Methods
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Micro Haemagglutination Inhibition Test
Procedure
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Dispense 25 µl of PBS / normal saline into each well of the plates.
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Shake each serum sample and dispense 25µl into the first well and the last well (serum control) of a row of a micro well plate.
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Use a micro pipette to make two-fold serial dilutions along the row until the second last well from the end. The last well is the serum control. Do not dilute this well.
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Add 25 µl of the 4HA dilution of antigen to each well excluding the control wells
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Gently tap the sides of the micro well plates to mix the reagents. Cover plates with a lid.
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Allow for 30 minutes at room temperature.
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Add 25 µl of 1 percent washed red blood cells to each well including the control wells in the last column.
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Gently tap the sides of the micro well plates to mix the reagents. Cover the plates with a lid.
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Allow standing at room temperature for 45 minutes.
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RBC Control: Add 25 ul of saline to well to which add 25 ul of 1% chicken RBC
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Check Virus Titration
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Add 25 ul of saline into four wells in a row.
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Add 25 ul of the 4HA virus suspension to the first well and make serial two fold dilution.
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Discard 25 ul of the suspension from the fourth well.
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Add 25 ul of 1% chicken RBC to all the four wells.
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Leave for 20 minutes.
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The first two wells will show haemagglutination , if the viral suspension prepared is having 4HA units of virus in every 25 ul .
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Read the settling patterns for each serum sample.
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Read the control serum well first then read the patterns in the other wells.
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Last modified: Sunday, 25 September 2011, 10:13 AM