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In invitro lab studies, this drug was put for Drug Sensitivity Test and Anti-retroviral Activity Test in the laboratory adhering to the standard protocol and the highest inhibition rate of 98% was recorded against the causative agent, HIV. The detail procedures undertaken in an invitro lab study are mentioned as under: –

Procedure of the invitro lab study
  • Collect 20 ml Blood in an EDTA containing vial
  • Mix it properly and keep at room temperature
  • Overlay this blood sample on 2 ml Histopaque solution in 25-ml sterile centrifuge tube. Centrifuge the tube at 1500 rpm for 20 min
  • Two layers will be formed after centrifugation
  • Separate out the upper layer of Histopaque with the help of sterile pipette and collect the lymphocyte ring in 2 ml sterile eppendorf tube.
  • Microfuge it for 1 min at the bottom of the eppendorf tube lymphocytes sediment will be observed.
  • Resuspend the sediment in 2-ml fresh RPMI-C Media.
  • Prepare the serial dilutions of the drug of our interest or to be tested in the range of 1ug/ml – 500 ug/ml
  • Add 500 ul of the lymphocyte culture with the help of sterile pipette / tips and add the appropriate dilution of the drug to the respective culture well along with control without addition of any drug.
  • Incubate it in CO2 incubator at 37 degree Celsius (5% CO2)
  • After incubation do the P-24 ELISA test for Control and for Test
  • Interpret the results on the basis of difference in between the Control O.D and Test O.D and calculate the % Inhibition of the viral growth in Test in comparison with Control.
Example : O.D of the Control always ranges from 0.2-0.4 while the O.D of Test ranges in between 0.06-0.1, which indicates 98% inhibition of the virus multiplication.

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