Peste des petits ruminants virus (PPRV) RT-LAMP kit

A technology of RT-LAMP and Peste des petits ruminants, which is applied in the biological field to achieve the effects of simple operation, easy reaction results, and easy popularization and application on a large scale

Active Publication Date: 2009-12-30
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no application of this method in the detection of Peste des petits ruminants virus.

Method used

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  • Peste des petits ruminants virus (PPRV) RT-LAMP kit
  • Peste des petits ruminants virus (PPRV) RT-LAMP kit
  • Peste des petits ruminants virus (PPRV) RT-LAMP kit

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Embodiment 1

[0063] 1. Design of primers

[0064] Using Primer 4.0 ( http: / / primerexlorer.jp Eiken Chemical Co.Ltd, Japan), designed four primers for the conserved region (GeneBank, gi: 397561, conserved region 1-700bp) of the PPRV N gene, including two internal primers (FIP and BIP) and two external primers ( F3 and B3)

[0065] 2. RT-LAMP

[0066] 1. Extraction of viral RNA:

[0067] Take PPR virus vaccine strain Nigeria 75 / 1 (product of Xinjiang Tiankang Biological Products Factory: PPR attenuated vaccine). Use TaKaRa Mini BEST Viral RNA / DNAExtraction Kit Ver.3.0 Kit (TaKaRa Company), extract viral total RNA from 250 μL LPPRV cell culture medium according to the instructions, and finally dissolve in 30 μL elution buffer (Elution Buffer), add 1 μL RNase Inhibitor (TaKaRa Company) (20U), stored in -80°C refrigerator for later use.

[0068] 2. Establishment of RT-LAMP detection system

[0069] Referring to Notomi et al. (Notomi, T., Okayama, H., Masubuchi, H., et al. Loop-mediated i...

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Abstract

The invention relates to the biotechnology field, in particular to an RT-LAMP kit for detecting peste des petits ruminants virus (PPRV). The kit comprises a detecting system composed of LAMP reaction liquid of four LAMP primers. The kit can quickly and sensitively detect PPRV, has simple operation, low cost, easy observation of reaction results and favourable specificity, is suitable for export quarantine, food sanitation and on-site detection of livestock feedlots, and is easy for popularizing and applying in large range.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an RT-LAMP kit for detecting Peste des Petits Ruminants virus. Background technique [0002] Peste des petits ruminants virus (PPRV) belongs to the genus Morbillivirus in the Paramyxoviridae family. The peste des petits ruminants virus (PPRV) caused by it is listed as an animal infectious disease that must be reported by the World Organization for Animal Health. It is a severe and contact infection. The disease mainly infects small ruminants, especially goats, which are highly susceptible, with a morbidity rate of up to 100% in susceptible groups, and a fatality rate of 100% in severe outbreaks. [0003] The disease was first discovered in Côte d'Ivoire in West Africa in 1942. In recent years, the disease has been spreading and the scale is getting bigger and bigger. Recently, large-scale outbreaks of Peste des petits ruminants have occurred in neighboring countries of my country (L...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
Inventor 李刚李伟赫尔曼·昂格尔
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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