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VP7 gene-based bovine rotavirus qPCR detecting kit

A detection kit, bovine rotavirus technology, applied in the biological field, can solve problems such as no bovine rotavirus diagnosis

Inactive Publication Date: 2014-11-19
魏锁成 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no specific kit for the diagnosis of bovine rotavirus

Method used

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  • VP7 gene-based bovine rotavirus qPCR detecting kit
  • VP7 gene-based bovine rotavirus qPCR detecting kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0014] Sample collection and processing

[0015] (1) Tissue sample processing. Each tissue was weighed about 1g sample from three different positions, cut and mixed with surgical scissors, then took 0.05g and ground it in a grinder, added 1.5 mL normal saline to continue grinding, and transferred to 1.5 mL after homogenization. Centrifuge at 8,000 rpm for 2 min, and take 100 μL of the supernatant into a 1.5 mL sterilized centrifuge tube.

[0016] (2) Whole blood sample processing. After hemagglutination, take 100 μL of serum and put it in a 1.5 mL sterilized centrifuge tube.

[0017] (3) Positive control treatment. Take 100 μL of the positive control and place it in a 1.5 mL sterilized centrifuge tube.

[0018] (4) Negative control treatment. Take 100 μL of the negative control and put it in a 1.5mL sterilized centrifuge tube.

[0019] RNA extraction

[0020] Take the processed sample, negative control and positive control, add 600 μL of lysate respectively, mix well by...

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PUM

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Abstract

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.

Description

technical field [0001] The invention relates to a bovine rotavirus qPCR detection kit based on VP7 gene, belonging to the field of biotechnology. [0002] Background technique [0003] Rotavirus (rotavirus, RV) is an important cause of severe dehydrating diarrhea in infants and young animals worldwide. Rotavirus gastroenteritis is a widespread zoonotic disease, especially an acute intestinal infectious disease of young animals and infants. It is characterized by diarrhea and dehydration, and the case fatality rate can reach 50%. The genome of rotavirus consists of 11 discontinuous dsRNA fragments, encoding 6 structural proteins (VP1-VP4, VP6, VP7) and 5 non-structural proteins (NSP1-NSP5); VP7 and VP4 constitute the outer coat Shell, VP7 has serotype specificity, is related to pathogenicity, and can stimulate the body to produce protective antibodies. VP7 is an N-linked oligomannose glycoprotein, which is an important glycoprotein of the outer membrane of the virus and th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/701C12Q1/6851C12Q1/70C12Q2531/113C12Q2561/113
Inventor 魏锁成郭慧琳车团结
Owner 魏锁成
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