The aim of the given practical is the introduction to biological techniques like blood grouping. Biological techniques involving procedures like blood type grouping is mentioned which is performed with the help of ABO blood group testing methods as well as the determination of the RhD or the Rheumatoid or Rh factor in the individuals. The blood groups of the parents and daughters are given and their genetic information is deduced from the possible cobinantion of blood groups. From the results of the given experiment conducted on the possible ABO blood grouping, the possiple genotypes of the father and mother has also been determined in the upcoming paragraphs. Moreover, the possible combinantions of the genotyes between parents and their offsprings have been demonstrated though the representation of a family tree.
Background:
The term blood group is often characterized by the entire blood system which comprises the various blood groups. The specificity is often controlled by series of genes which can be determined as allelic or linked closely and are present on the similar chromosomes (Mitra, Mishra and Rath, 2015).ABO method is the most common procedure of blood typing. Blood typing is the kind of tests done which determine the kind of blood and often is fruitful in finding out the genetic relation with other people. The ABO blood typing method isuseful in finding the compatible blood type during blood transfusion experiments and for cases involving blood donation. This method is based on the types of antigens present in the blood cells and results aredrawn from the agglutination reactions(Mydr.com.au, 2019). According to Holocomb et al. (2015), blood transfusion methods have been successful in the improvement of patient survival suffering from trauma after translocation of thawed plasma from the reserved blood banks. Similarly,Barnett et al. (2014) suggests that ABO blood grouping and incompatibility in renal transplantation have been an effective strategy for enhancement of the population of the potential donors for the patients who lack a suitable compatible donor for transplantation. Such protocols involve the techniques of preventing the eventual return of the antibodies after transplantation along with removal of the antibodies in the immediate pre-transplant period. According to statistical analysis done on the blood typing of cancer patients, it has been found that they effective in the determination of the type of cancers and also can observe the correlation of the types of cancers with the determined blood types (Li et al. 2014).
RhD is another method of determination of blood type based on the Rhesus system. According to the experiment cells are termed as Rh+ if there is the presence of Rh antigen on the surface of the cells. The absence of such antigen is termed as Rh- or Rh negative. As far as the significance of the RhD blood typing is concerned, prediction of the RhD type of the fetus have been successful and its integration into clinical practice have led to the successful deliveries of babies where the blood group of their mothers are Rh –ve. Furthermore, the noninvasive prediction of the RhD type of the fetus can be applied as a guide for targeting the prenatal prophylaxis and avoiding unnecessary exposure to the antigen in pregnant women (Clausen 2014). Rhd factor and its preence also helps in the detection of risk development of diseases like diabetes mellitus. The investigation of blood groups are evident in the future clinical as well as epidemiological factors required. From the studies importance have been given to people with blood group O regarding the risk of diseas developments. Another significance of RhD methodhas been its implementation in transfusion medicine. This has reduced the cases of hemolytic transfusion reactions related t the transfusion of incompatible blood. In order to be clinically significant, all donors for blood transfusion have been tested for ABO and RhD methods for the eterminatio of their respective blood groups (Schoeman et al.2017).
Materials and methods:
For conducting blood typing experiments, 4 bottles containing the four antigens lie Anti-A, Anti-B, Anti AB, Anti-D are used. The samples from the parents are required where dilutions have to be done and 3% suspensions are used. For the samples of the daughter 3% red cell suspensions are required for the students named D1,D2,D3,D4 . Moreover, 3% suspension of A as well as red cells are used. Apart from it, Pasteur pipettes, test tube rack and markers are required.
Firstly, the four antibodies will be distributed into the tubes one drop in each tube. Care should be taken so that the drops reach the end of the tubes. Then 1 drop of the cell suspension is given into the tubes and shaken properly. This step is followed by the incubation in the benches for almost5-10 minutes and centrifugation of the tubes is done at 1000 rotations perminute or rpm for 20 seconds.
Secondly for serum grouping, 2 drops of the daughter or parent plasma is given and 1 drop of the reagent test cells (A,B cells) into the respective tubes. Itshould be checked that the serum as well as the cells are properly mixed together and then incubation is done at room temperature for 1 minutes. Centrifugation is done at 1000 rpm for 20 seconds. Re-suspension of the cells are done and each test of agglutination is assessed macroscopically.
Results:
Agglutination is doneon the basis of the agglutination scoring chart from a grade of 0 to 4 where a negative result is received for no agglutination. Then a grade of 1 to 4 is given on the basis of agglutination where small agglutinate form level 1 and +4 is an exemplification of very strong levels of agglutination and there is no such presence of free cells.
From the results it can be seen that there has been no such agglutination between the serums of parent P1. However therehas been no agglutination between the parent and daughter D2.Similarly,there has been no agglutination between the serum of P2 and the same results have been seen for the samples of the daughters named D3,D4. Strongest levels of agglutination have been seen in the samples of D1 and D2 for antibody A. and A,B both. However there has been no agglutination observed in the daughters D3,D4 for B antibody and A, B.
