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Application of anti-single-stranded DNA antibody as congenital megacolon diagnostic marker

A Hirschsprung and antibody technology, applied in the field of biomedicine, can solve the problems of impacted results, lack of abnormal intestinal innervation, and high price

Active Publication Date: 2021-07-06
GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Rectal biopsy is to directly take rectal tissue to detect whether there is a loss of ganglion cells. The accuracy rate is high, but the sampling site will affect the result
This method is interventional and invasive, and the price is very high. Generally, this method is applied to children whose barium enema is not obvious or not applicable, such as when the child has the possibility of necrotizing enterocolitis (NEC) , barium enema can lead to intestinal perforation, at this time it is not suitable to use barium enema for diagnosis, only consider rectal biopsy
Rectal manometry is to judge the abnormal innervation of intestinal nerve by detecting the lack of relaxation of the internal anal sphincter. It is only an auxiliary diagnostic method, and there are many false positives and false negatives. It cannot be used for independent detection

Method used

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  • Application of anti-single-stranded DNA antibody as congenital megacolon diagnostic marker
  • Application of anti-single-stranded DNA antibody as congenital megacolon diagnostic marker
  • Application of anti-single-stranded DNA antibody as congenital megacolon diagnostic marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Example 1. Collection and Grouping of Plasma and Tissue Samples

[0091] Plasma samples were divided into Hirschsprung children group (37 cases), other enteropathy control group (18 cases), healthy children group (30 cases), aged from 3 months to 3 years old, male sex 3 / 4 Male, disease and control groups were age and sex matched. All samples came from Guangzhou Women and Children's Medical Center, and the healthy children group was the remaining blood samples after physical examination of healthy children. The blood collection method is anticoagulant blood collection, centrifugation to separate plasma, and cryopreservation of samples. The colon tissue samples belonged to the Hirschsprung children group (36 cases), which were surgically resected diseased tissues. Other enteropathy control group (including 11 cases of colon tissue with anal stenosis and intestinal stricture stoma).

Embodiment 2

[0092] Example 2. Human autoimmune antigen chip screening and differentially expressed autoantibody analysis

[0093] Plasma from 5 cases of children with Hirschsprung’s disease (HSCR), 5 cases of plasma from children in the healthy group (HC) and 5 cases in other enteropathy (DC) groups, were used for the screening of human autoimmune antigen chips, which were provided by Guangzhou Yijin Provided by Biotechnology Co., Ltd., it includes IgG detection of more than 100 kinds of autoimmune antibodies. The original data, after subtracting the negative control, is normalized by the RLM method, and the results are analyzed by M statistics. figure 1 The cluster analysis diagram of the differential autoantibodies is shown, from which it can be seen that the anti-single-stranded DNA (ssDNA) in the plasma of children with Hirschsprung's disease is significantly higher than that in the control group (p=0.019).

Embodiment 3

[0094] Example 3. Enzyme-linked immunosorbent (ELISA) detection of anti-single-stranded DNA (ssDNA) antibodies

[0095] In order to verify the diagnostic effect of anti-single-stranded DNA (ssDNA) antibody on Hirschsprung, we collected 18 plasma samples from Hirschsprung patients (37 cases) and controls with other intestinal diseases (including children with anal stenosis and intestinal stricture fistula). example), healthy children's control (30 cases) plasma, the level of anti-single-stranded DNA (ssDNA) antibody in plasma was detected by enzyme-linked immunosorbent immunoassay (ELISA), and the detection kit was human anti-single-stranded DNA (ssDNA) antibody (ssDNA-Ab ) Enzyme-linked immunosorbent (ELISA) kit (Shanghai Zhenke Biotechnology Co., Ltd.). The result is as figure 2 It showed that the anti-single-stranded DNA (ssDNA) antibody in the plasma of Hirschsprung patients was significantly higher than that of the intestinal disease control group (p<0.01) and the contro...

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Abstract

The invention relates to the technical field of biological medicine, in particular to application of an anti-single-stranded DNA antibody as a congenital megacolon diagnostic marker. The application comprises the following steps: screening plasma of a child patient suffering from the congenital megacolon through a human autoimmune antigen chip to obtain an anti-single-stranded DNA (ssDNA) antibody highly expressed by the plasma of the child patient suffering from the congenital megacolon. The effectiveness of the anti-single-stranded DNA (ssDNA) antibody is verified in an independent sample by using an enzyme-linked immunosorbent assay (ELISA) method. The anti-single-stranded DNA (ssDNA) antibody can be used for effectively diagnosing children with congenital megacolon, and the AUC is 0.9167; the sensitivity corresponding to the optimal limit is 74.63%, and the specificity is 96.88%. The anti-single-stranded DNA (ssDNA) antibody can be used as a congenital megacolon plasma diagnosis marker for screening and diagnosing diseases, and fills the blank of congenital megacolon plasma diagnosis.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of an anti-single-chain DNA antibody as a diagnostic marker for Hirschsprung's disease. Background technique [0002] Hirschsprung disease (HSCR) is a birth defect disease with abnormal development of enteric nerves in children. The pathological mechanism is that the migration and differentiation of enteric neural crest cells into enteric neurons are impaired, resulting in the absence of enteric nerves and persistent spasm. It is one of the common congenital intestinal diseases in children. The early manifestations of Hirschsprung's disease are vomiting, abdominal distension, diarrhea, etc. Clinically, it will cause neonatal death or complications such as recurrent enteritis after surgery, refractory constipation, etc., seriously affecting the growth and quality of life of the children. [0003] Timely diagnosis and treatment of Hirschsprung can reduce the ri...

Claims

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Application Information

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IPC IPC(8): G01N33/564
CPCG01N33/564G01N2333/70571G01N2800/065
Inventor 朱云张彦左晓宇夏慧敏
Owner GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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