30 results about "Bovine coronavirus" patented technology

The invention discloses a recombinant multi-epitope antigen of bovine coronavirus, also discloses application of the recombinant multi-epitope antigen of the bovine coronavirus in bovine coronavirus prevention ad diagnosis and belongs to the field of molecular immunology. An amino acid sequence of the recombinant multi-epitope antigen of the bovine coronavirus is a concatemer of a bovine coronavirus BCoV S protein epitope. The recombinant multi-epitope antigen, capable of causing a neutralizing antibody, of the bovine coronavirus serves as an immunogen or a vaccine, the neutralizing antibody oriented to the bovine coronavirus BCoV can be produced after the animal body is immunized, and the BCoV can be neutralized in vitro to prevent the animal body from virus infection. In addition, the recombinant multi-epitope antigen of the bovine coronavirus can also serve as a reagent for detecting a bovine coronavirus BCoV antibody or a polypeptide antibody.

The invention discloses a bovine coronavirus VPN (virtal private network) gene, encoded recombinant protein and an application thereof. A BCoV (bovine coronavirus) indirect ELISA (enzyme linked immunosorbent assay) method is established by using the recombinant protein. The bovine coronavirus VPN has the advantages of having a good antigenic epitope and high expression in prokaryotic cells, constructing a prokaryotic expression vector, successfully establishing the BCoV indirect ELISA diagnostic method by coating the antigen with the expressed recombinant protein, determining the best conditions for the method to provide a quick and easy serological differential diagnosis method for immunological detection of bovine antibodies and epidemiological investigation, and solving current drawbacks of a diagnosis method of yak coronavirus diarrhea, which is time-consuming, laborious and cannot be diagnosed quickly, and expensive expenses of biological reagents and instruments.

The invention discloses a yak coronavirus detection kit based on a constant temperature isolated fluorescent PCR platform and an application thereof; the yak coronavirus detection kit consists of a reaction buffer liquid, a fluorescent quantitative PCR reaction liquid freeze-dried tube, and positive control and negative control freeze-dried tubes. The fluorescent quantitative PCR reaction liquid freeze-dried tube is prepared by mixed freeze-drying of a Taq enzyme, a reverse transcriptase, a primer for detection, a probe and dNTPs. The kit has the advantages of short detection time (the reaction time is only 42 minutes), high specificity, high sensitivity and easy storage (storage at the temperature of 4 DEG C), is used with cooperation of a constant temperature isolated PCR instrument, issuitable for spot rapid detection of yak coronavirus, and can be widely popularized at a basic level.

The invention provides a triple fluorescent quantitative PCR kit for simultaneously detecting a bovine rotavirus, a bovine coronavirus and a bovine viral diarrhea virus and an application, and belongs to the technical field of virus detection. The triple fluorescent quantitative PCR kit comprises three groups of specific primers and three corresponding probes aiming at the bovine rotavirus, the bovine coronavirus and the bovine viral diarrhea virus. The primer and the probe for detecting the three viruses have the advantages of strong specificity and high reaction efficiency. The triple real-time fluorescent quantitative PCR kit has the characteristics of high amplification efficiency, strong specificity, stable detection effects and good repeatability, and is suitable for dairy cow diarrhea pathogen detection, regular monitoring, dairy cow diarrhea epidemiological investigation and the like. The kit disclosed by the invention can be used for simultaneously detecting three dairy cow diarrhea pathogens in a real-time fluorescent quantitative PCR reaction system, so that the detection time is saved, the detection cost is reduced, a plurality of samples can be simultaneously detected, and the detection efficiency is high.

