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HIV-1 p24 antigen acridine ester chemiluminescence immune analyse detecting method

A technology of acridinium ester chemistry and luminescent immunity, which is applied in the field of clinical blood detection and analysis, can solve the problems of no acridine, etc., and achieve the effect of being conducive to automatic operation, reducing operation steps and high sensitivity

Inactive Publication Date: 2008-10-08
BEIJING UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] However, there is no report on the use of acridinium ester chemiluminescence immunoassay detection method for HIV-1p24 antigen detection

Method used

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  • HIV-1 p24 antigen acridine ester chemiluminescence immune analyse detecting method
  • HIV-1 p24 antigen acridine ester chemiluminescence immune analyse detecting method
  • HIV-1 p24 antigen acridine ester chemiluminescence immune analyse detecting method

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Embodiment

[0063] 1) Dilute the anti-p24 antibody for coating to 6 μg / ml with coating buffer, add 100 μl to each well of a white 96-well plate, and block at 4°C for 16 hours;

[0064] 2) Shake off the liquid in the wells, add 300 μl of washing buffer to each well for washing, wash for 3 minutes each time, and wash 3 times in total;

[0065] 3) Add 300 μl of blocking buffer to each well, and shake at a constant temperature of 37°C for 1 hour;

[0066] 4) Shake off the liquid in the wells, add 300 μl of washing buffer to each well to wash, wash for 3 minutes each time, wash 3 times in total, dry and store at 4°C;

[0067] 5) Add five positive control samples with concentrations of 80, 40, 20, 10, and 5 pg / ml into the wells, and add 100 μl to each well for 2 replicate wells at each concentration, and add negative control samples to 4 wells respectively. Add 100 μl to each well, add 40 samples (N1-N20, P1-P10 and L1-L10) of HIV-1p24 Antigen National Reference Substance to each well, add 100...

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Abstract

HIV-1 p24 antigen acridine ester chemiluminescence immunity analyzing testing method pertains to the clinical blood testing analysis method technique field. The prior HIV-1 p24 antigen testing method has problems of low sensitivity, rigmarole operations and the like. The invention uses acridine ester series compounds to mark the HIV-1 p24 antibody, adopts a double antibody / antigen sandwich method to execute immunity combination between the p24 antigen-antibody; uses excitant hydrogen peroxide, nitric acid, Triton X-100, and sodium hydrate to excite acridine ester series compounds to execute chemiluminescence reaction; and tests the number of photon to execute a qualitative and / or quantitative analysis. The invention has good correlativity with the general ELISA testing method, which has a correlation coefficient of 0.951 at the instance of P no more than 0.01; has high sensitivity, wherein, the testing limit is 0.5pg / ml; has wide detection range in 5-6 magnitude order; and the invention also has advantages of short reaction time, easier operation and the like.

Description

technical field [0001] The invention belongs to the technical field of clinical blood detection and analysis methods, in particular to a method for clinical and laboratory detection of HIV-1p24 antigen. The method has high sensitivity, wide detection range, and simple operation, and has important applications in the diagnosis of neonatal HIV-1 infection, monitoring of HIV-1 infected patients' disease progression, drug resistance monitoring, and evaluation of antiviral treatment effects. Background technique [0002] After HIV-1 (Human immunodeficiency virus) virus infects the human body, some marker substances of HIV infection will be produced in host cells and viruses (Niel T.Constantine, Holly Zink.HIV testing technologies after two decades of evolution.Indian J Med Res 121.2005 :519-524), through these markers, we can carry out virus detection, monitor the replication of the virus in the body, understand the development of the disease after infection, and grasp the status...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/543G01N21/76G01N33/545
Inventor 冯娟马雪梅钟儒刚曾毅
Owner BEIJING UNIV OF TECH
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