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CHECK-LIST Criteria of statement assessment of hemagglutination-inhibition reaction (HIR) for typing influenza virus

CHECK-LIST Criteria of assessment statement of hemagglutination reaction

¹ Criteria of Assessment
State the purpose of the hemagglutination reaction assay. Completely describe the steps of the hemagglutination reaction assay and name the components of the reaction
Hemagglutination reaction is run on microtiter plates
In 6 microwells add virus dilutions from 1:2 till 1:64. The 7th well is for erythrocyte control
4. Add 0,5 ml of 1% suspension of chicken erythrocytes to each microwell. Plate is left at room temperature for 30 minutes
5. In negative control microwells, note a precipitation of erythrocytes in the form of "button". A positive reaction will have a deposit in the form of "umbrella" in the microwell. Comment on the received result. Explain the reaction mechanism

CHECK-LIST Criteria of statement assessment of hemagglutination inhibition reaction (HIR) for detection of antibody

¹ Criteria of Assessment
State the purpose of hemagglutination inhibition assay for detection of antibodies. Completely describe the steps of the hemagglutination inhibition assay for detection of antibodies and name the components of the reaction
Patient serum is diluted from 1:10 to 1:320 with physiological saline in microwells of a microtiter plate
In all microwells, add 0,25 ml virus diagnosticum (antigen) of corresponding disease. Then add on 0,5 ml 1% suspension of chicken erythrocytes. Prepare three wells as controls: 1. KS – serum + erythrocytes 2. KV – virus + erythrocytes 3. KE – physiological saline + erythrocytes Incubate 45 minutes.
Positive reactions form a deposit in the form of "button" to the corresponding antigen. In virus control KV – a deposit in the form of "umbrella", in erythrocytes control KV– in the form of "button", in serum control KS– in the form of "button".
Comment on the results. Explain the reaction mechanism.

CHECK-LIST Criteria of statement assessment of indirect hemagglutination reaction (IHR)

¹ Criteria of Assessment
State the purpose of the IHR assay. Completely describe the steps of the IHR assay for detection of antibodies and name the components of the reaction
Add 0,5 ml of normal saline to each microwell of a microtiter plate, then add 0,5 ml of test serum diluted 1:25 to the first well in each row. Make two-fold dilutions by transferring 0,5 ml from one well to the next in the row, making final dilutions ranging from 1:50 to 1:800
Add 0,1 ml of erythrocyte diagnosticum antigen to each well Reaction is accompanied by 2 controls: 1. KA – antigen control– physiological saline and antigen 2. KS – control of serum – physiological saline and serum. Incubate microplate at 37oC for 2 hours
Positive reactions and control KS form a deposit in the form of "button" to the corresponding antigen. In negative control KA – a deposit in the form of "umbrella”
Comment the results. Explain the reaction mechanism.

CHECK-LIST Criteria of statement assessment of hemagglutination-inhibition reaction (HIR) for typing influenza virus



¹ Criteria of Assessment
State the purpose of the HIR assay for definition of influenza virus typing. Completely describe the steps of the HIR assay for determination the type of influenza virus and name the components of reaction
Add 0,25 ml of a virus content material to each of 5 wells in a microtiter plate. Add 0,25 ml of each standard type antiserums against influenza virus ( A0, A1, A2, B, C) per well
Add 0,5 ml of 1% of chicken erythrocytes to each well and incubate at room temperature for 1 hour
The influenza type will be the well in which the deposit in the form of "button" was formed.
Comment the results. Explain the reaction mechanism.

Date: 2015-12-18; view: 614


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