93 results about "Porcine rotavirus" patented technology

The invention discloses a colloidal gold immunochromatographic test strip for detecting wild-type classical swine fever virus, which consists of water absorbent paper (1), a cellulose nitrate membrane (2), a colloidal gold pad (3), a sample pad (4) and a support (5), wherein the cellulose nitrate membrane contains a detection line which is formed by coating monoclonal antibody HQ06 of anti-classical swine fever virus E2 protein and a quality control line which is formed by coating rabbit anti-mouse IgG antibody; and the colloidal gold pad is combined with colloidal gold-labeled monoclonal antibody 6E10 of the anti-classical swine fever virus E2 protein. The test strip does not react with C-strain of classical swine fever virus, bovine viral diarrhea virus, porcine reproductive and respiratory syndrome virus, transmissible gastroenteritis virus, porcine epidemic diarrhea virus, porcine rotavirus, pseudorabies virus, porcine parvovirus and porcine circovirus type 2, and can accurately and sensitively identify the wild-type classical swine fever virus, thereby having good specificity, sensitivity and repeatability.

The invention provides a triple live vaccine for a swine transmissible gastroenteritis virus, a swine epidemic diarrhea virus and a swine rotavirus and a preparation method thereof. The content of the three viruses is not less than 107.5 TCID50 (Tissue Culture Infectious Dose 50)/mL, and the volume ratio is 1:1:1. The triple live vaccine provided by the invention solves the problem that a multiple vaccine for effectively preventing and treating such three diseases as swine transmissible gastroenteritis, swine epidemic diarrhea and the swine rotavirus is not available on the current market, and especially realizes the prevention and control on the swine rotavirus. Compared with the existing method of inoculating with three simplex vaccines to prevent such three transmissible diseases, the triple live vaccine provided by the invention is economical to use, simplifies the immunization procedure and lowers the epidemic prevention cost, thereby providing a new simple and convenient immunization way for farms in China.

The invention provides a method for preparing triple vaccine for preventing porcine transmissible gastroenteritis, porcine epidemic diarrhea and porcine rotavirus. The method comprises the following steps: inoculating a host-cell line with a 90 percent grown monostratum against a porcine transmissible gastroenteritis virus, a porcine epidemic diarrhea virus and a porcine rotavirus respectively, and adding a cell maintenance media into the host-cell lines respectively to be cultured at 37 DEG C; after cytopathic effect reaches over 75 percent, collecting viruses to be stored at 20 DEG C below zero for standby; mixing the viruses according to 10 TCID50 in 1:1:1, and simultaneously adding Freund's complete adjuvant and immunopotentiator into the mixture to inactivate the mixture by formaldehyde at 37 DEG C for 24 hours; and adding an oil adjuvant into the mixture to prepare a vaccine of water-oil-water preparation. The method can be used for preparing the triple vaccine for preventing the porcine transmissible gastroenteritis, the porcine epidemic diarrhea and the porcine rotavirus so as to solve the problem that the diseases do not have an effective medicine to treat currently.

The invention provides a detection kit containing two anti porcine epidemic diarrhea virus monoclonal antibodies PEDV-McAB1 and PEDV-McAB2, two anti porcine transmissible gastroenteritis virus monoclonal antibodies and two anti porcine rotavirus monoclonal antibodies; the kit can be applied for simultaneous detection of porcine epidemic diarrhea viruses, porcine transmissible gastroenteritis viruses and/or porcine rotaviruses with non-diagnostic purpose, moreover, the detection sensitivity of the kit for simultaneous detection of two or three kinds of viruses is higher than that of single detection of one kind of virus, and false positive results are avoided.

The invention relates to a porcine rotavirus delta VP8* subunit recombinant protein and an encoding gene of the protein. The invention further provides a recombinant protein formed after increasing tetanus toxin T cell epitope P2 into the recombinant protein, and an encoding gene. The delta VP8* protein is 64th-site to 223th-site amino acid in the VP8* and can effectively stimulate an organism to produce specific serum antibody, humoral immune response is good, the problem that the VP4 gene can not conduct prokaryotic expression due to overlarge fragment can be overcome, the protein can be expressed as a soluble protein in vitro, so that the problem that the VP8* is expressed as an inclusion body in vitro can also be overcome; the T cell epitope P2 (830th-site to 844th-site amino acid of TT) in the tetanus toxin is induced into the delta VP8* subunit recombinant protein, so that the immunity efficacy of the protein can be greatly improved, the faster and stronger neutralizing antibody titer can be induced, and high-titer rotavirus cross neutralizing antibody can also be induced.

