Non-diagnosis-targeted HIV antibody immunity detecting method and kit

An immune detection method and immune detection technology, applied in the field of HIV antibody detection, can solve the problems of high test cost, high price, and many operation steps, and achieve the effect of easy automation, simple operation, and strong stability

Inactive Publication Date: 2016-10-12
THE THIRD XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Methods such as GPAT and dot immune colloidal gold rapid test are subjective, expensive, and difficult to store.
The disadvantage of CLIA is that the luminescent reaction of labeled catalytic enzymes or chemiluminescent molecules is unstable, which is intermittent and flashing, and it is prone to fission during the reaction process, resulting in unstable reaction results.
In addition, the binding phase and t

Method used

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  • Non-diagnosis-targeted HIV antibody immunity detecting method and kit
  • Non-diagnosis-targeted HIV antibody immunity detecting method and kit
  • Non-diagnosis-targeted HIV antibody immunity detecting method and kit

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Embodiment 1

[0052] The various solution formulas used in the embodiment are as follows:

[0053] Sodium phosphate-sodium chloride buffer solution (sodium phosphate-sodium chloride buffer solution, SPSC) (50mM Na 2 HPO 4 / 1.0M NaCl): 17.9g of NaCl 2 HPO 4 12H 2 O, dissolve 58.5g of NaCl with a little ultrapure water, adjust the pH to 7.5 with hydrochloric acid, and set the volume to 100mL.

[0054] 10mM phosphate buffer solution (phosphate buffer solution, PBS) (pH=7.4): 8g NaCl, 0.2g KCl, 1.44g Na 2 HPO 4 12H 2 O, 0.24g KH 2 PO 4 Dissolve it in 1L of ultrapure water, adjust the pH to 7.4 with hydrochloric acid, and store it at 4°C after autoclaving.

[0055] Preparation of Biotin-I: 3.3 nmol Biotin-I synthesized according to known sequence was dissolved in 33 μl ultrapure water to 100 μM, and diluted to 5 nM with SPSC buffer. Aliquot and store at -20°C for later use. Avoid repeated freezing and thawing before use.

[0056] Preparation of Biotin-H1 and H2: 2.1nmol H1 synthesized...

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Abstract

The invention relates to a non-diagnosis-targeted HIV antibody immunity detecting method and a kit. The method comprises the following steps: specifically capturing and detecting an HIV antibody in a sample by using an HIV antigen, then adding a biotinylated secondary antibody, and incubating to form an antigen-antibody-biotinylated secondary antibody compound; adding a biotinylation primer Biotin-I in the presence of streptavidin, incubating to combine the compound with the Biotin-I, and then adding biotinylated hairpin chain Biotin-H1 and hairpin chain H2 to carry out hybridization reaction; adding avidin marked horseradish peroxidase, catalyzing a substrate solution to develop color, and carrying out quantitative detection on the HIV antibody by determining absorbance. HCR is guided into ELISA, signals are amplified by the HCR, ultra-sensitive detection of the HIV antibody is realized, and lower limit of detection is as low as 9.5 pg/mL, and is higher than that of the traditional ELISA by about 2 orders of magnitude.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, in particular to a non-diagnostic HIV antibody detection method and kit. Background technique [0002] Human immunodeficiency virus (Human immunodeficiency virus, HIV) is the pathogen that causes acquired immunodeficiency syndrome (Acquired Immune Deficiency Syndrome, AIDS). HIV antibody is produced by the human immune response after the HIV virus enters the human body. This antibody is basically ineffective against the HIV virus, but it can reflect the HIV infection status to a certain extent and is a physiological indicator of the human body. [0003] Detecting HIV antibodies in blood is currently the most commonly used laboratory method for detecting HIV infection. Generally, it needs to go through two steps: first, do a preliminary screening test, and if it is positive, then do a confirmatory test. Only when the confirmation test is positive can the diagnosis of AIDS be made. Viral infe...

Claims

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Application Information

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IPC IPC(8): G01N33/569
CPCG01N33/56988G01N2333/16G01N2469/20
Inventor 聂新民桂嵘董彩霞黄蓉贺思涵
Owner THE THIRD XIANGYA HOSPITAL OF CENT SOUTH UNIV
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