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Avicenna EIA , HIV1+2 Ag/Ab ( Fourth generation)

Avicenna EIA-HIV1+2 Ag/Ab test system is a qualitative, enzyme immunoassay for the detection of circulating antibodies to Human Immunodeficiency Virus type 1(HIV1) and /or Human Immunodeficiency Virus Type 2(HIV2) in human serum or plasma and detection of p24 core antigen of HIV virus Type 1.

INTRODUCTION

Human Immunodeficiency Virus Type (HIV1) has been isolated from patients with AIDS and AIDS –related complex (ARC). HIV-1 was thought to be the only causative agent of these syndromes until 1986, when a second type of Human Immunodeficiency Virus (HIV2) was isolated and also reported to cause AIDS. Since the initial discovery more than few hundred cases of HIV-2 infection have been documented worldwide. Both viruses have the same morphology and lymphotropism, and the modes of transmission appear to be identical. In addition, HIV1 and HIV2 genomes exhibit about 60% homology in conserved genes such as gag and pol. Serologic studies have also shown that the core proteins of HIV–1 and HIV-2 display frequent cross-reactivity whereas the envelope proteins are more type-specific.
Despite this immunologic cross-reactivity, detection of antibodies to HIV-2 with any of the licensed HIV-1 enzyme immunoassay is highly variable. This device was developed to detect antibodies to HIV-1 and HIV-2 for blood screening and diagnostic purposes.
The human monoclonal used in the new generation HIV screening test have been shown to be insensitive to high titters of rheumatoid factors and not reacting to serum with HAMA.

Besides these properties, as the natural host of HIV is human not mice; anti-p24 HIV antibodies efficiently bind and recognize naturally occurring epitopes. When solid-phase surfaces are sensitized with such antibodies; the circulating p24 proteins are better captured and subsequently detected by a complementary anti-p24 labelled antibody.
Furthermore, the adaptive immunity to HIV generates sharply focused antibodies by molecular fitness and maturation. In contrast to antibodies derived from mice immunizations; human antibodies to p24 are able to recognize different structures of circulant p24 particles, and not only the immunizing p24 protein.

Any specimen that reacts in an initial test with the HIV-1 and HIV-2 must be retested in duplicate with Avicenna anti HIV1+2 –EIA. Repeated reactive specimens may contain antibodies to either HIV-1 or HIV-2. Therefore, additional, more specific or supplemental tests such as immunoblot, immunofluorescence and /or PCR must be performed to verify presence of antibodies to HIV.
This is an enzyme-linked immunosorbent assay using recombinant antigen and synthetic proteins corresponding to a highly antigenic segment of HIV-1/HIV-2 envelope proteins and monoclonal antibodies to detect HIV-1 antigen.


PRINCIPLE OF THE ASSAY

Multiple Epitopes of HIV1+2 proteins and antibodies to p24 are bound to the microplate wells. When antibodies to HIV-1/HIV-2 and/or p24 antigen are present in the test sample, they react with synthetic and recombinant proteins and/or p24 capture antibodies and attach to the solid-phase. Non-reactive antibodies are removed by the washing solution.

During conjugate 1 incubation, biotinylated detector antibody/detector antigens will bind to antibodies/antigen if present. After removing conjugate 1, peroxidase labelled streptavidin is added which binds to the biotinylated antigens/antibodies (conjugate 2). After a second wash step, substrate is added which is converted to a blue-coloured product.

Positive sample generates a dark blue color. No color or very pale blue color indicates a negative reaction. After stopping the reaction by adding stop solution, the color changes from blue to yellow. The intensity of the reaction is photometrically quantitated.


TECHNICAL SPECIFICATIONS

Intended Purpose: Enzyme Immunoassay test designed for qualitative determination of Antibodies to HIV-1, HIV-2 and p24 antigen in serum/plasma Principle: Sandwich antigen principle for antibody detection and sandwich antibody principle for antigen detection.


REAGENTS
Materials provided with the kits :

  • Coated Plate: Two micro plates, each with 96 wells coated with HIV-1/HIV-2 synthetic and recombinant proteins and anti p-24 monoclonal. It is sealed in a foil pouch. Code: 0305/PL.

