HIV INVITRO LAB STUDIES
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.