The invention discloses a multiplex PCR primer group for detecting porcine epidemic diarrhea virus, porcine transmissible gastroenteritis virus and porcine rotavirus simultaneously, and belongs to the field of virus detection. The primer group comprises three pairs of primers, wherein the primer sequences of the first pair of primers used for detecting the porcine epidemic diarrhea virus are respectively SEQ ID NO:1 and SEQ ID NO:2, the primer sequences of the second pair of primers used for detecting the porcine transmissible gastroenteritis virus are respectively SEQ ID NO:3 and SEQ ID NO:4, and the primer sequences of the third pair of primers used for detecting the porcine rotavirus are respectively SEQ ID NO:5 and SEQ ID NO:6. Through the application of the sequences of the primers, different strains of the porcine epidemic diarrhea virus, the porcine transmissible gastroenteritis virus and the porcine rotavirus can be detected simultaneously through the multiplex PCR method, and the detection results of the porcine epidemic diarrhea virus, the porcine transmissible gastroenteritis virus and the porcine rotavirus are masculine, while the defection results of other common pig-derived viruses are feminine, in conclusion, the primer group is strong in specificity, and good in repeatability; the PCR detection is carried out after the virus cDNA is subjected to gradient dilution, which shows that the sensitivity of the primers is high.

The invention discloses a kit for detecting a porcine epidemic diarrhea virus, a porcine transmissible gastroenteritis virus and a porcine rotavirus. The gene detection kit disclosed by the invention can accurately and effectively detect the porcine epidemic diarrhea virus, the porcine transmissible gastroenteritis virus and the porcine rotavirus, and is strong in specificity, high in sensitivity, short in time consumption, fast in detection and good in application prospects.

The invention discloses a DPO (Dual Priming Oligonucleotide) primer group for porcine epidemic diarrhea virus, porcine transmissible gastroenteritis virus and porcine rotavirus detection and application of the DPO primer group. Aiming to a TGEV-N gene, a PEDV-N gene and a PRoV-VP7 gene, a pair of DPO primers respectively, and a multiplex DPO real-time RT-PCR detection method of a porcine epidemicdiarrhea virus, a porcine transmissible gastroenteritis virus and a porcine rotavirus is established. Results show that a detection limit of the method is 5.3*100 copies/microlitre, target gene fragments can be efficiently amplified within an annealing temperature range from 40 DEG C to 65 DEG C, and shown that the method is wide in annealing temperature range; and meanwhile, the DPO primers are strong in specificity, and no non-specific amplification is generated in the PCR process. The multiplex DPO real-time RT-PCR detection method provided by the invention is simple to design, strong in specificity and high in sensitivity and provides a new technological means to rapid and accurate detection of three porcine viral diarrhea infective pathogens of TGEV, PEDV and PRoV.

The invention discloses a quintuple RT-PCR detection method and kit for porcine epidemic diarrhea viruses, porcine transmissible gastroenteritis viruses, porcine rotaviruses, porcine sapoviruses and porcine kobuviruses. The kit comprises ten specificity amplification primers. In the using process of the kit, total RNA of a sample to be detected is subjected to reverse transcription to become cDNA through a 6-basic-group random primer, a detection reaction system in the kit is used for RCR amplification with the cDNA as a template, and whether the sample to be detected is infected with one kind of pathogens or is under mixed infection is determined according to different sizes of amplified PCR segments of different pathogens. On the condition of ensuring specificity and sensitivity, the kit has the advantages of being easy and quick to operate and lowering detection cost and labor intensity, and is suitable for field sample detection.

The present invention relates to a test strip which is used for testing a kind of or a plurality of kinds of disease antibodies of porcine virus diarrhea; the test strip comprises a supporting layer and a reaction reagent carrier absorbing layer which is pasted on the supporting layer; the reaction reagent carrier absorbing layer comprises a testing end fiber layer, a fiber layer of absorbing gold-labeled SPA protein or gold-labeled antigen which corresponds to the antigen to be tested, and a cellulose layer, which are arranged at the sample end in sequence, and an absorbent material layer which is positioned at the handle end; the cellulose layer contains one, two or three testing print(s) which are printed by anyone, any two or three of the purified transmissible gastroenteritis virus TGEV solution, the porcine epidemic diarrhea virus PDEV solution and the porcine rotavirus RV, and the cellulose layer also contains the contrast prints which are printed by anyone, any two or three of the anti-SPA protein IgG solution of the sheep or the rabbit or the anti-TGEV, anti-PDEV, anti-RV IgG solution of the sheep or the rabbit; and the invention provides a test strip for testing a kind of or a plurality of kinds of disease antibodies of the porcine virus diarrhea, which has the advantages of accurate and rapid detection, convenient operation and low costs.