The invention provides a bovine rotavirus fusion protein and a calf diarrhea polyvalent vaccine, and relates to the technical field of molecular biology. The bovine rotavirus fusion protein contains aVP6 fragment, and the VP6 fragment contains an amino acid sequence as shown in SEQ ID NO.4, wherein at least one loop region in the following (a)-(c) is replaced with an epitope derived from bovine coronavirus and / or an epitope derived from escherichia coli: (a) 168th-177th amino acid residues with an amino acid sequence as shown in SEQ ID NO.1; (b) 194th-205th amino acid residues with an amino acid sequence as shown in SEQ ID NO.2; and (c) 296th-316th amino acid residues with an amino acid sequence as shown in SEQ ID NO. 3. The bovine rotavirus fusion protein contains a plurality of epitopes, and a host can generate a plurality of antibodies after the host is immunized.

The present invention provides a multiplex RT-PCR / PCR method, which enables in a single assay the simultaneous detection of any combination of bovine rotavirus, bovine coronavirus, Cryptosporidium parvum, and optionally, Escherichia coli strains producing K99 pili or heat-stable enterotoxin STa.

A canine respiratory coronavirus (CRCV) that is present in the respiratory tract of dogs with canine infectious respiratory disease and which has a low level of homology to the enteric canine coronavirus, but which has a high level of homology to all bovine coronavirus strains (e.g., Quebec and LY138) and human coronavirus strain OC43.

The invention discloses a triple real-time fluorescent quantitative PCR (Polymerase Chain Reaction) primer and probe combination for simultaneously detecting BVDV (Bovine Viral Diarrhea Virus), BRV (Bovine Rotavirus) and BCV (Bovine Coronaviruses), the specificity of the primer and the specificity of the probe aim at the 5 'UTR gene segment of the BVDV, the VP6 gene segment of the BRV and the N gene segment of the BCV, and the nucleotide sequences of the primer and the probe are as shown in SEQ ID No.1-9. The invention further discloses a kit for detecting the BVDV, the BRV and the BCV. According to the invention, qualitative or quantitative detection of three viruses can be realized only by performing PCR amplification on a sample once, and the method has the advantages of simplicity and convenience in operation, strong specificity, high sensitivity, good repeatability and the like.

The invention discloses a primer set for detecting bovine norovirus and bovine coronavirus. The primer set is composed of primers for detecting bovine norovirus and primers for detecting bovine coronavirus. The core of the primer set is The nucleotide sequence is shown in SEQ ID NO.1-SEQ ID NO.4. The invention also discloses a dual RT-PCR method for detecting bovine norovirus and bovine coronavirus, the steps are as follows: (1) synthesizing the primer set for detecting bovine norovirus and bovine coronavirus; (2) extracting the primers to be tested The RNA in the sample is reverse-transcribed using the extracted RNA as a template to obtain its cDNA; (3) using the cDNA prepared in step (2) as a template, the primer set synthesized in step (1) is used for PCR amplification reaction; (4) ) to analyze PCR amplification products. The double RT-PCR detection method is simple, fast, economical and efficient, can greatly save detection time and improve detection efficiency.

The invention relates to the technical field of immune detection, and discloses a bovine coronavirus chimeric antigen and a colloidal gold immunochromatographic test paper card for detecting bovine coronavirus antibodies. The chimeric antigen sequence of the present invention is shown in SEQ ID NO:1. The present invention provides a novel bovine coronavirus chimeric antigen, which has high specificity, repeatability and accuracy when used as a bovine coronavirus antibody detection antigen, and can be applied in related immunochromatographic detection products. The results are intuitive and can be judged by the naked eye. It can quickly and specifically detect bovine coronavirus antibodies in serum, and solve the problem of rapid diagnosis of calf diarrhea in large-scale dairy farms. It has high practical value and is easy to promote and use at the grassroots level.

