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Application of salmonella pullorum secreted protein SpiC

A Salmonella pullorum, secretion technology, applied in the field of Salmonella pullorum secretory protein SpiC, can solve the problems that there are no reports on the immune detection of Salmonella pullorum infection, and achieve strong specificity, good sensitivity and specificity, sensitivity and highly specific effect

Active Publication Date: 2013-07-10
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
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  • Application of salmonella pullorum secreted protein SpiC
  • Application of salmonella pullorum secreted protein SpiC
  • Application of salmonella pullorum secreted protein SpiC

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Embodiment 1

[0069] 1. Test method

[0070] 1. Construction of recombinant plasmids and prokaryotic expression

[0071] 1.1 Amplification of the target gene spiC The gene sequence of spiC was determined according to NCBI search. Use Primer5 software to design primer P1:5'-TA ATGCTGGCAGTTTTAAAAAGGCATT-3' (SEQ ID NO: 3)

[0072] P2:5'-TA TTATACCCCACCCGAATAAAGTTTATG-3' (SEQ ID NO: 4), restriction sites XhoI and BamHI. The polymerase chain reaction (PCR) is carried out with primer P1 and primer P2, and the genomic DNA of the wild strain of Salmonella pullorum is used as the DNA template. The template can be obtained by cultivating the isolated wild strain of Salmonella pullorum and extracting its genomic DNA by conventional methods. The wild strain of Salmonella pullorum can be isolated from Salmonella-infected chicken food or chicken manure according to the existing technology or directly purchased from the China Microbial Strain Resource Bank. Reaction system: 10× buffer 2.5 μL, dNTP ...

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Abstract

The invention provides application of salmonella pullorum secreted protein SpiC. The salmonella pullorum secreted protein SpiC can be taken as a biological marker for diagnosis when salmonella infects a host, and is used for preparing materials for pullorum disease immunodetection or materials for diagnosis. According to the invention, the protein is further utilized to establish materials applied to pullorum disease ELISA (enzyme-linked immuno sorbent assay) of a SpiC antibody generated in the host, and a pullorum disease ELISA method is correspondingly given. The protein-based ELISA method for detecting the SpiC antibody generated in the host is good in specificity and high in flexibility, and can be applied to the evaluation of organism immune states and the research relevant to infectious disease diagnosis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of the secretory protein SpiC of Salmonella pullorum pullorum. Background technique [0002] Pullorum pullorum is an infectious disease caused by Salmonella pullorum pullorum, which occurs all over the world and is one of the most serious diseases that endanger the chicken industry. Its excrement is an important medium of transmission, and it can also be transmitted vertically through eggs. Chickens suffering from pullorum do not eat feed, are afraid of cold, curl up their bodies, droop their wings, feel depressed or lethargic, discharge white sticky or light yellow, light green loose stools, sometimes the anus is closed by hardened feces, and have difficulty breathing. Adult chickens have no clinical symptoms, and a small number of seriously infected sick chickens are listless, with yellow-green or egg white-like loose stools. The main lesions can be seen in the liver, sple...

Claims

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Application Information

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IPC IPC(8): G01N33/68
Inventor 焦新安耿士忠潘志明刘欢孙林
Owner YANGZHOU UNIV
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