The invention relates to a detection primer and reagent kit of four RT-PCR of porcine epidemic diarrheaviruses, transmissible gastroenteritis of swine, porcine rotaviruses and porcine deltacoronaviruses, and belongs to the technical field of molecular detection. The fourfold RT-PCR detection primer comprises detection primers PEDV F and PEDV R of porcine epidemic diarrhea viruses, detection primers TGEV F and TGEV R of the transmissible gastroenteritis of swine, detection primers PoRV F and PoRV R of the porcine rotaviruses and detection primers PDCoV-F and PDCoV-R of the porcine delta coronaviruses. The primer is high in specificity, has repeatability, is high in sensitivity and is high in clinical reliability.

The invention discloses a gene chip and kit for detecting porcine epidemic diarrhea virus, transmissible gastroenteritis virus and porcine rotavirus. The gene chip disclosed by the invention comprises a solid phase carrier and a probe fixed on the solid phase carrier, wherein the probe comprises any one or two oligonucleotide fragments shown in SEQ ID NO.1 or 2, any one or two oligonucleotide fragments shown in SEQ ID NO.3 or 4, and any one or two of oligonucleotide fragments shown in SEQ ID NO.5 or 6. The kit disclosed by the invention comprises the gene chip and reagents for amplifying genes of porcine epidemic diarrhea virus, transmissible gastroenteritis virus and porcine rotavirus. The gene chip and detection kit disclosed by the invention can accurately and effectively detect the porcine epidemic diarrhea virus, transmissible gastroenteritis virus and porcine rotavirus and are good in specificity, high in sensitivity, low in time consumption, high in detection speed and good in application prospects.

The invention discloses a breeding method for breeding pigs, and relates to the technical field of pig breeding. In the breeding pig breeding process, the breeding pigs in different phases such as piglet phase, reserve swine phase and sow phase are subjected to immunization by various vaccines in different time stages. According to the breeding method, a normative immunization program is provided, so that swine fever, foot-and-mouth diseases, pseudorabies, porcine reproductive and respiratory syndrome, parvovirus, epidemic diarrhea, contagious gastroenteritis porcine rotavirus, epidemic encephalitis B, haemophilus parasuis, porcine circovirus and the like which are common in the piglets, the reserve swines and the sows can be effectively prevented.

The invention discloses a construction method for carriers for prokaryotic expression and eukaryotic expression of candidate gene of porcine rotavirus vaccines VP4 and VP7, comprising: cloning and prokaryotic expression of genes of porcine rotavirus VP4, construction and transfection of the recombinant vector of genes of rotavirus VP4 and VP7 and the expression carriers of insect cell sf9, and the construction of a recombinant vector of genes of rotavirus VP4 and VP7 and the expression vector of yeast cells GS115.

The invention discloses a porcine Deltacoronavirus (PDCoV) and swine transmissible gastroenteritis virus (TGEV) multiplex RT-PCR detection primer. The primer sequence of PDCoV is expressed as follows: upstream primer P1: 5'-ATGGCTACTGGCTGCGTTAC-3', downstream primer P2: 5'-GCGTTTCCTGGGCTGATT-3', and partial PDCoV gene segments are amplified by 383 bp. The primer sequence of TGEV is expressed as follows: upstream primer P3: 5'-CCCTCCAGCAAGGTTCAA-3', downstream primer P4: 5'-GCAACCCAGACAACTCCA-3', and partial TGEV gene segments are amplified by 229 bp. The detection limits of multiplex RT-PCR on PDCoV and TGEV are 4.05*101 copy per microliter and 5.47*102 copy per microliter respectively, and amplified results on porcine epidemic diarrhea virus, bocavirus, porcine reproductive and respiratory syndrome virus and porcine rotavirus are negative. Multiplex RT-PCR detection results of 57 clinical samples show that the positive rate of being infected with the two viruses at the same time is 1.75%, the PDCoV infection positive rate is 19.30%, and the TGEV infection positive rate is 1.75%.

