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

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...

AI Technical Summary

Problems solved by technology

Although the application of biochip technology in the field of prenatal screening has not been reported at home and abroad, there are many successful experiences in the application of diagnostic biochips in the medical field for reference.

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