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Inert carrier Escherichia coli and its potential application

A technology of Escherichia coli and an inert carrier, applied to the inert carrier Escherichia coli and its potential application fields, can solve the problems of affecting the purification effect of epidemic diseases, and interfere with the results of detection and diagnosis, and achieve great application value and market prospects. The effect of specific bottlenecks

Active Publication Date: 2019-09-20
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The bottleneck technology for the accuracy of agglutination antigen detection needs to be improved and perfected. The root cause is that the agglutination antigens currently used in agglutination tests are all bacterial antigens, and their components are not single antigens, not single-factor components, but multiple complexes. Theoretically speaking, there are non-specific cross-reactions with bacteria of the same genus, other species, and other components of the bacterial particle antigen of the same component. In view of the working concentration of the agglutinated antigen and the number of bacteria, this drawback will inevitably affect or even significantly interfere with the detection. and diagnostic results, including animal clinical use of the agglutination test as a disease purification technology to screen pullorum-positive animals and eliminate and kill breeding birds, thereby affecting the effect of disease purification and the advancement of disease purification work

Method used

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  • Inert carrier Escherichia coli and its potential application
  • Inert carrier Escherichia coli and its potential application
  • Inert carrier Escherichia coli and its potential application

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Example 1: Isolation and identification of Escherichia coli SE1

[0023] Since October 18, 2016, 360-day-old healthy laying hens were collected from the 2nd and 4th healthy chicken flocks of the first poultry farm of Mashan Poultry Farm in Wuxi, Jiangsu, and put them in an ultra-clean table, and cleaned them with alcohol. Disinfect the surface of the chicken body, aseptically remove the liver, spleen, intestinal tract and other organ tissues of the chicken, place them in a sterile petri dish, cut up the tissues and grind them at the same time, weigh them and suck the homogenate into a sterile test tube for later use. Buffered peptone water (BPW) was weighed and autoclaved for aliquots, and the collected tissue samples were added to BPW for overnight incubation at 37°C. Take 1 mL of Escherichia coli test broth (EC broth) for enrichment culture at 37°C, and after overnight culture, draw 20 μL of enrichment solution to streak culture on MacConkey agar medium at 37°C, pick ...

Embodiment 2

[0030] Example 2: Verification that inert Escherichia coli SE1 and chicken-derived serum do not produce non-specific agglutination

[0031] The inert carrier bacteria SE1 cultivated overnight were centrifuged at 4000 rpm for 10 min at 4°C, discarded the supernatant, resuspended with sterile PBS, centrifuged and washed three times, and then resuspended to different working concentration gradients of bacteria. Before the test, the bacterial solution was mixed with a vortex instrument, and the agglutination test was performed with sterile PBS and SPF chicken serum to ensure that the bacterial solution had no self-agglutination and no non-specific agglutination. In the ultra-clean bench (20-25°C), take several pieces of ordinary glass plates with a clean surface, resuspend and wash the carrier bacteria by centrifugation and washing for 3 times with PBS pre-cooled to 4°C, and then resuspend and dilute to the specified bacterial concentration. Use a micropipette to draw a drop of ab...

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Abstract

The present invention relates to an inert carrier Escherichia coli and its potential application, the Escherichia coli isolated strain has the inert carrier property, and is expected to be developed into a carrier strain for an indirect agglutination test. The inert carrier Escherichia coli is preserved in the China General Microorganisms Collection and Management Center (CGMCC), and a preservation address is Beijing, China, a preservation number is CGMCC No. 17339, a preservation date is March 18, 2019, a classification name is Escherichia coli, and a strain code is SE1. The Escherichia coli isolated strain is from a healthy flock, the bacteria number at a higher working concentration does not cause any macroscopic agglutination reaction with various chicken serums of different genetic backgrounds, and does not generate non-specific agglutination reaction with chicken source serum, so that the Escherichia coli is called as an inert carrier Escherichia coli. The application provides an inert carrier strain and its potential application for developing a simple and rapid indirect agglutination assay.

Description

[0001] field of invention [0002] The invention belongs to the field of biotechnology detection, and relates to an inert carrier Escherichia coli, its isolation and identification and its application. In view of the fact that the number of bacteria in the working concentration of this carrier Escherichia coli does not occur in non-specific agglutination reaction of chicken source serum, and its It provides an inert carrier and has potential application prospects in the development of an indirect agglutination test method for the simple and rapid detection of antigens or infection antibodies. Background technique [0003] The development of immunology provides many technical methods for the simple and rapid detection of pathogenic bacteria in clinical practice. Among them, agglutination test is a traditional and classic rapid diagnosis method for epidemic diseases widely used in medical and veterinary clinics. After the antigen is combined with the corresponding antibody, when...

Claims

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Application Information

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IPC IPC(8): C12N1/20G01N33/569C12R1/19
CPCC12N1/205C12R2001/19G01N33/56916G01N2333/245
Inventor 朱国强杨斌羊扬夏芃芃孟霞段强德朱晓芳
Owner YANGZHOU UNIV
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