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Rapid differential diagnosis method of riemerella anatipestifer disease and colibacillosis

A technology for Riemerella and colibacillosis, applied in the field of molecular biology, can solve the problems of time-consuming and low sensitivity, and achieve the effects of low cost, high efficiency and strong specificity

Inactive Publication Date: 2010-06-16
GUANGXI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods of bacterial isolation and identification are used to detect the mixed infection of RA and E.coli, but it is time-consuming and the sensitivity is not strong, while the single PCR detection for RA cannot be completed simultaneously for the mixed infection of RA and E.coli detection

Method used

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  • Rapid differential diagnosis method of riemerella anatipestifer disease and colibacillosis
  • Rapid differential diagnosis method of riemerella anatipestifer disease and colibacillosis
  • Rapid differential diagnosis method of riemerella anatipestifer disease and colibacillosis

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Experimental program
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Effect test

Embodiment Construction

[0058] 1. Preparation of Enrichment Medium

[0059] The enrichment medium was prepared according to the following method:

[0060] - Nacl: 0.5g;

[0061] - Peptone: 1 g;

[0062] ——Beef broth: 90mL (concentration: 0.5g / ml);

[0063] - Calf serum: 10mL (5% concentration)

[0064] Mix Nacl, tryptone, and beef broth evenly, sterilize for 15 minutes at a standard pH value of 7.2 to 7.4, at a temperature of 121°C or at a high pressure of 107.9kPa, cool the medium to room temperature, add calf serum aseptically, and mix well It can be used later or sealed and stored at room temperature for later use.

[0065] Note: The newly prepared medium should be tested for sterility and tested with known positive and negative bacteria to ensure the quality of the medium.

[0066] 2. Preparation of samples to be tested

[0067] Bacterial plate isolation: use chocolate plates to isolate bacteria from suspected sick ducks, culture them in candle jars for 24 to 48 hours, and then take the col...

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Abstract

The invention discloses a rapid differential diagnosis method of riemerella anatipestifer disease and colibacillosis. The method includes that riemerella anatipestifer 42-KD outer membrane protein gene and Escherichia coli phoA gene are selected to synthesize two pair of specificity primers used for gene amplification of the two germs, the amplified segments are respectively 809bp and 622bp, difference of the two is 187bp, and the established RA and E.coli double PCR method can rapidly, specifically and sensitively can detect and differentiate the two bacteria. The invention has the advantage that the method of the invention has lower required expense, higher efficiency and stronger specificity compared with the traditional differential diagnosis method.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a rapid differential diagnosis method for ducks infected with Riemerella and colibacillosis. Background technique [0002] Riemerella anatipestifer infection (RAI) is a contagious disease caused by Riemerella anatipestifer (RA) that mainly harms ducklings aged 2-3 weeks. The form of chronic sepsis, its pathological features are fibrinous pericarditis, perihepatitis, air sacculitis, caseous salpingitis and meningitis, often mixed with Escherichia coli infection, which brings certain difficulties to the diagnosis and treatment of the disease; the onset of the disease High rate, high mortality rate, high elimination rate, slow growth of resistant ducks, etc. cause huge economic losses to the duck industry. RA is a Gram-negative bacilli. [0003] Colibacillosis is a disease caused by certain pathogenic serotype strains of Escherichia coli (E. coli), which is a Gram-negative bacillu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12R1/19C12R1/01
Inventor 韦平尤岩岩韦天超杨宗维
Owner GUANGXI UNIV
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