WO2017018939A1 - Procédés et trousse pour la différenciation d'anticorps vih-1 et vih-2 - Google Patents

Procédés et trousse pour la différenciation d'anticorps vih-1 et vih-2 Download PDF

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Publication number
WO2017018939A1
WO2017018939A1 PCT/SG2016/050352 SG2016050352W WO2017018939A1 WO 2017018939 A1 WO2017018939 A1 WO 2017018939A1 SG 2016050352 W SG2016050352 W SG 2016050352W WO 2017018939 A1 WO2017018939 A1 WO 2017018939A1
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Prior art keywords
hiv
solid support
immunodeficiency virus
human immunodeficiency
antigens
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PCT/SG2016/050352
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English (en)
Inventor
Weiping Hu
Yun Ying Tan
Yuik Chen Tracy PANG
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Mp Biomedicals Asia Pacific Pte Ltd
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Publication of WO2017018939A1 publication Critical patent/WO2017018939A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • G01N33/56988—HIV or HTLV
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
    • G01N2333/08—RNA viruses
    • G01N2333/15—Retroviridae, e.g. bovine leukaemia virus, feline leukaemia virus, feline leukaemia virus, human T-cell leukaemia-lymphoma virus
    • G01N2333/155—Lentiviridae, e.g. visna-maedi virus, equine infectious virus, FIV, SIV
    • G01N2333/16—HIV-1, HIV-2
    • G01N2333/162—HIV-1, HIV-2 env, e.g. gp160, gp110/120, gp41, V3, peptid T, DC4-Binding site
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2469/00—Immunoassays for the detection of microorganisms
    • G01N2469/20—Detection of antibodies in sample from host which are directed against antigens from microorganisms

Definitions

  • the present invention relates generally to the fields of virology and immunology. In particular, it relates to the field of diagnostic assays. More particularly, the present invention provides a Point-Of-Care test (POCT) that can detect the serology markers of human immunodeficiency virus in a sample.
  • POCT Point-Of-Care test
  • HIVs Human Immunodeficiency Viruses
  • AIDS acquired immunodeficiency syndrome
  • HIV was first discovered in two separate research groups independently in 1983. Two types of HIV have been identified and characterized: HiV-1 and HIV-2. Both types are transmitted by sexual contact, through contaminated blood, body fluids and from mother to child during pregnancy or breastfeeding. HIV-1 is the major cause of HIV infection and AIDS in the world. HIV-2 is less common and less infective. Studies of the geographic distribution of HIV infections reveal that HIV-1 is the predominant type worldwide, HIV-2 is largely concentrated in West Africa.
  • HIV infections elicit immune responses and induce HIV specific antibodies in human.
  • the serological methods such as Enzyme-linked Immunosorbent Assay (ELISA), Colloidal Gold Immunochromatographic Rapid Test, and Western Blot (WB) are widely used in the laboratory diagnosis of HIV-1 and HIV-2 infection.
  • ELISA Enzyme-linked Immunosorbent Assay
  • WB Western Blot
  • the present invention is a qualitative immunochromatographic assay for the rapid in vitro detection and differentiation of antibodies to HIV-1 and HIV-2 in human serum, plasma, finger pricked whole blood or whole blood with anti-coagulants. It is intended for professional use as a rapid screening test for the presence of antibodies to HIV in patients and aid in the diagnosis of HIV-l/HIV-2 infection. The positive results should be further confirmed by more specific supplemental tests such as Western Blots.
  • a method for detecting a human immunodeficiency virus infection in a subject comprising (a) providing human immunodeficiency virus antigens; (b) contacting the human immunodeficiency virus antigens with a biological sample obtained from the subject, the biological sample suspected of containing human antibodies against the human immunodeficiency virus antigens; (c) detecting the presence of a complex that forms between the human immunodeficiency virus antigen and the human antibody, wherein the method simultaneously and differentially detects the presence of antibodies in the sample to HIV-1 and HIV-2 antigens, and the presence of the complex is indicative of a HIV-1 and/or HIV-2 infection in the subject.
