37 results about "Group A rotaviruses" patented technology

The invention discloses a single standard product-based quardruple four-color fluorogenic quantitative PCR (Polymerase Chain Reaction) technology. The technology is capable of detecting group A rotavirus (HRVA), norovirus group I (NVGI), norovirus group II (NVG II) and human astrovirus (HAstV) RNA in various treated specimen through one-step quardruple PCR amplification and quantifying virus load. The technology is characterized in that single standard product is adopted as positive control and quantitative standard in the quardruple four-color fluorogenic quantitative PCR detection for four viruses, thereby breaking through the deficiency that the traditional multiple fluorogenic quantitative PCR needs a plurality of standard products, improving the experiment detection efficiency, reducing the pollution probability and improving the preparation and production efficiency of the kit. The method and kit can be used for fast high-throughput test and epidemic monitoring in laboratories of HRVA, NVG I, NVG II and HAstV which can cause diarrhea syndromes. The single standard product-based four-color florogenic quantitative PCR method and the kit belong to the biotechnology field.

The invention discloses a kit and detection method for accurate quantitative detection of group A rotaviruses, and particularly relates to a set of primers and probes having nucleotide sequences shown in sequence tables of SEQ ID No.1 to SEQ ID No.3 used for detecting the group A rotaviruses, and the kit containing the primers and probes and the detection method for the group A rotaviruses. The detection method for the accurate quantitative detection of the group A rotaviruses by using a droplet digital PCR (Polymerase Chain Reaction) technology provided by the invention has higher sensitivity, accuracy and repeatability, and is easily standardized.

The invention discloses a composition for detecting group A rotavirus, belonging to the technical field of biology, comprising the following primers: 5'-GGCTTTAAAAGAGAGAATTTCC-3' and 5'-AAYTGYGGTATATTCAATACCATACA-3', also comprising the following probes: 5'-fluorescent reporter dye AARGAGCTAWCCGYTAGCCAGACGG-fluorescence quencher-3'. The invention further discloses a kit containing the primers andprobes and a method for detecting group A rotavirus. When using the composition, the kit and the/or method to detect group A rotavirus, a genome equivalent/ reaction system with the range of 10<8>-10<1> copies is detected, the detection is rapid and simple, and the whole detection process can be completed within 3 hours with high specificity and high accuracy.

The invention relates to an isothermal nucleic acid amplification system with high specificity. The isothermal nucleic acid amplification system with high specificity comprises overall-length Bst DNApolymerase and also comprises embedded fluorescent dye. The invention also relates to a kit containing the isothermal nucleic acid amplification system as well as the application of the isothermal nucleic acid amplification system to childhood diarrhea pathogen detection. In the application, amplification primers comprise primer groups which are as shown in SEQ ID NO.1 to SEQ ID NO.4, SEQ ID NO.5to SEQ ID NO.8 and SEQ ID NO.9 to SEQ ID NO.12 and are correspondingly used for detecting Group A rotaviruses, astroviruses and adenoviruses. The overall-length Bst DNA polymerase has 5'-3'-duplex-specific excision enzyme activity and polymerization activity, the characteristic is applied to the Exo-NAT method of the invention, the embedded fluorescent dye is added into the reaction system and melting curve analysis is cooperated, so that high-specificity and high-sensitivity multiple nucleic acid detection can be realized; and the isothermal nucleic acid amplification system is successfully applied to accurate detection of childhood diarrhea pathogens, and has an important clinical significance and a wide application prospect.

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 method for simultaneously detecting three groups of rotaviruses A, B and C, and belongs to the field of biological detection. A pair of specific primers and a specific probewith different fluorescent labels are designed for target genes of group A rotavirus, group B rotavirus and group C rotavirus, a pair of universal primers is further designed, and during PCR amplification, the specific primers with universal primer tags are used for enrichment and amplification; and then universal primer tags are used for exponential amplification, and fluorescence signals of exponential amplification products are collected for detection. The method can simultaneously detect the group A rotavirus, the group B rotavirus, and the group C rotavirus. The method is high in specificity, high in sensitivity and short in detection time, and can be used for early rapid diagnosis of rotavirus infection and epidemiological study of the rotavirus.

The invention discloses a group-A rotavirus rapid real-time fluorescent RT-PCR (reverse transcription-polymerase chain reaction) detection kit which comprises an amplification primer, a forward primer 5'-gttgatgctcaagatggagt-3', a reverse primer 5'-acttcattgtaatcatattgaatacc-3' and a specific fluorescent probe 5'-X-cagcaacaactgcagcttcaaaagaagtgt-Y-3'. The kit can meet the requirement of timeliness for group-A rotavirus detection, group-A rotaviruses are rapidly and quantitatively detected in real time according to one-step fluorescent RT-PCR technology by the design of the primers, the probe and reaction conditions in a reaction system and heat-resisting DNA (deoxyribonucleic acid) rapid polymerase, and sensitivity can reach 10<2> PFU/ml.

