Porcine reproductive and respiratory syndrome and porcine Japanese B encephalitis dual one-step RT-PCR (Reverse Transcription-Polymerase Chain Reaction) diagnosis kit

A RT-PCR, respiratory syndrome technology, applied in biochemical equipment and methods, microorganism-based methods, and microbial assay/inspection, etc., can solve problems such as serious reproductive barriers, low sensitivity, and complicated operations.

Active Publication Date: 2013-07-24
GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] With the continuous development of large-scale pig farming, the reproductive barriers caused by PRRSV and JEV are more serious, and mixed infections often occur, causing huge economic l

Method used

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  • Porcine reproductive and respiratory syndrome and porcine Japanese B encephalitis dual one-step RT-PCR (Reverse Transcription-Polymerase Chain Reaction) diagnosis kit
  • Porcine reproductive and respiratory syndrome and porcine Japanese B encephalitis dual one-step RT-PCR (Reverse Transcription-Polymerase Chain Reaction) diagnosis kit
  • Porcine reproductive and respiratory syndrome and porcine Japanese B encephalitis dual one-step RT-PCR (Reverse Transcription-Polymerase Chain Reaction) diagnosis kit

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Embodiment Construction

[0062] Embodiment of the present invention: Porcine reproductive and respiratory syndrome and porcine Japanese encephalitis dual one-step RT-PCR diagnostic kit, including DL2000, RT-PCR one-step enzyme, buffer, RNase Free dH2O, mixed primers, positive control and negative control; mixed primers include PRRSV upstream primer, PRRSV downstream primer, JEV upstream primer and JEV downstream primer; the sequence of PRRSV upstream primer is: 5'-CGCCCAACAAAACCAGTCCAGA-3', the sequence of PRRSV downstream primer is: 5'-CAGTATATTGCTCGGCAAACT- 3'; the sequence of the JEV upstream primer is: 5'-TCCTCGTCATAGGGATTTGCT-3', the sequence of the JEV downstream primer is: 5'-AGGCCCAAGGCGTGAGGGCGG-3'; the negative control is ultrapure water; the positive control is standard porcine reproductive and respiratory syndrome And pig Japanese encephalitis RNA RT-PCR product; Described RT-PCR one-step method enzyme is made up of the MgCl of 50% glycerol, 3 mmol / μL, the Tris-HCl of 25mmol / μL and 0.5 μ L ...

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Abstract

The invention discloses a porcine reproductive and respiratory syndrome and porcine Japanese B encephalitis dual one-step RT-PCR (Reverse Transcription-Polymerase Chain Reaction) diagnosis kit which comprises 100 micro-milliliters of DL2000, 20 micro-milliliters of RT=PCR one-step enzyme, 250 micro-milliliters of enzyme buffer solutions, 300 micro-milliliters of RNase Free dH2O, 80 micro-milliliters of mixed primers ( respectively with the concentration of 10 micromoles/liter), 20 micro-milliliters of positive controls and 20 micro-milliliters of negative controls. According to the invention, two pairs of primers are designed according to the sequences of a PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) OFR7 N gene and a JEV (Japanese Encephalitis Virus) NS1 gene which are contained in a GenBank; and the dual one-step RT-PCR diagnosis kit for detecting a porcine reproductive and respiratory syndrome virus and a porcine Japanese B encephalitis virus is successfully developed through the optimization of reaction conditions, and the dual one-step RT-PCR diagnosis kit has sensibility and specificity on the two viruses. The dual one-step RT-PCR diagnosis kit disclosed by the invention can be used for clinically detecting the porcine reproductive and respiratory syndrome and the porcine Japanese B encephalitis.

Description

technical field [0001] The invention relates to a kit, in particular to a dual one-step RT-PCR diagnostic kit for porcine reproductive and respiratory syndrome and porcine Japanese encephalitis. Background technique [0002] Porcine reproductive and respiratory syndrome is an infectious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which causes reproductive disorders in pregnant sows and respiratory symptoms in piglets, commonly known as porcine blue ear disease. Japanese encephalitis virus (JEV), also known as Japanese encephalitis virus, is a zoonotic viral infectious disease that causes reproductive disorders in sows and central nervous system diseases in humans. The disease not only affects animal husbandry, especially pig raising industry and seriously threaten human health [3-5]. [0003] With the continuous development of large-scale pig farming, the reproductive barriers caused by PRRSV and JEV are more serious, and mixed infections ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 余波冉懋韬谭诗文史开志
Owner GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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