  • the method further comprising (a) providing a solid support having a first end and a second end; (b) immobilising the human immunodeficiency virus antigens on the solid support, the antigens are immobilised separately and spaced apart from each other along the solid support; (c) immobilising a molecule on the second end of the solid support, the molecule capable of binding to the human antibody in the complex, the molecule further comprising at least one detectable labelled moiety; (c) loading the sample on the first end of the solid support, the sample contacting the antigens as it travels along the solid support in a first direction along the solid support and towards the second end; and (d) loading a buffer on the second end of the solid support adjacent the molecule such that the buffer solution solubilise the molecule as it travels along the solid support in a second direction opposite the first direction, wherein the presence of the complex formed on the solid support is detected by the labelled moiety of the molecule.
  • the method further provides for an absorbent pad that is placed adjacent to or abuts the first end of the solid support, the absorbent pad and the solid support are separated by a separator, wherein removing the separator allows contact between the absorbent pad and the solid support to enhance the flow of the buffer along the solid support in the second direction, for example to increase the flow rate of the buffer.
  • the human immunodeficiency virus antigen may be a structural protein of human immunodeficiency virus particles and fragments thereof, or any glycoproteins, lipids and carbohydrates derived from the human immunodeficiency virus. More preferably, the method comprises using all the human immunodeficiency virus antigens gpl20, gp41, gp36 and p24.
  • the human antibody is IgG.
  • Any such binding method may be used that is known to a person skilled in immunoassay techniques.
  • the molecule is a human immunodeficiency virus antigen.
  • the molecule is an anti-human antibody.
  • the anti-human antibody is a goat anti-human IgG.
  • the detectable labelled moiety is a gold conjugate.
  • the biological sample is selected from the group comprising of whole blood, plasma, and serum.
  • a kit for detecting a human immunodeficiency virus infection in a human subject comprising a solid support with human immunodeficiency virus antigens, wherein the human immunodeficiency virus antigens are gpl20, gp41, gp36 and p24 and the antigens are immobilised on the solid support separately and spaced apart from each other.
  • the solid support further comprising a molecule immobilised on it, the molecule capable of binding to a complex formed with the surface-bound human immunodeficiency virus antigens, the molecule further comprising at least one detectably labelled moiety.
  • a sample obtained from the subject is loaded on a first end of the solid support, and a buffer for solubilising the molecule is loaded on a second end of the solid support opposite the first end.
  • the kit further comprises an absorbent pad adjacent the first end of the solid support, the absorbent pad and the solid support are separated by a separator, wherein removing the separator allows contact between the absorbent pad and the solid support to enhance the flow of the buffer along the solid support.
  • the molecule is an anti-human antibody.
  • the anti-human antibody may be a goat anti-human IgG.
  • the molecule may also be a human immunodeficiency virus antigen.
  • the detectable labelled moiety is a gold conjugate.
  • the solid support includes a nitrocellulose membrane.
  • the present method and kit allows for a quick and easy detection of a human immunodeficiency virus infection in a sample taken from a patient.
  • the method and kit can provide results that can differentiate whether a patient is HIV-1 and/or HIV-2 positive.
  • the present method and kit allows for increased sensitivity compared to conventional HIV screening tests and have proven early seroconversion detection.
  • Figure 1 is a schematic drawing of a kit according to an embodiment of the present invention.
  • Figure 2 is a schematic drawing of using a kit according to embodiment of the present invention.
  • Figure 3 is a schematic drawing showing possible results on a kit according to an embodiment of the present invention.
  • Figure 4 is a schematic drawing showing a cross section of the kit according to an embodiment of the present invention.
  • Figures 5, 6 and 7 are charts showing the profiles of visible test lines.
  • the present invention provides for a kit 5 for detecting a human immunodeficiency virus infection.
  • the present kit is a Point-of-Care screening test kit that is capable for the simultaneous and differential detection of HIV infections (HIV 1 and/or HIV-2) with multiple markers.