The invention belongs to the field of food detection, and particularly relates to a group A rotavirus chromatography test paper strip based on low-noise excitation fluorescent label as well as a preparation method and application of the chromatography test paper strip. According to the invention, quantum dots are adopted as the label, the immunochromatography technology is adopted, so that the chromatography test paper strip targeting to group A rotavirus is prepared, during detection, a special portable low-noise excitation fluorescent scanner is adopted for scanning a quality control line and a detection line, and qualitative and quantitative determination of a sample is realized through a fluorescence intensity detection value of the detection line. The test paper strip is used for immediate detection of group A rotavirus in food and a pathological sample, and has the advantages of rapidness, high sensitivity, qualitative, accurate and quantitative detection, and portability.

The invention provides a new method for rapid detection of G genotype of group A rotavirus by VP7 gene, comprising the steps of: 1) designing six nucleotide sequences capable of specifically amplifying different G genotypes of group A rotavirus as the primers for detecting the different G genotypes; 2) extracting virus nucleic acid from a sample and conducting one-time RT-PCR (reverse transcription-polymerase chain reaction) amplification with the six primers, and performing an electrophoresis detection to a product of PCR with 2% agarose gel, dyeing the product with ethidium bromide, and taking a picture by means of a gel imaging system; 3) judging the G genotype of group A rotavirus: when the size of the PCR product is respectively 332bp, 600bp, 98bp, 471bp and 248bp, the sample respectively contains G1, G2, G3, G4 and G9 rotaviruses, thus establishing a criteria for judging the G genotype of group A rotavirus.

The invention discloses a traditional Chinese medicine for treating infant group A rotavirus infectious diarrhea. The traditional Chinese medicine is characterized by comprising 10-15g of kudzu vine roots, 10-15g of scutellaria baicalensis, 10-15g of coptis chinensis, 5-10g of agastache rugosus, 5-10g of talc, 5-10g of radix saposhnikoviae, 5-10g of rhizoma atractylodis, 4-6g of liquorice, 8-12g of medicated leaven, 8-12g of roasted malt, 8-12g of hawthorn and 8-12g of poria cocos. The components are decocted twice till a decoction of 100mL is obtained, and 20-30mL of the decoction is kept being 37 DEG C for enema. The traditional Chinese medicine for treating infant group A rotavirus infectious diarrhea, which is disclosed by the invention, is capable of relieving stagnation and activating spleen, rhizoma atractylodis has the effects of drying humidity and tonifying spleens, exogenous pathogen can be removed, intestines can be cleared, humidity can be removed, and together with a retention enema administration way of the traditional Chinese medicine, the treatment effects of the traditional Chinese medicine can be brought into play, the phenomenon that spleens and stomachs are injured as the decoction enters the stomachs can be avoided, affected parts can be reached directly, and the purpose of treatment can be achieved.

The invention provides an oligonucleotide chip for synchronous detection of four porcine diarrhea viruses. The oligonucleotide chip comprises a solid phase carrier and oligonucleotide probes fixed onthe solid phase carrier, wherein the oligonucleotide probes comprise a probe shown in SEQ ID No. 1-2, a probe shown in SEQ ID No. 3-4, a probe shown in SEQ ID No. 5-6, and a probe shown in SEQ ID No.7-8; the probes are respectively used for detecting porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV), porcine group A rotavirus (GAR) and porcine delta coronavirus (PDCoV). The invention also provides a kit and a method for the synchronous detection of the four porcine diarrhea viruses. The chip and the method can realize the high flux co-detection of thefour porcine diarrhea viruses, i.e., PEDV, TGEV, GAR and PDCoV, and can be applied to mass detection of clinical diseases, thus having a wide application prospect.

The invention relates to the field of viruses, in particular to separation, culture and identification of a group A rotavirus G9P[8] strain and application of the group A rotavirus G9P[8] strain as a vaccine. Specifically, the invention relates to a group A rotavirus seed LSR-8 strain, and relates to a group A rotavirus seed LSR-8 strain; the total amino acid sequences of the group A rotavirus seed LSR-8 strain are shown as SEQ ID No.1 to SEQ ID No.11; and the total nucleic acid sequences of the group A rotavirus seed LSR-8 strain are shown as SEQ ID No.12 to SEQ ID No.22. The invention further relates to application of the LSR-8 strain in production of inactivated human rotavirus vaccines, human rotavirus attenuated live vaccines and combined vaccines; and the LSR-8 strain can be used for preventing, relieving or treating rotavirus infection and diseases caused by rotavirus infection, such as rotavirus gastroenteritis, diarrhea and the like.