Discussion:
From the results, it can be seen that for parent A there has been the presence of both antigen resulting in strong agglutination reactions. Thus the blood group is AB+ as both the A and B antigens have been preset resulting in agglutination and the positive sign is due to the presence of the Rh factor. For patient B there has been no agglutination for Anti B antisera it shows the absence of antigen B. Thus, only antigen A is present as there has been strong agglutination with the antibody of A. Hence, the blood group of parent B would be Anegative due to the absence of the Rh factor. Since D4 or the fourth daughter has not shown agglutination with any of the antibodies, the blood group is O.
Part 2: Interpretations
Answer 1: The possible genotype for parent 1 is AB as both the A and B alleles are present. Since both the alleles are present there cannot be more than one available genotype on this blood group.
Answer 2:The possible genotypes for the 2nd parent is either AA or AO as the antigen A has been identified and they can have either one allele or both the alleles. According to the ABO system of blood grouping the available genotypes would be either homozygous, heterozygous. Among the homozygous groups the genotypes can be either homozygous dominant (AA in this case) and Homozygus recessive (ii in this case).
Answer 3:
Case 1:
AB AO
AA AO AB BO
Case 2:
AB AA
AA AA AB AB
Answer 4:If daughter 1 or D1 receives Rh positive blood, then there will be production of antibodies within the body where the Rh antigen is absent. An immunogenic reactiondevelops between the Rh positive and Rh negative blood known as transfusion. Thus a transfusion reaction will occur which can seriously affect her progeny in the future if she is pregnant and her babies have Rh +ve blood. When the blood of mother will reach the foetus, the antibodies against the Rh antigens will destroy them. A result of this transfusion reaction is anemia and most often resulting in a phenomenon known as erythroblastosis foetalis. Nowadays Rh negative mothers are given doses of Rh immunoglobulin or Rhogham is administered carefully for the preventing the destruction of antibodies.
Conclusion:
From the following paragrphs it can be
concluded that blood grouping plays an important role in the determination of
blood groups. The ABO and Rh systems are useful in the dtermination and provide
information based on the presence and absence of the A and B antigens. The
presence of antigen in the blood group reacts with the antisera which lead to
agglutination. The determination of the type of blood group present in parents
can give useful information of the possible blood groups of children. Moreover
genotypes can also be determined and other useful information regarding the
genetic similarity between parentsand children can be observed from the family
trees. Thus, the use of ABO systems and family trees helps in the genotypic
analysis and linkage between individuals.
References
Banch Clausen, F., 2014. Integration of noninvasive prenatal prediction of fetal blood group into clinical prenatal care. Prenatal diagnosis, 34(5), pp.409-415.
Barnett, A.N.R., Manook, M., Nagendran, M., Kenchayikoppad, S., Vaughan, R., Dorling, A., Hadjianastassiou, V.G. and Mamode, N., 2014. Tailored desensitization strategies in ABO blood group antibody incompatible renal transplantation. Transplant International, 27(2), pp.187-196.
Carson, J.L., Stanworth, S.J., Roubinian, N., Fergusson, D.A., Triulzi, D., Doree, C. and Hebert, P.C., 2016. Transfusion thresholds and other strategies for guiding allogeneic red blood cell transfusion. Cochrane database of systematic reviews, (10).
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Holcomb, J.B., Donathan, D.P., Cotton, B.A., del Junco, D.J., Brown, G., Wenckstern, T.V., Podbielski, J.M., Camp, E.A., Hobbs, R., Bai, Y. and Brito, M., 2015. Prehospital transfusion of plasma and red blood cells in trauma patients. Prehospital Emergency Care, 19(1), pp.1-9.
Klein, H.G. and Anstee, D.J., 2014. Mollison’s blood transfusion in clinical medicine. John Wiley & Sons.
Mydr.com.au. 2019. Blood typing – ABO blood groups and Rh types. Retrieved from https://www.mydr.com.au/tests-investigations/blood-typing
Rohde, J.M., Dimcheff, D.E., Blumberg, N., Saint, S., Langa, K.M., Kuhn, L., Hickner, A. and Rogers, M.A., 2014. Health care–associated infection after red blood cell transfusion: a systematic review and meta-analysis. Jama, 311(13), pp.1317-1326.
Schoeman, E.M., Lopez, G.H., McGowan, E.C., Millard, G.M., O’Brien, H., Roulis, E.V., Liew, Y.W., Martin, J.R., McGrath, K.A., Powley, T. and Flower, R.L., 2017. Evaluation of targeted exome sequencing for 28 protein‐based blood group systems, including the homologous gene systems, for blood group genotyping. Transfusion, 57(4), pp.1078-1088.
Wager, K. A., Lee, F. W., & Glaser, J. P. 2017. Health care information systems: a practical approach for health care management. John Wiley & Sons.