  • HIV infections HIV 1 and/or HIV-2
  • the kit 5 includes a casing having an inlet Well A 20 for receiving a biological sample obtained from a subject, another inlet Well B 25 for receiving a chaser buffer (which will be described more later), and a reaction chamber 30.
  • the reaction chamber 30 may be the portion of the kit 5 which includes a window allowing a user to see the results obtained when the assay method is carried out on the kit 5.
  • the inlet 20 may also include a sample chamber for receiving the sample.
  • the inlets 20, 25 and reaction chamber 30 are all in fluid communication with each other.
  • Inlet 20 is opposite inlet 25.
  • the kit 5 also includes any solid support structure 10 having human immunodeficiency virus antigens immobilised on it.
  • the solid support 10 may be made of any suitable material, for example an absorbent material, that spans the inlets 20, 25 and includes the reaction chamber 30 where the human immunodeficiency virus antigens are immobilised.
  • the front of the casing of the kit 5 adjacent the window may include markings showing the locations of the various test lines (and hence, human immunodeficiency virus antigens that are immobilised on the solid support structure 10).
  • the antigens are immobilised on the solid support in a "striped” and linear fashion as shown in Figure 1.
  • These antigens may be immobilised on the nitrocellulose membrane using a Dispensing system, followed by a drying step.
  • the "striped" membrane is subjected to blocking step with blocking solution containing Casein, Triton, PVP and 5X stabil coat for 1.5 min, followed by a drying step.
  • the antigens may be immobilised by any suitable methods known to the skilled person
  • the human immunodeficiency virus antigens gpl20, gp41, gp36 and p24 are immobilised on the solid support structure 10. This is shown on the markings adjacent the window of the kit 5. These 4 human immunodeficiency virus antigens (or test lines) are arranged separately and spaced apart in a linear fashion such that they line up along the length of the window of the kit 5.
  • An indicator line 40 (in an embodiment, the indicator line may be blue) is shown on the solid support structure 10 to indicate when the sample has travelled across the immobilised gp36, p24 ; gp41, gpl20 and protein A along the length of the solid support structure 10.
  • a control line 45 may be included.
  • having all four human immunodeficiency virus antigens derived from HIV-1 and HIV-2 virus on a single strip of solid support structure 10 allows for an assay to be carried out to determine the serology of the HIV detected in a sample.
  • the kit is an indirect solid-phase immunochromatographic assay that is based on a reverse flow technology disclosed in US Patent No. 6,316,205 (contents of which are incorporated herein) and may be used for the simultaneous and differential detection of antibodies to HIV-1 and HIV-2 in human serum, plasma, finger pricked whole blood or whole blood with anti-coagulants.
  • the support structure may be a nitrocellulose membrane.
  • the antigens may be obtained commercially and purified by chromatography.
  • the kit may further include any molecule that is capable of binding to a complex that may be formed with the human immunodeficiency virus antigen immobilised on the membrane and a human antibody against the human immunodeficiency virus antigen.
  • the molecule may be an anti-human antibody.
  • the anti-human antibody is goat anti-human IgG.
  • the goat anti-human IgG further conjugated to colloidal gold for detection of the bound complex.
  • the molecule may be a human immunodeficiency virus, or part of it.
  • Figure 4 shows the kit of the present invention in greater detail, in particular it shows a cross section of it.
  • the components shown are separated from each other. In the actual kit, these components are in contact with each other.
  • the solid support 10 is part of a strip which includes a sample pad 50, a test strip 55, and a gold conjugate pad 60.
  • the sample pad 50 receives the sample from the inlet 20.
  • the solid support 10, particular the test strip 55 includes the immobilised antigens 35 (test markers).
  • the test strip 55 may be made from a material that is different to that of the sample pad 50 and gold conjugate pad 60.
  • the test strip 55 is made of a nitrocellulose membrane. This is the part of the strip that is shown in the window 30.
  • the gold conjugate pad 60 has the molecule that is capable of binding to the antibody-antigen complex. This molecule has a gold conjugate for detection purposes.
