A liquid-chip HIV-antigen antibody combined detection kit and a detecting method

An antigen-antibody, combined detection technology, applied in the field of immunoassay, can solve the problem of high false positive rate, and achieve the effect of ensuring blood transfusion safety, early detection, and improving detection sensitivity

Inactive Publication Date: 2016-04-27
GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to overcome the problems of the high false positive rate of HIV third-generation reagents and the limitations of WB detection, the present invention proposes a combined detection kit and detection method for HIV antigen and antibody by liquid phase chip method

Method used

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  • A liquid-chip HIV-antigen antibody combined detection kit and a detecting method
  • A liquid-chip HIV-antigen antibody combined detection kit and a detecting method
  • A liquid-chip HIV-antigen antibody combined detection kit and a detecting method

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Experimental program
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Effect test

Embodiment 1

[0219]

[0220]105 specimens from Tianjin Blood Center were analyzed, luminexxMAP detected 8 positive specimens in 19 specimens with indeterminate WB but positive RNA, and 3 negative specimens were detected in luminexxMAP in 18 specimens with indeterminate WB but negative RNA Specimens, indicating that luminexxMAP can reduce the occurrence of indeterminate specimens to a certain extent. Among the 3 WB-negative but RNA-positive samples, the detection result of luminexxMAPHIV-1 antibody in 1 case was indeterminate, which avoided the missed detection of WB to a certain extent, indicating that the specificity of luminexxMAP was higher than that of WB. The sensitivity of LuminexxMAP detection was 82.7% (72 / 87) higher than that of WB 74.7% (65 / 87).

Embodiment 2

[0222]

[0223] Analysis of the test results of 33 cases of RNA-positive gay men and women showed that 13 cases were positive for HIV-1 antibody by WB, 18 cases were indeterminate samples for HIV-1 antibody, and 2 cases were negative for HIV-1 antibody; 19 cases were positive for luminexxMAP. Uncertain specimens were 14 cases. By comparison, it was found that the detection sensitivity of luminexxMAP was 57.8% (19 / 22) higher than that of WB (39.4%).

Embodiment 3

[0225]

[0226] HIV immunoblotting test judgment criteria have always been based on antibody detection: the formation of at least two Env bands (gp41, gp120 / gp160) or one Env band plus p24 band is judged as positive, and the band type does not meet the positive judgment criteria. The result is judged as positive Suspicious, no HIV banding results are judged as negative, which can identify false positive results of the primary screening test. image 3 The results show that the detection of luminexxMAP antigen can suggest to a certain extent that the specimen may be before or during the positive conversion of the antibody, but the HIV-1p24 antigen is transiently increased in the infected person, so two cases of HIV-1, M1126 and M1127, also appeared Antibody positive, HIV-1p24 antigen negative.

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Abstract

The invention discloses a liquid-chip HIV-antigen antibody combined detection kit and a detecting method and belongs to the field of immunoassay. The kit comprises microspheres respectively covered with HIV gp41, p17, p24, p31, p66 and gp120 antigens, a biotinylated anti-human secondary antibody, an SA-PE solution, microspheres covered with a p24 monoclonal antibody, and a monoclonal antibody labeled with fluorescein and matching the p24 monoclonal antibody. Combined detection for six HIV antibodies and a P24 antigen can be performed simultaneously through a small amount of a sample. Compared with an immunoblotting method, the kit and the detecting method are high in sensitivity and specificity. Through the kit and the detecting method, the HIV antibodies can be detected timely in the earlier period of infection, thus achieving early detection and early intervention of HIV infection, reducing HIV transmission and ensuring safety of blood transfusion. The kit and the detecting method have a wide application prospect.

Description

technical field [0001] The invention belongs to the field of immunoassays, in particular to a combined detection kit and detection method for HIV antigen and antibody by liquid phase chip method. Background technique [0002] HIV infection detection includes P24 antigen, HIV antibody, HIV viral load, CD4 cell count and other methods. Currently, the most commonly used method for clinical HIV blood screening is enzyme-linked immunosorbent assay (ELISA). HIVELISA reagents have been developed to the 4th generation so far. At present, the mainstream reagents and methods for HIV detection in the world and at home are the third-generation reagents, the double-antigen sandwich method detects antibodies in the specimen, and the enzyme marker is a specific HIV antigen, which can detect different HIV antibody subtypes HIV-1 / 2 / OIgG / M / A, its detection sensitivity and specificity are higher than those of the first and second generation reagents, which improves the reactivity of group O...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/577G01N33/543
CPCG01N33/56988G01N33/54313G01N33/577G01N2333/161G01N2333/162
Inventor 袁玉华孔伟伟李艳
Owner GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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