.

6. Interpret the results and report.

Interpretation

If agglutination on A side the blood group is A

If agglutination seen on B side the blood group is B

If Agglutination on both A and B side the blood group is AB

If No agglutination on A and B side the blood group is O

If agglutination is seen on D side the blood group is Rh(D) positive

If No agglutination on D side the blood group is Rh(D) negative.

Diagram: ( any one depending on the results)

Observation:

(will vary with the type of blood group an example is given below) Agglutination is seen on A, B and D side

The blood group of the blood sample was determined by slide agglutination test and was found to be AB Rh positive.

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