  • the gold conjugate pad 60 receives the buffer from the inlet 25.
  • the sample pad 50 is at a first end of the strip while the gold conjugate pad 60 is at a second end of the strip. First and second ends are opposite each other.
  • an absorbent pad 70 At the first end of the strip, i.e. the sample pad 50, is an absorbent pad 70.
  • the absorbent pad 70 is placed beneath the sample pad 50.
  • the absorbent pad 70 and sample pad 50 are separated by a separator 75, i.e. the "pull tab" as shown in the Figures.
  • a sample is received in inlet 20 and gets absorbed by the sample pad.
  • the sample then travels along the strip in the direction A to the test strip 55 by capillary action where it comes into contact with the antigens that are immobilised on the test strip 55. If there are any HIV antibodies in the sample, anti-HIV antibodies will bind to the immobilised antigens and antibody-antigen complexes will be formed on test strip 55 at the relevant marker positions as indicated in 35.
  • a buffer is then received in inlet 25 and re-solubilise the molecule on the gold conjugate pad 60. When the separator 75 is removed by pulling the tab, the sample pad 50 and absorbent pad 70 comes into contact.
  • a biological test sample may be obtained from a subject (for example, a patient) that may be suspected of being infected with the human immunodeficiency virus (HIV).
  • the biological sample taken from such patients may contain human antibodies against a human immunodeficiency virus antigen.
  • the antibodies in the biological test sample serum, plasma, finger pricked whole blood or whole blood with anti-coagulants
  • Whole blood can be collected in anti-coagulant containing tubes and used as outlined in the assay procedure immediately or stored at 2°C to 8°C for not more than 72 hours before use.
  • ACD Acid Citrate Dextrose
  • CPD Citrate-phosphate-dextrose
  • EDTA Ethylenediaminetetraacetic acid
  • K-Oxalate Potassium Oxalate
  • Lithium Heparin Li Hep
  • Na Citrate Na Citrate
  • Serum / plasma samples should be stored at 2°C to 8°C if the test is to be run within 7 days of collection or frozen at -20°C or colder if the test is to be delayed for more than 7 days. Clear, non-hemolyzed samples are preferred. Lipemic, icteric or contaminated (particulate or bacterial) samples should be filtered (0.45 ⁇ ) or centrifuged before testing.
  • the assay method for detecting a human immunodeficiency virus infection in a subject may be carried out on the kit 5.
  • the method includes providing human immunodeficiency virus antigens; contacting the human immunodeficiency virus antigens with a biological sample obtained from the subject that is suspected of containing human antibodies against the human immunodeficiency virus antigens; and detecting the presence of a complex that forms between the human immunodeficiency virus antigen and the human antibody.
  • the present method simultaneously and differentially detects the presence of antibodies in the sample to HlV-1 and HIV-2 antigens.
  • the presence of the complex is indicative of a HIV-l and/or HIV-2 infection in the subject.
  • the antigens target human IgG.
  • the human immunodeficiency virus antigens are immobilised on a solid support structure 10.
  • a sample for example, serum, plasma or whole blood
  • chase buffer may be added to the Well A 20 right after the sample addition.
  • the sample chamber which may be part of or is in fluid communication with the Well A 20, is also in fluid communication with the reaction chamber 30 of the kit. As such, the sample will start travelling or wicking up the solid support structure 10 in a first direction A as shown in Figure 2.
  • the buffer may contain sodium chloride, Sodium di hydrogen phosphate monohydrate, sodium azide, EDTA Na2.2H20, Triton X-100 and 10% SDS.
  • the anti-human IgG gold conjugate in buffer chamber Well B 25 will be resolubilised by the chase buffer and travel along a buffer support 10 in a second direction (B) opposite the first direction (A).
  • the anti-human IgG gold conjugate comprises any molecule that is capable of binding to the human antibody in the complex. Such a molecule further comprising at least one detectable labelled moiety.
  • the molecule may be an anti-human antibody.