The invention discloses a multiplex RT-PCR (reverse transcription-polymerase chain reaction) primer probe group for real-time fluorescent quantitative detection of four porcine epidemic diarrhea viruses. The multiplex RT-PCR primer probe group comprises an upstream primer, a downstream primer and a specific fluorescent probe of a porcine epidemic diarrhea virus M gene, an upstream primer, a downstream primer and a specific fluorescent probe of a swine transmissible gastroenteritis virus S gene; an upstream primer, a downstream primer and a specific fluorescent probe of a porcine rotavirus VP6 gene; and an upstream primer, a downstream primer and a specific fluorescent probe of a porcine D-type coronavirus M gene. The kit assembled on the basis of the primer probe group has the advantages of high sensitivity, high specificity, low pollution and real-time detection, and provides a reliable technology and product for early warning, early diagnosis and prevention and control monitoring of clinical diarrhea virus in a first-line pig farm.

An immune test paper for detecting group A rotaviruses comprises a sample pad, a bonding pad, a cellulose nitrate film, a water absorbing pad and a back lining, the sample pad, the bonding pad, the cellulose nitrate film and the water absorbing pad are boned on the back lining, two ends of the bonding pad have a lapping joint with the sample pad and the cellulose nitrate film respectively, one end of the cellulose nitrate film far from the binding pad has a lapping joint with the water absorbing pad, the cellulose nitrate film is provided with a detection line and a quality control line which are spaced, the bonding pad is provided with a group A porcine rotavirus monoclonal antibody A-fluorescent marker, the detection line is formed by a group A monoclonal porcine rotavirus antibody B, and the quality control line is formed by goat anti-mouse IgG. The invention also provides a making method of the immune test paper for detecting group A rotaviruses.

The invention discloses a porcine rotavirus strain and an inactivated vaccine prepared from the strain as well as application. The invention firstly discloses a separated porcine rotavirus zyh25 strain, of which the microbial preservation number is CGMCC No. 13792. The invention further discloses application of the porcine rotavirus zyh25 strain in the preparation of a vaccine for preventing porcine rotavirus diseases. Moreover, the invention discloses a vaccine composition for preventing the porcine rotavirus diseases, and the vaccine composition is prepared from an effective quantity of inactivated porcine rotavirus zyh25 strain used for prevention and treatment and an adjuvant which can be accepted in pharmacy. The porcine rotavirus zyh25 strain separated in the invention has good immunogenicity when serving as a vaccine strain, can generate a relatively high antibody level after used for immunizing an animal and can tolerate the attack of a virulent strain. The porcine rotavirus zyh25 strain can be used for serum-free full-suspended continuous production, and the prepared inactivated vaccine has the advantages of low production cost, small difference between product batches andthe like.

The invention discloses a porcine rotavirus strain as well as an inactivated vaccine prepared from the porcine rotavirus strain and application of the porcine rotavirus strain, and firstly discloses a separated porcine A-type rotavirus RVA/Pig-tc/CHN/SCMY-A3/2017/G9P [23] type strain (A3 strain for short), the microbial preservation number of the strain being V202132. The invention also discloses a preparation method of the porcine rotavirus inactivated vaccine, which comprises the following steps: mixing the virus liquid containing the A3 strain with the inactivating agent, inactivating the strain, and mixing the strain with the adjuvant to obtain the porcine rotavirus inactivated vaccine. The inactivated vaccine prepared from the separated porcine rotavirus can generate a higher antibody level and can resist the attack of virus strains.

The invention provides a porcine rotavirus VP6 and VP7 combined vaccine, which comprises vaccine antigen proteins and a vaccine adjuvant. The antigen proteins are a VP6 protein having an amino sequence represented by SEQ ID No.2 and a VP7 protein having an amino sequence represented by SEQ ID No.5. On the basis that a novel VP6 antigen gene is obtained through screening, the VP6 antigen protein isrecombinant expressed, and the expressed VP6 antigen protein and existing VP7 protein are used to produce a gene engineering subunit vaccine. The gene engineering subunit vaccine has a good immune effect on conventional porcine rotavirus and newly screened strains of rotavirus.

The invention relates to a composition containing probiotics metabolite. The composition is prepared from, by weight, 1-10 parts of probiotics metabolite, 2-3 parts of oligosaccharide and 3-8 parts ofpowder or liquid containing egg yolk antibody. Compared with the prior art, the composition containing the probiotics metabolite has the advantages that the waste discharge can be reduced, the wasteis turned into treasure, the bacteria reproduction can be selectively inhibited, and the composition is used for preventing or curing gastrointestinal infection caused by escherichia coli of piglets,salmonella, clostridium perfringens, porcine transmissible gastroenteritis virus, porcine epidemic diarrhea virus, porcine rotavirus and the like.