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Practical work #8. Definition of the blood group by ABO system with the help of coliclones
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Practical work #8. Definition of the blood group by ABO system with the help of coliclones

For the definition of the blood group in any system the same principle is used: providing conditions for erythrocytes agglutination in the medium of standard isohemagglutinating serums or coliclones, that have high titre of antibodies to the examined antigens of erythrocytes.

Objectives: To define the blood group by ABO system.

Requirements for work: white plate, pipettes, glass, pencil for glass, examined blood, closed tubes with solutions of coliclones anti-A and anti-B, isotonic solution of sodium chloride.

divide dry white plate on 4 sectors with glass-pencil.

make notes “anti-A”, ”anti-B”.

with the help of pipettes put drops (0,1 ml) of coliclones anti-A and anti-B in the accordant sector. Make a string using the same pipette (diameter of the string should not be less than 1,5 – 2 cm).

put one drop of the examined blood on the glass.

using the angles of another glass put blood (0,01 ml) in the drops of coliclones. Angles of glass should be different for different coliclones.

mix blood with the drop of coliclone with the angle of glass. Correlation between blood and coliclone should be 1:10 (mixed drop has pink color).

observe the reaction on the plate during 2,5 minutes.

Results and control

Agglutination does not occur with coliclone anti-A and with coliclone anti-B. So, examined erythrocytes do not have antigens A and B, and blood belongs to group I (0, ??).

Agglutination occurred only with coliclone anti-A. So, examined erythrocytes have only antigen A and blood belongs to group II (A, ?).

Agglutination occurred only with colicone anti-B. So, examined erythrocytes have only antigen B and blood belongs to group III (B, ?).

Agglutination of erythrocytes is observed in both drops of coliclones. So, examined erythrocytes have both antigens A and B and blood belongs to group IV (AB).

It should be mentioned that all processes that occur after 2,5 minutes after mixing will not be connected with specific agglutination, which is examined and those can have other reasons. False agglutination can occur when erythrocytes will gather in monetary column. This agglutination can be easily discerned from the real one if added 1-2 drops of isotonic solution of sodium chloride to 1 drop of blood. False agglutination will disappear in this case.

Recommendation for writing down the results:

Write down the results of agglutination reaction with coliclones anti-A and anti-B in the following chart:

           coliclone

blood

Anti-A

Anti-B

І (0, ??)

 

 

ІІ (А, ?)

 

 

ІІІ (В, ?)

 

 

ІV (АВ)

 

 

 Answer the following questions in conclusion:

What means presence/absence of agglutination of the examined blood with coliclone anti-A?

What means presence/absence of agglutination of the examined blood with coliclone anti-B?



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