  • the molecule may be a human immunodeficiency virus antigen or an anti-human antibody.
  • the anti-human antibody may be a goat anti-human IgG.
  • the moiety for detection may be a gold conjugate. Other types of detection systems such as biotin/avidin may be used.
  • the sample pad and absorbent pad are separated by a separator (pull tab), and removing the separator allows the contact of sample pad and absorbent pad, which facilitate the flow downwards in direction B.
  • the presence of the complex is detected by detecting the moiety.
  • the separator may be a material that is impermeable.
  • the separator is the tab 15 shown in Figure 1.
  • the bound antibody-antigen complexes are subsequently detected by goat anti-human IgG gold conjugate carried by a chase buffer that flows downward (direction B) giving a pink- purplish colour.
  • the control line 45 may contain Protein A captures human IgG from the patient's sample, subsequently binds with the anti-human IgG goid conjugate.
  • the appearance of control line 40 serves to validate the proper addition of human specimens and migration of sample and chase buffer, as well as the re-solubilised anti-human IgG gold conjugate.
  • Protein A or recombinant Protein A may be obtained from Staphylococcus aureus Cowan I that is expressed in Bacillus, lmg of Recombinant Protein A binds to at least lOmg of human IgG.
  • the gold conjugate is re-solubilised when 3 drops added to the oval well (Well B).
  • the gold conjugate is sprayed and dry on the gold conjugate pad 60. Once chase buffers added, the buffer will dissolve the gold conjugate and the gold conjugate in liquid format will flow downwards (direction B).
  • chase buffer After pull-tab pulled, 1 drop of chase buffer was used to make contact between sample pad and absorbent pad to facilitate the flow downwards.
  • the drop of chase buffer connects the sample pad portion of the solid support to the absorbent pad.
  • the kit 5 is allowed to warm to room temperature (25°C ⁇ 3°C) before running the assay for optimal binding. For best results, the assay is conducted at room temperature (25°C ⁇ 3°C).
  • the assay method is diagrammatic represented in Figure 2.
  • the "HIV” Pull Tab 15 is pulled until resistance is felt. Another 1 drop of chase buffer is added into the square well [A]. The results are read at 20 minutes but before 25 minutes.
  • "Pull Tab” is to separate two stage of flow. Before pull-tab is pulled, it functions as separator to avoid contact between absorbent pad and sample pad, and thus the specimens flow upwards (direction A) by capillary reaction; after pull-tab 75 is pulled, the absorbent pad contacts with sample pad and facilitate the flow of gold conjugate downwards (i.e. the direction where the anti-human IgG gold conjugate flows towards well A - Direction B).
  • a test is HIV-1 positive if it meets any of the following criteria:
  • Control Line (C) appears with visible gpl20 test line (with or without visible gp41 and/or p24 test lines);
  • Control Line (C) appears with visible gp41 test line (with or without visible gpl20 and/or p24 test lines);
  • Control Line (C) appears with visible gpl20 & gp41 test lines (with or without visible p24 test line).
  • a test is HIV-2 positive if Control Line (C) appears with visible gp36 test line (with or without visible p24 test line)
  • a test is HIV-1 and HIV-2 positive if it meets any of the following criteria:
  • Control Line (C) appears with visible gpl20 & gp36 test lines (with or without visible gp41 and/or p24 test lines);
  • Control Line (C) appears with visible gp41 & gp36 test lines (with or without visible gpl20 and/or p24 test lines);
  • Control Line (C) appears with visible gpl20 & gp41 & gp36 test lines (with or without visible p24 test line).
  • a test is HIV positive (untypable) if Control Line (C) appears with visible p24 test line only. A test is negative if Control line (C) appears and Test line(s) are not visible.
  • a test is invalid if Control line (C) is absent.
