38 results about "Bovine rotavirus" patented technology

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.

The present invention relates to a bovine rotavirus recombinant protein antigen and a preparation method thereof, in particular to a bovine rotavirus recombinant VP6 protein antigen and a preparation method thereof. The bovine rotavirus recombinant VP6 protein antigen resolves the following problems: at present, the method for diagnosing bovine rotavirus diarrhea needs a great deal of time and labor and cannot rapidly diagnose bovine rotavirus diarrhea; the process is complex; and expensive molecular biology reagents and corresponding instruments need to be used. The bovine rotavirus recombinant VP6 protein antigen is expressed by colibacillus converted by recombinant vector plasmids. The preparation method includes the following steps: (1) RCR amplification of VP6 gene; (2) acquisition of recombinant plasmids; (3) acquisition of recombinant bacteria; (4) inducible expression of recombinant VP6 protein antigen; (5) purification. The present invention provides a diagnosis antigen for the serodiagnosis of bovine rotavirus diarrhea, and the diagnosis antigen has extremely strong specificity and accuracy. The diagnosis method, which uses the bovine rotavirus recombinant VP6 protein antigen and adopts fluorescent antibody test and serological tests such as enzyme linked immunosorbent assay, has the advantages of time and labor saving and rapidness and does not need expensive reagents and instruments.

The invention relates to the field of virus detection, and particularly relates to a kit, a primer and a probe for simultaneously detecting bovine rotavirus, bovine rotavirus and bovine coronavirus. The nucleotide sequences of the primer and the probe in the kit are shown as SED ID NO: 1-SEQ ID NO: 9. The kit has the technical advantages of being easy and convenient to operate, high in specificity, high in sensitivity, good in repeatability and capable of simultaneously achieving qualitative detection and accurate quantification of BVDV, BRV and BCoV.

The invention discloses a primer, a probe and a reagent kit for RPA (recombinase polymerase amplification)-side flow chromatography detection of BRV (bovine rotavirus). The reagent kit contains primerprobe liquid for detecting the BRV through RPA technology, the forward primer sequence of the primer is as shown in SEQ ID No. 1, the reverse primer sequence is as shown in SEQ ID No. 2, and the probe sequence is as shown in SEQ ID No. 3. According to primer probe groups used in the invention, the RPA has a good amplification effect and strong strip specificity, does not have cross reaction withother viruses, has strong positive reaction in a detection area, and improves the sensitivity of detection. RPA-side flow chromatography technology is firstly adopted to establish a method for quick detection of the BRV, and by evaluation of specificity and sensitivity, the method can be used for clinical on-site detection, thereby providing a sensitive and reliable method for the on-site detection of the BRV.

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 bovine rotavirus, coronavirus, and viral diarrhea virus multi-connected RT-PCR detection method, which is characterized in that according to the detection method, RNA of a to-be-detected sample is taken as an RNA template, and a specificity upstream primer and a specificity downstream primer designed according to hereditary characteristics of the bovine rotavirus, coronavirus, and viral diarrhea virus are taken as a primer to perform RT-PCR proliferation to obtain a proliferation product. Then, electrophoresis detection is performed on the proliferation product with 1.5% agarose gel, and a result is recorded; according to the results of the electrophoresis detection, whether the to-be-detected sample contains the bovine rotavirus, coronavirus, and viral diarrhea virus is determined. The bovine rotavirus, coronavirus, and viral diarrhea virus detection method achieves has the beneficial effect of accurately, rapidly and efficiently distinguishing and diagnosingthe three viral disease with RT-PCR.

The invention discloses a detection agent of bovine rotavirus and bovine coronavirus, which comprises two pairs of specific primers capable of amplifying a bovine rotavirus VP7 gene and a bovine coronavirus N gene. The invention also discloses a preparation method, which comprises the following steps of: selecting conserved sequences of the bovine rotavirus VP7 gene and the bovine coronavirus N gene; and designing and synthesizing the specific primers. The invention also discloses an application method which comprises the steps of: processing a sample; extracting RNA in an excrement sample; carrying out a jointed RT-PCR (Reverse Transcriptase Polymerase chain Reaction); and judging a result. Through the jointed RT-PCR, the detection agent can be used for rapidly and effectively detecting the BRV and the BCoV simultaneously so that the detection cost and the detection time are greatly reduced, has better specificity and sensitivity, and has the advantages of saving time and saving labor.

The present invention provides vaccine compositions for protection against human rotaviral disease designed for use in particular areas of the world. Human×bovine reassortant rotavirus comprising each of the four clinically most important VP7 serotypes of human rotavirus are combined with other VP7 serotypes typically found in the area of interest into a multivalent formulation which provides a high degree of infectivity and immunogenicity. Methods and an administration protocol for producing an immunogenic response without producing an increased risk of intussusception are also provided.

The invention discloses a multi-nano-PCR primer group for detecting bovine rotavirus (BRV), bovine parvovirus (BPV) and a bovine viral diarrhea virus (BVDV) and application of the multi-nano-PCR primer group. The primer group contains dual-priming oligonucleotide (DPO) primer pairs used for detecting the BRV, the BPV and the BVDV correspondingly. A multi-DPO-nano-PCR detection method capable of simultaneously detecting the BRV, the BPV and the BVDV is established by combining DPO primers with a nano PCR technology. Compared with a conventional PCR method, the multi-DPO-nano-PCR detection method is time-saving and labor-saving, can quickly, accurately and specifically detect pathogens, and achieves high sensitivity on the basis of ensuring specificity. By providing the multi-nano PCR primergroup, the new method is provided for diagnosis of early infection and inapparent infection of the BRV, the BPV and the BVDV, a reliable technical means is provided for detection of clinical mixed infection of the BRV, the BPV and the BVD, and technical support is provided for epidemic disease testing, screening purification and comprehensive prevention and control.

