HIV ELISA 4th Gen

HIV ELISA 4th Gen

ProLISA HIV 4th Gen is an in-vitro enzyme immunoassay for the qualitative determination of antibodies to HIV-1/HIV-2 and p24 antigen in human serum or plasma. It is intended for screening of blood donors or other individuals at risk for HIV-1 or HIV-2 infection and for clinical diagnostic testing.

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Overview:

HIV 4.0 ELISA is an in vitro enzyme immunosorbent assay for qualitative determination of antibodies to HIV-1/HIV-2 and HIV-1 p24 antigen in human serum or plasma.

  • HIV type I and type II (HIV1+HIV2) are enveloped around single-stranded RNA positive virus.

  • HIV-1 has been isolated from patents with AIDS and AIDS-related complex and from healthy individuals with a high risk for developing AIDS.

  • HIV virus is transmitted through-

  • Sexual contact

  • Exposure to blood

  • Transmitted from an infected mother to fetus during parental period.


 

Principle:

“Double antibody and antigen sandwich immunoassay.”

  • Recombinant of HIV-­1(gp41) and HIV-­2 (gp36) antigens and anti-­p24 antibodies are coated onto microwells.

  • Biotinylated sample diluent and serum or plasma samples are added to these wells. The wells are washed to remove unbound components.

  • After washing wells to remove unbound enzyme, substrate is added. Colour will develop in wells.

  • No colour will develop in wells having no antigen-­antibody complex.

  • Colour is read on photometer at a wavelength of 450nm-650nm.



 

Procedure:

  1. Add 25µl of sample diluent in each well

  2. Cover the plate with aluminum foil and incubate for 60mins at 37°CAdd 75µl of control or specimen in separate wells

  3. Read results at 450/620nmAdd 100µl of stop solution to stop reaction

  4. Add 100µl of substrate solution in each well and incubate at RT for 30mins

  5. Wash each well 5 mes with 300µl of wash buffer and dry with absorbent paper

  6. Add 100µl of conjugate in each well and incubate at 37°C for 30 mins

  7. Wash each well 5 times with 300µl of wash buffer and dry with absorbent paper

  8. Add 100µl of substrate solution in each well and incubate at RT for 30mins

  9. Add 100µl of stop solution to stop reaction

  10. Read results at 450/620nm


 

  • Store all kit components at 28°C when not in use.

  • Sensitivity 100% , Specificity> = 98%