  • the assay should be repeated using a new device. Performance of the present HIV Rapid Test
  • a total of 801 anti-HIV positive specimens and 2057 anti-HIV negative specimens collected from various countries were tested on the present HIV Rapid Test at MP Biomedicals Asia Pacific Pte Ltd- R&D Laboratory or ICDDR, Bangladesh (Clinical evaluation). As shown in Table above, the present HIV Rapid Test demonstrated 100 % sensitivity (95% CI : 99.54% to 100.00%) and 99.12% specificity (95% CI: 98.62% - 99.48%). The >99% accuracy based on the tested population (n 2858) indicating the good performance of the present HIV Rapid Test. The positive predictive value and negative predictive value for the present HIV Rapid Test were found to be 97.80% and 100% respectively.
  • the present HIV Rapid Test detected all 801 specimens as positive, demonstrating 100% sensitivity (95 % CI: 99.54% - 100%).
  • a total of 2057 specimens comprised of seven categories (healthy donors, cross-reactive, interference substances, hospitalized, clinical, anti E-coli positive, pregnancy specimens) were tested on the present HIV Rapid Test at MP Biomedicals-R&D laboratory or International Centre for Diarrhoeal Disease Research (ICDDR), Bangladesh (clinical site).
  • HIV antigens gp41, p24 and gp36
  • the potential cross-reactivity with anti-E.co// ' positive human specimens were examined.
  • 20 anti-E.co// ' positive human specimens were tested on the present HIV Rapid Test at ICDDR, Bangladesh.
  • the present HIV Rapid Test did not show cross-reactivity with all 20 anti-E.co// positive samples, resulting in 100% specificity (20/20).
  • HIV Rapid Test demonstrated 99.12% total diagnostic specificity using 2057 specimens, with 95% CI as 98.62% to 99.48%. Differentiation of HIV-l and HIV-2
  • the present HIV Rapid Test detects and differentiates between anti HIV-l and anti HIV-2 antibodies, by four separate test lines (gpl20, gp41, p24 and gp36) on the test devices.
  • Highly purified recombinant antigens gpl20 and gp41 representing HIV-l, gp36 representing HIV-2 and p24 reacts with both HIV-l and HIV-2.
  • anti HIV-l positive specimens Of 506 anti HIV-l positive specimens, 1 specimen was detected as positive (untypable), 493 specimens were detected as positive for HIV-l antibody and 12 specimens were detected as positive for HIV-l and HIV-2 antibody on the present HIV Rapid Test. Of 108 anti-HIV-2 specimens, 100 specimens were correctly detected as positive for HIV-2 and 8 specimens were found to be reactive with both HIV-l test lines and HIV-2 test lines on the present HIV Rapid Test.
  • the accuracy of the differentiation of the present HIV Rapid Test was determined. Out of 506 HIV-l specimens, 493 specimens were considered accurately detected for HIV-l serotype (with reactive gp41 and/or gpl20 test lines). On the other hand, out of 108 HIV-2 specimens, 100 specimens showed reactive HIV-2 (gp36) test line and considered correctly differentiated for HIV-2 serotype.
  • the 24 HIV-1 & HIV-2 positive specimens determined by MPD HIV Blot 2.2 and MPD HIV-2 Blot 1.2 were showing concordant results with reactive HIV-1 and HIV-2 markers on the present HIV Rapid Test. Thus, the 24 samples were included in both HIV-1 and HIV-2 differentiation accuracy analysis respectively.
  • the overall differentiation accuracy by the present HIV Rapid Test was found to be 97.55% (517/530) for HIV-1 and 93.94% (124/132) for HIV-2.
  • anti-HIV 1 specimens comprised of various subtypes from Group M and 4 anti-HIV 1 specimen was from Group O; the remaining 4 specimen was from HIV-2.
  • various subtypes from Group M tested on MULTISURE HIV Rapid Test included subtype A, B, C, D, E, F, G, H, J, K, BC, C/E as well as circulating recombinant forms (CRF) derived from recombination between viruses of different subtypes (i.e.CRF01_AE, CRF02_AG,CRF02_AB, E/F, G/CRF02, H/Al, K/CRF09 & CRF01/CRF15).