  • Sample Diluent: One bottle of 30 ml chemically defined solution containing salts, detergent and antimicrobial agent. Code: 0305/SD.

  • Positive Control: One vial (3.0 ml) of heat inactivated human serum containing anti- HIV-1/HIV-2 –antibodies carried by Elisa method and confirmed by Western Blot. (Non- reactive for Hepatitis B surface antigen and antibodies to HCV carried out by Elisa method and confirmed by Western Blot.). With addition of preservative solution and Rodamine.

  • Negative Control: One vial (3.0ml) of heat inactivated human serum non- reactive for Hepatitis B surface antigen, antibodies to HIV nor HIV-1 p24 antigen and HCV carried out by Elisa with addition of preservative solution.

  • Conjugate 1(100X): One vial with 0.5 ml concentrated C1 (biotinylated antigens/antibodies). Code: 0305/C1.
  • Conjugate Diluent 1: 1x50 ml bottle containing blue color diluents for concentrated C1. Code: 0305/CD1.
  • Conjugate 2 (100x): One vial with 0.5 ml concentrated C2 (peroxidise labelled streptavidin). Code: 0305/C2 .
  • Conjugate Diluent 2: 1x50 ml bottle containing orange color diluent for concentrated C2. Code: 0305/CD2.
  • Chromogen (TMB) (100x): One bottle (0.5ml) of concentrated TMB. Code: 0305/TMB.

  • Substrate buffer: 1x50 ml bottle containing transparent peroxide containing buffer for dilution of concentrated TMB. Code: 0305/SB.

  • Washing Solution (10X): Three bottles (2X50+1X25ml) chemically defined solution containing NaCl, NaOH, Tween 20 and 5.7 % acetic acid.

  • Instruction manual: one copy .

Avicenna HCV -EIA
Anti HCV- EIA-Avicenna kit is a qualitative, enzyme immunoassay for the detection of antibodies to hepatitis C virus (anti-HCV) in human serum or plasma.

INTRODUCTION

Hepatitis C virus (HCV), which was formerly described as the parenterally transmitted from non-A, non-B hepatitis (NANBH), causes chronic disease in 60% of the cases. HCV can be transmitted by intravenous drug abuse, sexual, and household contact. Hepatitis C virus is a single stranded RNA virus that is related to the flavivirus family. Nucleic acid sequences of HCV provided the basis for the construction of recombinant and synthetic peptides representing putative hepatitis C virus proteins. Anti-HCV screening of blood using synthetic or recombinant proteins identified apparently healthy blood donors infected with HCV. This is an enzyme immunosorbent assay (EIA) using recombinant and synthetic proteins derived from core and non-structural regions of HCV to detect the presence of anti-HCV in human serum or plasma samples.

The figure below shows the arrangement of the HCV genome and the amino acid sequences used in the anti-HCV- EIA-Avicenna


Core
(a.a. 2-60)

NS3
(a.a.1192-1457)

NS4
(a.a.1918-1940)

NS5
(a.a 2385-2404)

PRINCIPLE OF THE ASSAY

Multiple Epitops of HCV proteins are bound to the microplate wells. When antibodies to HCV are present in the test sample, they react with the synthetic and recombinant proteins attached to the solid-phase. Non-reactive antibodies are removed by the washing solution. The immune-complex is reacting with conjugate and visualized by subsequent reaction with TMB substrate. Positive sample generates a dark blue color. No color or very pale blue color indicates a negative reaction. After the reaction stops the color changes from blue to yellow. The intensity of the reaction is measured photometrically.

TECHNICAL SPECIFICATIONS

Intended Purpose

Enzyme Immunoassay test designed for qualitative detection of anti-HCV in serum or plasma

Production Principle

3rd Generation Method using direct binding principle.

Sensitivity

100% (with International Standard Reference Panels of WHO, VQC, BBI and Russian national panel)

Specificity

99.9%. No cross reactivity with Superoxide Dismutase (SOD), Flavivirus and yellow fever virus infection.

The assay fulfils the criteria for performance evaluation in the Common Technical Specifications (CTS) of the European Directive for In vitro Diagnostic Devices (IVD) 98/79/EC.


Specimen

Serum or Plasma

Capture Antigens

Contains Synthetic and Recombinant
HCV Core, NS3, NS4, and NS5 antigens

 

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