The invention relates to a double inactivated vaccine for preventing bovine rotavirus and bovine coronavirus and a preparation method thereof. The active components of the vaccine of the present invention include inactivated antigens of bovine rotavirus and bovine coronavirus, and the antigen components of the double inactivated vaccine can be suspended by passaged cell lines (for bovine rotavirus and bovine coronavirus) or microcarriers Cultivated (for bovine rotavirus) for virus propagation, and optimized the seedling production process. The results of vaccine safety, efficacy test and antibody tracking show that there is no adverse reaction after immunizing cattle with the dual inactivated vaccine of the present invention, and no abortion is caused after immunizing pregnant cattle. All the immunized cows produced immune protection, and the newborn calves could acquire high levels of maternal antibodies through breast milk. The results show that the vaccine of the present invention is safe and reliable, and can be used to prevent the occurrence of diarrhea in calves.

The invention discloses an ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit for bovine coronavirus. The ELISA detection kit is characterized in that purified BCoV-CD isolate recombinant pET-32a-N protein is used as coating antigen, reaction conditions are optimized, and an indirect ELISA diagnosis method for detecting bovine-serum characteristic N protein antibodies. The ELISA detection kit disclosed by the invention has the beneficial effects that escherichia coli is adopted for prokaryotic expression, the raw-material sources are wide, the preparation is easy, the standardization iseasy, and the production and detection processes are safer and more reliable, so that the ELISA detection kit is suitable for being promoted and used in basic units.

The invention discloses a triple RPA detection kit for bovine viral diarrhea virus, bovine coronavirus and bovine rotavirus, comprising SEQ.ID.NO.1 and SEQ.ID.NO.5, SEQ.ID.NO. RPA of bovine viral diarrhea virus, bovine coronavirus and bovine rotavirus composed of nucleotide sequences shown in .8 and SEQ.ID.NO.13 and SEQ.ID.NO.15 and SEQ.ID.NO.18 Detection primer set. The kit is based on the recombinase polymerase amplification method, uses cDNA formed by reverse transcription of RNA as a template to carry out amplification reaction, and uses nucleic acid gel electrophoresis to obtain detection results. The invention can be used to detect bovine viral diarrhea virus, bovine coronavirus and bovine rotavirus simultaneously, has simple operation, high sensitivity and specificity, and provides a rapid detection reagent for on-site screening of bovine diarrhea virus.

Inactivated scours vaccines for immunization and protection of bovine animals from disease caused by infection with bovine rotavirus and bovine coronavirus, which comprise and effective amount of at least one inactivated viral strain are described. Polyvalent inactivated vaccines further comprising an effective amount of an antigenic component which is protective against one or more additional pathogenic organisms or viruses are also disclosed. Said vaccines are prepared from one or more strains of rota- and coronavirus, C. perfringens Type C bacteria and E. coli bacteria, and combinations thereof. Preferably, a polyvalent inactivated vaccine is provided for parenteral administration. Passive immunity is achieved in neonatal calves via immunization of pregnant cows prior to birth.

The invention discloses a recombinant multi-epitope antigen of bovine coronavirus, also discloses application of the recombinant multi-epitope antigen of the bovine coronavirus in bovine coronavirus prevention ad diagnosis and belongs to the field of molecular immunology. An amino acid sequence of the recombinant multi-epitope antigen of the bovine coronavirus is a concatemer of a bovine coronavirus BCoV S protein epitope. The recombinant multi-epitope antigen, capable of causing a neutralizing antibody, of the bovine coronavirus serves as an immunogen or a vaccine, the neutralizing antibody oriented to the bovine coronavirus BCoV can be produced after the animal body is immunized, and the BCoV can be neutralized in vitro to prevent the animal body from virus infection. In addition, the recombinant multi-epitope antigen of the bovine coronavirus can also serve as a reagent for detecting a bovine coronavirus BCoV antibody or a polypeptide antibody.

The invention discloses a detection kit for detecting BCV and a method for applying the same to non-disease detection. The detection kit for detecting the BCV comprises an amplification primer as shown in SEQ ID NO. 3 and an amplification primer as shown in SEQ ID NO. 4. The detection kit for detecting the BCV and the application method thereof can rapidly and specifically detect the BCV.