Soybean peroxidase immune biochip and application of thereof to detection of serum marks during down syndrome prenatal screening

A peroxidase, biochip technology, applied in the direction of analysis, biological testing, chemiluminescence/bioluminescence, etc. by chemical reaction of materials, to achieve the effect of wide range of action, good acid-base stability, and broad application prospects

Inactive Publication Date: 2014-07-02
靖江市人民医院 +1
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  • Application Information

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Problems solved by technology

Although the application of biochip technology in the field of prenatal screening has not been reported at home and abr

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  • Soybean peroxidase immune biochip and application of thereof to detection of serum marks during down syndrome prenatal screening

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Embodiment Construction

[0030] The soybean protease used in the present invention is purchased from American Bio-Research Products Company, and the MORPH and SPTZ used in the present invention are purchased from Sigma Aldrich.

[0031] Apply the detection method of described soybean peroxidase immune biochip, the steps are:

[0032] The first step, the preparation of soybean protein enzyme-labeled antibody: prepared by the improved sodium periodate method;

[0033] The second step is the modification of the glass slide: hydroxylation, aminosilanization, and aldehydes on the surface of the glass slide;

[0034] The third step is the construction of the sandwich immune model on the surface of the biochip: coat the antibody on the surface of the glass slide, and the antibody is consistent with the enzyme-labeled antibody; block the non-specific active site; add the serum to be tested, incubate and wash; add Enzyme-labeled antibody, incubation, washing; adding luminescent substrate, CCD imaging to colle...

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Abstract

The invention discloses a soybean peroxidase immune biochip and application thereof to detection of serum marks during down syndrome prenatal screening. The application comprises the following steps: 1,preparing a soybean protein enzyme labelled antibody; 2, modifying a slide, namely performing hydroxylation, amino silanization and formylation on the surface of the slide; and 3, constructing a sandwich immune model on the surface of the biochip, namely covering the surface of the slide with the antibody, closing a non-specific active site, adding serum to be detected, performing incubation and washing, adding the enzyme labelled antibody, then performing incubation and washing again, adding a luminous substrate, and acquiring a luminous signal through CCD (charge coupled device) imaging. Four indexes, namely AFP (alpha fetoprotein), HCG (human chorionic gonadotropin), uE3 and PAPP (pregnancy associated plasma protein)-A, are selected as down syndrome serum marks. The SBP (soybean peroxidase) is wide in substrate working range, high in heat resistance, high in acid-alkaline stability and wide in pH adaptation range. An enhancer is added into a Luminol-H2O system, so that a chemical luminous signal can be amplified by nearly 100 times, and the detection sensitivity is improved.

Description

technical field [0001] The invention relates to the field of biochip technology, in particular to the application of biochip technology in prenatal Down's syndrome screening and the use of a novel labeling enzyme (SBP) and an ultra-high sensitivity chemiluminescent detection method. Background technique [0002] Now medical testing technology is developing in the direction of standardization, intelligence and intensification. Biochip technology is the leader of this trend. It is a method to orderly solidify a large number of biological macromolecules such as nucleic acid fragments, polypeptide molecules, antigens, antibodies and other biological samples on glass slides and silicon chips through microprocessing technology and microelectronics technology. Sheets, glass sheets, plastic sheets, polyacrylamide gels, nylon membranes and other solid-phase media surface to form a dense two-dimensional molecular arrangement, and then hybridize with the target molecules in the labeled...

Claims

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Application Information

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IPC IPC(8): G01N33/96G01N33/535
CPCG01N33/689G01N21/76G01N33/535G01N2800/387
Inventor 薛艳春苏娟车彦军王建江毛世琴朱伟田亦平蒋镌翟丽芬陶岚刘松琴沈丽吴慧萍
Owner 靖江市人民医院
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