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A kind of pcr-rflp primer and method for distinguishing avian adenovirus type c and duck adenovirus type 3

A technology of PCR-RFLP and poultry adenovirus, which is applied in the field of PCR-RFLP primers for distinguishing poultry adenovirus type C and duck adenovirus type 3, and can solve unreported identification of poultry adenovirus type C and duck adenovirus type 3 molecules Biological methods, difficult to accurately determine, liver necrosis and other problems, to achieve high sensitivity, rapid identification, and strong specificity

Active Publication Date: 2022-05-17
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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AI Technical Summary

Problems solved by technology

Both avian adenovirus type C and duck adenovirus type 3 can infect ducks and cause liver necrosis, which is difficult to determine accurately clinically, and the molecular biology of identifying avian adenovirus type C and duck adenovirus type 3 has not yet been reported in this field method

Method used

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  • A kind of pcr-rflp primer and method for distinguishing avian adenovirus type c and duck adenovirus type 3
  • A kind of pcr-rflp primer and method for distinguishing avian adenovirus type c and duck adenovirus type 3
  • A kind of pcr-rflp primer and method for distinguishing avian adenovirus type c and duck adenovirus type 3

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Embodiment 1

[0033] 1. Materials

[0034] 1.1 Experimental reagents and consumables: 200 μL PCR tubes were purchased from Corning; viral nucleic acid extraction kit EasyPure Viral DNA / RNA Kit, bacterial genome extraction kit EasyPure Bacteria Genomic DNAKit were purchased from Beijing Quanshijin Biotechnology Co., Ltd.; Premix Taq TM (TaKaRa Taq TM Version 2.0plusdye) PCR Kit and QuickCut YM SpeI restriction endonuclease was purchased from Bao Biological Engineering (Dalian) Co., Ltd.

[0035] 1.2 Strains: Avian Adenovirus Type C (FJ1674 Strain), Duck Adenovirus Type 3 (AH06 Strain), Muscovy Duck Parvovirus (MDPV), Goose Parvovirus (GPV), Duck Plague Virus (DPV), Egg Drop Syndrome Virus (EDSV), duck-derived Escherichia coli (E.coli), duck-derived Riemerella anatipestifer (RA), and duck-derived Pasteurella multocida (PM) were all preserved by the Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences.

[0036] 2. Sequence analysis of hexon genes ...

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Abstract

The invention relates to a PCR-RFLP primer for distinguishing avian adenovirus type C and duck adenovirus type 3 and a method thereof. The primer sequence is: upstream primer HF: 5'-GAAAYTGGGGSCTGAAATAC-3'; downstream primer HR: 5 '‑TTGTAGTTYGTGGCCATCTG‑3', the primers designed by the present invention have strong specificity, can effectively amplify avian adenovirus type C and duck adenovirus type 3, but cannot amplify other common poultry pathogens; the present invention only A set of primers, the upstream primer HF and the downstream primer HR, combined with conventional XhoI restriction endonuclease digestion, can achieve rapid identification of avian adenovirus type C and duck adenovirus type 3, filling the relevant fields at home and abroad Research blank.

Description

technical field [0001] The invention belongs to the technical field of rapid diagnosis of veterinary infectious diseases, and in particular relates to a PCR-RFLP primer for distinguishing avian adenovirus type C and duck adenovirus type 3 and a method thereof. Background technique [0002] According to the classification of the International Committee on Taxonomy of Viruses (ICTV) (https: / / talk.ictvonline.org / taxonomy / ), Adenoviridae consists of 5 genera (Genus): Aviadenovirus, AT-rich gland Atadenovirus, Siadenovirus, Mastadenovirus and Ichtadenovirus. Adenovirus is a double-stranded DNA non-enveloped virus with icosahedral symmetry. The capsid is composed of 252 capsomers, of which 240 are hexons and 12 are penton bases. A penton base is combined with 1 to 2 fibers. The genome size of different adenoviruses varies, generally between 26-45kb. According to the start time of adenovirus DNA replication, its genome can be divided into early transcription genes (E) and late t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2521/301C12Q2565/125
Inventor 陈翠腾陈珍万春和施少华黄瑜程龙飞傅光华朱春华刘斌琼刘荣昌蔡国漳
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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