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Detection kit and method for pig intestinal injuries

A detection kit and a technology for intestinal damage, applied in the biological field, can solve the problems of complex process, time-consuming and laborious cost, and complicated operation of detection and analysis methods, and achieve the effect of accurate determination.

Active Publication Date: 2018-06-29
HUBEI HAOHUA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the prior art, although there are methods for evaluating the intestinal injury through intestinal anatomy, clinical symptoms, microscopic observation of mucosal morphology and structure, and molecular biological detection and analysis of genes and protein expressions, the existing detection and analysis methods operate It is cumbersome, the process is complicated, the professional level is too high, time-consuming, labor-intensive and costly, it is difficult to achieve rapid, safe, stable and reliable detection and evaluation of intestinal damage

Method used

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  • Detection kit and method for pig intestinal injuries
  • Detection kit and method for pig intestinal injuries
  • Detection kit and method for pig intestinal injuries

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 2

[0109] Example 1 Diamine oxidase detection

[0110] (1) Collection of pig blood and separation of plasma samples: blood was collected from the anterior vena cava of the piglets to be tested and placed in an anticoagulation tube. After standing for 20 minutes, centrifuged at 3500 rpm for 15 minutes and then the supernatant was taken as the sample to be tested. .

[0111] (2) Sample addition: Blank holes (no enzyme-labeled reagent and HRP-labeled antibody are not added, the sample is replaced with an equal dose of enzyme-free water), sample wells to be tested (add sample to be tested), negative control In the wells (add the pig intestinal injury negative standard plasma) and the positive control well (add the pig intestinal injury positive standard plasma), HRP-labeled diamine oxidase antibody is added to each hole. Add 40μl of sample or standard plasma to the microtiter plate coated with diamine oxidase antibody, add 40μl of enzyme-free water to the blank well, and then add horsera...

Embodiment 2

[0118] Example 2 Citrulline Detection

[0119] (1) Collection of pig blood and separation of plasma samples: blood was collected from the anterior vena cava of the piglets to be tested and placed in an anticoagulation tube. After standing for 20 minutes, centrifuged at 3500 rpm for 15 minutes and then the supernatant was taken as the sample to be tested. .

[0120] (2) Sample addition: Blank holes (no enzyme-labeled reagent and HRP-labeled antibody are not added, the sample is replaced with an equal dose of enzyme-free water), sample wells to be tested (add sample to be tested), negative control In the wells (adding the pig intestinal injury negative standard plasma) and positive control wells (adding the pig intestinal injury positive standard plasma), HRP-labeled citrulline antibody is added to each hole. Add 40μl of sample or standard plasma to the microtiter plate coated with diamine oxidase antibody, add 40μl of enzyme-free water to the blank well, and then add 10μl of horser...

example 1

[0128] 28 healthy weaned piglets were selected from the pig farm and randomly divided into four groups: blank group (no irritation, as a negative control for intestinal injury), LPS group (lipopolysaccharide LPS intraperitoneal injection), PEDV group (oral swine prevalence Diarrhea virus (PEDV), STa group (orally fed with E. coli type I heat-resistant enterotoxin STa), stimulated in the LPS group for 3 hours, and 24 hours after inoculation in the PEDV and STa groups, using anticoagulant vacuum blood collection tubes and blood collection needles Blood was collected from the anterior vena cava of the test pig, and turned upside down to mix the anticoagulant and blood evenly. After standing for 20 minutes, centrifuged at 3500r / min for 15 minutes, and then the supernatant was taken for testing.

[0129] The OD value of diamine oxidase measured according to Example 1 is shown in Table 1, and the identification result is shown in Table 2. The citrulline absorbance value measured accordi...

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Abstract

The invention discloses a detection kit and method for pig intestinal injuries. The detection kit for the pig intestinal injuries comprises a microplate coated with a pig diamine oxidase and citrulline antibody, an HRP (horseradish peroxidase) labeled diamine oxidase and citrulline antibody, a TMB substrate solution and positive standard plasma and negative standard plasma for the pig intestinal injuries. The method for detecting and evaluating the intestinal injuries is simple to operate and has low professional level requirement and low cost, and the intestinal injuries can be detected and evaluated rapidly, safely, stably and reliably.

Description

Technical field [0001] The invention relates to the field of biotechnology, in particular to a detection kit and a detection method for pig intestinal injury. Background technique [0002] Diamine oxidase is a highly active intracellular enzyme in the upper villi of the small intestinal mucosa of humans and mammals. It plays a role in the metabolism of histamine and a variety of polyamines. Its activity is closely related to the nucleic acid and protein synthesis of mucosal cells and can reflect The integrity and degree of damage to the intestinal mechanical barrier. Diamine oxidase is highly expressed in dividing cells. Necrosis of intestinal mucosal cells fall off into the intestinal lumen, which reduces the activity of diamine oxidase in the intestinal mucosa, and increases the activity of diamine oxidase in intestinal contents; diamine oxidase enters the intercellular space of the intestines, which increases the activity of diamine oxidase in plasma , The change of plasma d...

Claims

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Application Information

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IPC IPC(8): G01N21/31C12Q1/26
CPCC12Q1/26G01N21/314
Inventor 侯永清吴涛丁斌鹰易丹
Owner HUBEI HAOHUA BIOTECH
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