The invention discloses a triple RPA (recombinase polymerase amplification) detection kit for a bovine viral diarrhea virus, a bovine coronavirus and a bovine rotavirus. The triple RPA detection kit comprises RPA detection primer groups for the bovine viral diarrhea virus, the bovine coronavirus and the bovine rotavirus, wherein the RPA detection primer groups are respectively composed of nucleotide sequences shown as SEQ ID NO.1 and SEQ ID NO.5, SEQ ID NO.8 and SEQ ID NO.13, and SEQ ID NO.15 and SEQ ID NO.18. The kit is based on a recombinase polymerase amplification method, cDNA formed by RNA reverse transcription is used as a template for amplification reaction, and a detection result is obtained by nucleic acid gel electrophoresis. The kit can be used for simultaneously detecting the bovine viral diarrhea virus, the bovine coronavirus and the bovine rotavirus, is simple to operate, has relatively high sensitivity and specificity, and provides a rapid detection reagent for on-site screening of the bovine diarrhea virus.

The invention provides a visual multiplex fluorescent LAMP detection method which can detect bovine rotavirus (BRV) and enterotoxigenic escherichia coli (ETEC). According to the detection method, twofluorophores are introduced into an LAMP method, a result is directly judged by virtue of colors of a reaction product, for example, red is namely bovine diarrhea caused by the BRV, and green is namely bovine diarrhea caused by the ETEC. The detection method provided by the invention has the advantage that the two viruses can be identified and detected at the same time through once LAMP reaction in one reaction tube. The primer designed by the invention is high in sequence sensitivity, and each reaction can detect 100 copied mixed templates at least; and specificity is good, and a target genecan be efficiently amplified; and clinical detection effect is good. The detection method provided by the invention is convenient and rapid, does not need to use an expensive instrument, is low in cost and can realize field pathogeny detection.

The invention relates to a bovine rotavirus, a bovine coronavirus combined inactivated vaccine and a preparation method thereof. The active ingredient of the vaccine of the present invention comprisesinactivated antigen of bovine rotavirus and bovine coronavirus, and the antigenic component of the combined inactivated vaccine can be subjected to virus propagation through a continuous cell line (aiming at bovine rotavirus and bovine coronavirus) or microcarrier suspension culture (aiming at bovine rotavirus), and a seedling process and the like can be optimized. The vaccine safety, effectiveness test and antibody tracking results show that no adverse reaction is generated after immunization of cattle with the combined inactivated vaccine of the present invention, and no abortion is inducedafter immunization of pregnant cattle. Immunization of cattle produces immune protection, and newborn calves can obtain high levels of maternal antibodies through breast milk. The results show that the vaccine of the invention is safe and reliable and can be used for preventing the generation of yak diarrhea.

The invention relates to the field of virus detection, and in particular relates to a kit, primer and probe for detecting bovine rotavirus. The invention provides a kit for detecting bovine rotavirus;the kit contains a nucleic acid amplification reagent; the nucleic acid amplification reagent contains a primer and a probe for detecting bovine rotavirus; the primer comprises an upstream primer asshown in SEQ ID NO:1 and a downstream primer as shown in SEQ ID NO:2; and the probe comprises a probe as shown in SEQ ID NO:3. The kit provided by the invention has the technical advantages of high sensitivity and high specificity.

The invention provides a bovine rotavirus recombinant VP7 protein antigen and a preparation method thereof and relates to a protein antigen and the preparation method thereof, and the bovine rotavirus recombinant VP7 protein antigen can be used for solving the problems that the existing bovine rotavirus detection method cannot simultaneously meet the requirements of equipment and operation simplification, good specificity and large-scale clinical popularization. The bovine rotavirus recombinant VP7 protein antigen is coded by 644bp basic groups and has a size of 40kDa. The preparation method comprises the following steps: 1, extracting genome RNA of a bovine rotavirus BRV-DQ strain; 2, carrying out reverse transcription, PCR (Polymerase Chain Reaction) amplification and purification on the genome RNA and then connecting the genome RNA with a pMD18-T carrier so as to obtain pMD18-T-VP7 plasmids; 3, simultaneously carrying out enzyme digestion on pMD18-T-VP7 and pET30a, connecting and converting into host bacteria so as to obtain recombinant bacteria pET30a-VP7/BL21; 4, carrying out inducible expression on recombinant protein; and 5, purifying the recombinant protein so as to obtain the bovine rotavirus recombinant VP7 protein antigen. The bovine rotavirus recombinant VP7 protein antigen is applied to the field of detection of bovine rotaviruses.

The invention provides a VP7 gene-based bovine rotavirus qPCR detecting kit, comprising the components such as a specific primer of a conserved sequence of a VP7 gene, reverse transcriptase and heat-resistant DNA polymerase (Taq enzyme); the detected highest dilution degree can be up to 10<-5> to 10<-4>; the linear range of quantitative detection can be up to 10<-1> dupes/mu L; the VP7 gene based bovine rotavirus qPCR detecting kit does not have any amplification to other viral nucleic acids, bacteria and negative control groups; and the detecting kit is rapid, sensitive and specific and can be used for quantitatively detecting bovine rotavirus in various samples such as feces and blood.