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

CN110527748BActive Publication Date: 2022-05-17INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
5 Cites 0 Cited by

Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Publication Date
2022-05-17

Smart Images

  • Figure 1
    Figure 1
  • Figure 2
    Figure 2
  • Figure 3
    Figure 3
Patent Text Reader

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.
Need to check novelty before this filing date? Find Prior Art

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...

Examples

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 ...