University:
Swansea University
Course:
PM134 | Microbiology
Academic year:

2023
Views:

51

Pages:

2
Author:

Sanjeev KG

EXPERIMENT NO: 15- DATE 6/12/22 DUCHTERLONY DOUBLE DIFFUCION AIN Te Hiedy the reaction patteer of an onligen with a tet of antibodies by Duchtertory Double disunion method PROCEDURE I fixedpace 10ml of 1% agrease (as given in important instructions). Copt the solution 55-60°c and pour 5ml/place on to geease tree glass plates placed on horizontal surface. Allow the gel to set for 30 min. 3. Place the glass plate on the template pasvicled. 4. Punch wells with the help of the gel puncher corresponding to the markings on the template torming of sugged wellb. Lese gentle suction to avoid 5. Add 10ml each of the to the welle as shown. antiserum of the corresponding antigens 6. keep the glass plate in a moist chamber overnight ar 37°c २ After incubation. observe for opaque precipitin lines. between the antiger Ef antisteum wells ABC OBLERVATION It was obscwed that Arc (Identity Precipelin Patterns) ene antisecum A showed Antiseeum B showed pattans) 4 alpur (Partial Odentity Precipiter Anosusum ( showed a intersect (Nan Identity-precipitin Pattern). REZULT A hub deneity Cie, a continuous line) - tation at theil junition farming. serologic identity of the presence of antigers when Antiselum A was added. Line of precipiー an are represent a common epitope in Pactint identity (ie, a continerous line with a spur at one end) in antigers The 2 antigens share a common epitope when antisicum B was added. Nos identity Cie, the 2 lines cross completely) - A crossed line pattern indicates that the El share added demonstrates no con mon 2 separate reactions af compared antigens. epitopes are Unrelated when antisceum ( urs DISCUSSION Ouchterlony immunodiffusion assay, developed by Sve dish physician Orjas Ouchterlony is used the ja the detection of antigens Ef antibodies & determinatio & homologies between antigens diffusion • Ouchtalony double is frequently used for comparing different antiger prepacation several diff antigens dies of the antiserum will be produced. It the antiger solution contains enai can react wels the antibo- then multiple lines of precipitatio The method is called 'double refelling' to the falt that in Procedwe allaved to migate towards antiger I antibody, all each other of precipitation is formed wheel meet in a gel af a line 2 reactants. This precipitation reaction is highly specific.