Detection method for AMACR auto-antibody and its application in prostatic cancer diagnosis

A technology of autoantibodies and detection methods, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of low concentration and unsuitability for prostate cancer markers, and achieve the effects of high purity, low detection cost, and simple and easy operation

Inactive Publication Date: 2009-01-14
无锡纳生生物科技有限公司
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Problems solved by technology

However, the concentration of AMACR in the blood is

Method used

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[0038] 1. Preparation of reagents

[0039] (1) Coating buffer (0.05M, pH9.6): 1.59g Na 2 CO 3 , 2.93g NaHCO 3 , Add ultrapure water to dilute the volume to 1000mL;

[0040] (2) Phosphate buffer (PBS, 0.15M, pH7.4): 8.0g NaCl, 0.2gKCl, 2.9gNa 2 HPO 4 ·12H 2 O, 0.2g KH 2 PO 4 , Adjust the pH to 7.4 and add ultrapure water to dilute the volume to 1000mL;

[0041] (3) Phosphate-Tween buffer (PBST, 0.15M, pH7.4): 8.0g NaCl, 0.2gKCl, 2.9gNa 2 HPO 4 ·12H 2 O, 0.2g KH 2 PO 4 , 0.5ml Tween-20, adjust the pH to 7.4 and add ultrapure water to make the volume to 1000mL;

[0042](4) Blocking solution: 1.0g gelatin is dissolved in 100ml phosphate buffer (PBS, 0.15M, pH7.4);

[0043] (5) Substrate buffer (0.05M, pH5.6): 39.8g Na 2 HPO 4 ·12H 2 O, 6.3g citric acid, add ultrapure water to make the volume to 1000mL;

[0044] (6) Chromogenic solution: 10mL pH5.6 substrate buffer, 0.1mL 1% tetramethylbenzidine (10mg tetramethylbenzidine dissolved in 1mL dimethylsulfoxide) and 32μL 0.75% H 2 O 2 , Mix...

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Abstract

The invention discloses an AMACR auto-antibody test method and the application of the method in prostatic cancer diagnosis, which relates to the field of extracorporeal immune detection, and provides an ELISA detection method of an AMACR auto-antibody, that comprises the following steps: a specific antigen AMACR is used for enveloping a millipore reaction plate, blood serum to be measured is added into the millipore reaction plate, and simultaneously blank contrast, negative contrast and positive contrast are set; enzyme tag secondary antibody is added, and color reaction, termination reaction and reading are implemented; an optical absorbance is read at the 450nm position of an ELISA reader, normal blood serum is used as feminine to contrast optical absorbance Bo with the optical absorbance B of a sample to be measured, and if B is more than or equal to 2.1 times of the Bo value, the prostatic cancer diagnosis result is determined to be positive. The AMACR auto-antibody test method is applied to the prostatic cancer diagnosis and used for detecting the AMACR auto-antibody in human blood that is used as a marker in the prostatic cancer diagnosis. Compared with existing prostatic cancer diagnosis methods in the current domestic market that detect PSA concentration in human blood, the AMACR auto-antibody test method has the advantages of greatly-raised sensitivity and specificity.

Description

technical field [0001] A method for detecting AMACR autoantibodies and its application in the diagnosis of prostate cancer, the invention relates to the field of in vitro immune detection. Background technique [0002] Statistics show that 179,000 American men are diagnosed with prostate cancer each year, and the mortality rate is about 25%. There are 70,000-80,000 new cases of prostate cancer in my country every year. Advanced prostate cancer has a high mortality rate, but if detected early, prostate cancer patients can achieve a high five-year survival rate. Therefore, early diagnosis of prostate cancer is very important. The traditional diagnosis of prostate cancer relies on pathological examination, which requires taking living tissue, which brings pain and inconvenience to patients. PSA (Prostate Specific Antigen) is a protease found only in the male prostate. Currently, the PSA concentration in the blood is used as an important indicator of prostate cancer. Howeve...

Claims

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Application Information

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IPC IPC(8): G01N33/573G01N33/574
Inventor 杨静华吴斌
Owner 无锡纳生生物科技有限公司
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