Specific primer and method for detecting bovine derived proteus

A Proteus mirabilis, specific technology, applied in the field of microorganisms, can solve problems such as unfavorable diagnosis of etiology, inappropriate routine detection, etc., and achieve the effects of avoiding cumbersome operations, short detection time, and high sensitivity

Pending Publication Date: 2019-12-10
SICHUAN AGRI UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] The traditional bacterial identification method is mainly based on the classification of bacterial physiological and biochemical characteristics, which requires a lot of time to interpret the results of physiological and biochemical reactions, which is not conducive to timely diagnosis of the cause, finding the pathogen, and controlling the spread of the disease
However, after the 16SrRNA universal primer is amplified, it needs to be sequenced to determine the bacterial species, which is not suitable for routine detection

Method used

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  • Specific primer and method for detecting bovine derived proteus
  • Specific primer and method for detecting bovine derived proteus
  • Specific primer and method for detecting bovine derived proteus

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Embodiment Construction

[0033] The technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0034] In this embodiment, for the virulence protein gene (rsbA gene) of Proteus mirabilis, a specific primer for detecting bovine Proteus mirabilis with strong specificity and high sensitivity is designed, including forward primer and Reverse primer, specifically:

[0035] Forward primer F: 5'-ATGCGCACACTGACCCAATTA-3';

[0036] Reverse primer R: 5'-CTGATCGCGTCCTTCAAGCC-3';

[0037] The specific primers are shown in SEQ ID NO.1-2.

[0038] Such as figure 1 Shown, a kind of method of detecting bovine source Proteus mirabilis of the present invention may further comprise the steps:

[0039] S1. Extracting the genomic DNA of the sample to be tested; including the following sub-steps:

[0040] S11. Take 100-200 μL of the base solution of the sample to be tested that has been incubated in a constant temperature incuba...

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Abstract

The invention discloses a specific primer and a method for detecting bovine derived proteus. The primer comprises a forward primer and a reverse primer. Specifically, the forward primer is F:5'-ATGCGCACACTGACCCAATTA-3', and the reverse primer is R:5'-CTGATCGCGTCCTTCAAGCC-3'. The specific primer is disclosed in SEQ ID NO.1-2. The detection method comprises the following steps of: S1: extracting thegenome DNA (deoxyribonucleic acid) of a sample to be detected; S2: using the specific primer for detecting the bovine derived proteus to carry out PCR (polymerase chain reaction) amplification on thegenome DNA of the sample to be detected; and S3: carrying out gel electrophoresis detection, carrying out photographing detection under a gel imaging system, if a DNA specific band of 306bp is in thepresence, determining that the sample contains the proteus, and otherwise, judging that the sample does not contain the proteus. The primer provided by the invention can be used for the PCR detectionof the proteus, has the characteristics of short detection time, low cost, high detection result specificity and high practicality, and a result is easy in interpretation. The detection method established by the invention utilizes a PCR technology, and therefore, the detection method has the characteristics of being convenient in operation, quick, high in sensitivity, high in specificity and thelike.

Description

technical field [0001] The invention belongs to the technical field of microbes, in particular to a specific primer and method for detecting bovine Proteus mirabilis. Background technique [0002] Proteus mirabilis (Proteus) is a member of the Enterobacteriaceae Proteus genus. It is a non-capsulated, non-spore-forming, flagellated Gram-negative bacilli. It is famous for its unique periodic clustering movement. Microscopic observation shows multiple Shape, mostly rod-shaped, less spherical. Proteus mirabilis is widely distributed in nature, mainly in sewage, soil, and feces. Cause enteritis, pneumonia, cystitis, kidney stones and other diseases, and has become the main cause of complicated urinary tract infection (CAUTI). With the continuous expansion of the scale and density of beef cattle breeding in China in recent years, factors such as the deterioration of the feeding environment and long-distance transportation often lead to the decline of the body resistance of the c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/10C12N15/11C12R1/37
CPCC12Q1/686C12Q1/689C12Q2565/125
Inventor 马志宇才冬杰左之才谢跃任志华马晓平叶刚余树民苟丽萍王娅
Owner SICHUAN AGRI UNIV
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