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One tube PCR type kit for discriminating I type duck hepatitis virus and new I type duck hepatitis virus

A duck hepatitis virus and kit technology, applied in the field of molecular biology, can solve the problems of inability to amplify, highly non-conservative strains, inability to amplify DHV-1A strains, etc., and achieve the effect of wide and economical scope.

Inactive Publication Date: 2013-08-21
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Wherein the upstream primer has designed the general primer DHF of DHV-1A and DHV-1C according to DHV-1A and DHV-1C highly conserved region: GGAGGTGGTGCTGAAATAT ( figure 1 ), the primers are highly conserved for DHV-1A and DHV-1C strains, so they are suitable for the amplification of DHV-1A and DHV-1C; The downstream specific primer of DHV-1A is DHV-1AR: CATGTGCCTGGACAGAT ( figure 2 ), this primer is highly conserved for DHV-1A, and highly variable for DHV-1C, therefore, this primer is only suitable for the amplification of DHV-1A, but cannot be amplified for DHV-1C strains; the downstream primer of DHV-1C is DHV-1CR: GTGTGGATCAAAGGGGT( image 3 ), the primer is completely conserved for the DHV-1C strain, but highly unconserved for the DHV-1A strain, and cannot amplify the DHV-1A strain

Method used

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  • One tube PCR type kit for discriminating I type duck hepatitis virus and new I type duck hepatitis virus
  • One tube PCR type kit for discriminating I type duck hepatitis virus and new I type duck hepatitis virus
  • One tube PCR type kit for discriminating I type duck hepatitis virus and new I type duck hepatitis virus

Examples

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Embodiment 1

[0030] Unless otherwise stated, the reagents used in the following examples are all commercially available reagents. Embodiment 1: One tube PCR formula distinguishes type 1 duck hepatitis virus (DHV-1A) and new type 1 duck hepatitis

[0031] Virus (DHV-1C)

[0032] 1. Handling of tested samples

[0033] Duck liver, spleen, kidney, cotton swab, duck / chicken embryo allantoic fluid and cell culture are used. The tested samples should be fresh (tested immediately after collection), or stored at -20°C. Add 3 times the volume of PBS to a 3g liver, spleen or kidney sample, grind it into a suspension, freeze and thaw 3 times, centrifuge at 1000g for 10 minutes, take the supernatant for RNA extraction; swab the cotton swab repeatedly, sample 1000g Centrifuge for 10 minutes, and take the supernatant for RNA extraction; duck / chicken embryo allantoic fluid and cell culture are directly used for RNA extraction.

[0034] Negative samples: select healthy 10-day-old chicken embryo or 13-da...

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Abstract

The invention discloses a one tube PCR type kit for discriminating duck hepatitis virus (DHV-1A) and new I type duck hepatitis virus (DHV-1C), and the kit comprises a primer, a reagent for treating sample to be tested, a PCR reagent and an instruction, wherein, the primer is SEQ ID Nos:1-3. The kit of the present invention is suitable for identifying DHV-1A virus and DHV-1C virus in liver, spleen and kidney tissue samples, cloaca cotton swab, duck or chicken embryo allantoic liquid and cell culture sample of duck.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a PCR-type test kit for identifying type I duck hepatitis virus and new type I duck hepatitis virus. The kit includes primers, reagents for processing samples to be tested, PCR reagents and instructions for use , wherein the primers are SEQ ID NOs: 1-3. The present invention also provides the use of the primers shown in SEQ ID NOs: 1-3 in preparing a PCR kit for identifying type I duck hepatitis virus DHV-1A and new type I duck hepatitis virus DHV-1C. Background technique [0002] Type I duck viral hepatitis is an acute and highly fatal infectious disease of ducklings caused by duck hepatitis virus serotype I (DHV-I) (Levine and Fabricant, 1950.). The traditional type I duck hepatitis virus (DHV-1A) belongs to the Picornaviridae family. Its viral nucleocapsid is a symmetrical icosahedron, has no envelope, has no hemagglutination, and has a diameter of 20-40 nm. In 1994,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 张云刘明陈玉环
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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