  • the present HIV Rapid Test detected all available HIV-1 Group M subtypes, the HIV Group O subtype and HIV-2 as positive.
  • the comparative performance on seroconversion sensitivity of the present HIV Rapid Test was evaluated against various other types of Rapid Test kits that are available.
  • the results of commercials Rapid Test diagnostic kits were extracted from COA of seroconversion panels from Zeptometrix Corporation and SeraCare Life Sciences. Referring to Table 2, in 5 seroconversion panels, the present HIV Rapid Test and Inverness Determine HIV 1/2 Ag/Ab Combo kit detected all 5 seroconversion panels as positive.
  • the average days of detection since first bleed of the present HIV Rapid Test are comparable with Inverness Determine HIV 1/2 Ag/Ab Combo kit, which were 26.8 days and 27.6 days respectively (For the calculation for the average days of detection, in case where an assay is negative over all seroconversion follow up bleeds, two additional days are assigned on the last bleed for the calculation).
  • the overall sensitivity of the present HIV Rapid Test 38.89%, was better than Inverness Determine HIV 1/2 Ag/Ab Combo kit (30.56%).
  • the average days of detection since first bleed for the present HIV Rapid Test, Biorad Multispot HIV Rapid Test, Oraquick Advance HIV 1/2 Rapid Ab and Trinity BioTech Uni Gold Recombinant HIV-1 were determined based on 10 seroconversion panels.
  • the present HIV Rapid Test showed the earliest average days of detection since first bleed (48.5 days), followed by Biorad Multispot HIV Rapid Test (51.3 days), Trinity BioTech Uni Gold Recombinant HIV-1 (55.1 days) and Oraquick Advance HIV 1/2 Rapid Ab (55.4 days).
  • the present HIV Rapid Test demonstrated the highest reactivity (41.18%) as compared to other rapid diagnostic kits (28.57% - 35.29%).

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Abstract

La présente invention concerne un test de point d'intervention (POCT) qui peut détecter les marqueurs sérologiques du virus immunodéficitaire humain (VIH) dans un échantillon. L'invention concerne un procédé destiné à détecter une infection au VIH chez un sujet comprenant (a) la fourniture d'antigènes du VIH ; (b) la mise en contact des antigènes du virus immunodéficitaire humain avec un échantillon biologique obtenu auprès du sujet, l'échantillon biologique étant soupçonné de contenir des anticorps humains vis-à-vis des antigènes du VIH ; (c) la détection de la présence d'un complexe qui se forme entre les antigènes du VIH et l'anticorps humain, le procédé détectant simultanément et différentiellement la présence d'anticorps dans l'échantillon vis-à-vis des antigènes VIH-1 et VIH-2, et la présence du complexe est indicative d'une infection au VIH-1 et/ou au VIH-2 chez le sujet. L'invention concerne également une trousse pour effectuer le dosage. Le procédé et la trousse revendiqués sont illustrés par l'utilisation de gp120, gp41, gp36 et p24 en tant qu'antigènes de VIH-1 et de VIH-2.
PCT/SG2016/050352 2015-07-27 2016-07-27 Procédés et trousse pour la différenciation d'anticorps vih-1 et vih-2 WO2017018939A1 (fr)

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CN109406781A (zh) * 2018-12-27 2019-03-01 正元盛邦(天津)生物科技有限公司 一种可降低传染风险的hiv（1+2）抗体检测卡及其适配试纸条
CN109406781B (zh) * 2018-12-27 2023-04-11 正元盛邦(天津)生物科技有限公司 一种可降低传染风险的hiv（1+2）抗体检测卡及其适配试纸条
CN111273004A (zh) * 2020-03-09 2020-06-12 北京华晟源医疗科技有限公司 一种基于胶体金法检测尿液中HIV(l+2)抗体试剂条及其制备方法
CN111273004B (zh) * 2020-03-09 2024-01-19 北京华晟源医疗科技有限公司 一种基于胶体金法检测尿液中HIV(l+2)抗体试剂条及其制备方法

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