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Freeze-dried hemagglutination inhibition test antigen for detecting mycoplasma synoviae, antigen combination and preparation method

A technique for mycoplasma synovial bursae and hemagglutination inhibition test, applied in the field of freeze-dried hemagglutination inhibition test antigen and antigen combination for detection of mycoplasma synovial bursa Less difficult to operate, stable antigen properties, and high sensitivity

Pending Publication Date: 2020-08-14
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a MS and MG hemagglutination inhibition test antigen and its preparation method, to solve the current problems that these two antigens have poor stability and cannot be preserved for a long time

Method used

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  • Freeze-dried hemagglutination inhibition test antigen for detecting mycoplasma synoviae, antigen combination and preparation method
  • Freeze-dried hemagglutination inhibition test antigen for detecting mycoplasma synoviae, antigen combination and preparation method
  • Freeze-dried hemagglutination inhibition test antigen for detecting mycoplasma synoviae, antigen combination and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Isolation and screening of MS JS2018-4 strain

[0056] 1. Experimental materials

[0057] Mycoplasma broth basal medium (product number: CM0403B) and mycoplasma agar basal medium (product number: CM0401B) were purchased from OXOID; arginine, cysteine ​​and coenzyme I were purchased from Sangon Bioengineering (Shanghai) Co., Ltd.; pig serum was purchased from Jinyuankang Bioengineering Co., Ltd.

[0058] Preparation method of improved Frey's liquid medium: mycoplasma broth basal medium (OXOID, product number: CM0403B) 25.5g / L, glucose 3.3g / L, coenzyme Ⅰ 100mg / L, arginine 400mg / L, cysteine ​​100mg / L, 0.02% phenol red indicator, each component was added into ultrapure water, and the pH was adjusted to 7.8. Use a 0.22 μm bacterial filter to filter and sterilize, and add 15% to 20% pig serum.

[0059] The preparation method of improved Frey's solid medium: mycoplasma broth basal medium (OXOID, product number: CM0403B) 35.5g / L, autoclave at 121°C for 15min, coo...

Embodiment 2

[0067] Example 2: Preparation and application of freeze-dried MS hemagglutination inhibition test antigen

[0068] 1 Experimental materials

[0069] 1.1 Strains

[0070] The MS JS2018-4 strain was isolated, identified and preserved by the Key Open Laboratory of Livestock and Poultry Infectious Diseases of the Ministry of Agriculture of Yangzhou University, and was preserved in the China Center for Type Culture Collection (China, Wuhan, Wuhan University) with the preservation number: CCTCC M20191100.

[0071] 2 Experimental methods

[0072] 2.1 Strain cultivation

[0073] 2.1.1 Obtaining of seed bacterial liquid

[0074] Take the MS JS2018-4 isolate strain, inoculate it into fresh modified Frey's liquid medium, place it in an incubator at 37°C, and cultivate it at 220r / min for 24h, until the color of the medium changes from purple red to orange yellow. Then absorb the bacterial liquid to inoculate the freshly improved Frey's liquid medium at a ratio of 1:10, culture for 24 ...

Embodiment 3

[0121] Example 3: Preparation and application of freeze-dried MG hemagglutination inhibition test antigen

[0122] 3.1 Mass preparation of MG hemagglutination inhibition test antigen

[0123] A batch of MG hemagglutination inhibition test antigens were prepared in large quantities according to the freeze-dried MS hemagglutination inhibition test antigen preparation conditions determined in 2.1-2.4 of Example 2.

[0124] 3.2 Detection of MG hemagglutination inhibition test antigen

[0125] 3.2.1 HA titer determination

[0126] Use 2ml of PBS solution to dissolve the hemagglutination inhibition test antigen, and measure the HA titer. The results showed that the HA titers of 10 randomly selected bottles of freeze-dried antigens were all 2 8 .

[0127] 3.2.2 Specificity test

[0128] Using the method of HI test, the MG, MS, NDV, AIV, IBV positive serum and SPF chicken serum were tested for HI by using the prepared MG hemagglutination inhibition test antigen. The results are ...

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Abstract

The invention belongs to the technical field of veterinary diagnosis, and particularly relates to a freeze-dried hemagglutination inhibition test antigen for detecting mycoplasma synoviae (MS), an antigen combination and a preparation method. The freeze-dried hemagglutination inhibition test antigen for MS is prepared by the steps of selecting a mycoplasma synoviae CCTCC No.M 20191100 strain, andcarrying out seed liquid culture, enlarged culture, inactivation, concentration and freeze drying. The invention further discloses the combination of the freeze-dried hemagglutination inhibition testantigen with a Mycoplasma gallisepticum (MG) hemagglutination inhibition test antigen. The hemagglutination inhibition test antigen and the combination thereof provided by the invention are stable inproperty and easy to store. The HI detection method established by utilizing the antigen is simple and rapid to operate and low in cost, and has good sensitivity and specificity.

Description

technical field [0001] The invention belongs to the technical field of veterinary diagnosis, and in particular relates to a freeze-dried hemagglutination inhibition test antigen for detecting mycoplasma synovial bursae, antigen combination and preparation method. Background technique [0002] Mycoplasma Synoviae (MS) and Mycoplasma gallisepticum (MG) are the two most common pathogenic mycoplasmas that seriously endanger the poultry industry. MS infection can cause acute or chronic infectious diseases characterized by joint exudative bursitis and tenosynovitis, which seriously affect the production performance of laying hens and broilers, and bring serious losses to the breeding industry. The disease caused by MG-infected chickens is usually called chronic respiratory disease (Chronic respiratory disease, CRD). Difficulty, coughing and sneezing. MG-infected chickens can lead to low feed conversion rate and decreased weight gain. Infected broiler chickens will lead to increas...

Claims

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Application Information

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IPC IPC(8): G01N33/569C12N1/20C12N1/04C12R1/35
CPCG01N33/56933C12N1/20C12N1/04G01N2333/30G01N2469/20Y02A50/30
Inventor 张小荣何吉鑫吴艳涛孟林春曹永忠郭梦娇陈梦瑶谢迪张成成
Owner YANGZHOU UNIV
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