Primers for identifying attenuated vaccine strain and American strain of Chinese equine infectious anemia virus (EIAV)
A technology for equine infectious anemia and attenuated vaccine, applied in the field of specific primers, can solve the problems of affecting accuracy, inability to immunize horses with vaccines and identification of naturally infected horses with virulent strains, and inability to widely use serological detection methods, and achieve results. Simple judgment, short detection time, and the effect of meeting the detection requirements
Inactive Publication Date: 2011-01-05
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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Problems solved by technology
[0004] At present, the standard EIA detection methods recommended by the International Organization for Animal Health (OIE) are agar diffusion test and enzyme-linked immunosorbent assay, but these serological methods cannot distinguish between vaccine-immunized horses and naturally infected horses with virulent strains
In addition, these detection methods for antibodies in serum are affected by the growth and decline of antibodies in the body and affect the accuracy of detection
At the same time, due to the influence of maternal antibodies, serological detection methods cannot be widely used
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Embodiment 1
[0020] Differentiation of EIAV Vaccine Strain by LAMP Method DLV123 and American EIAV Wyoming , the design of the primers is the key. In the present invention, the PrimerExplorer V3 software provided by http: / / primerexplorer.jp / e / is used to screen each genome sequence to obtain 1000 LAMP primers. Through nucleic acid sequence alignment, find EIAV DLV123 with EIAV WyomingThere are fragments of sequence differences in gag, and then select the F3 / B3 primer pair containing the sequence of the difference region at the 3' end from 1000 primers, as the LAMP amplification primers of the respective strains ( figure 1 ,Table 1).
[0021] Table 1EIAV DLV123 and EIAV Wyoming strain LAMP primer
[0022]
[0023] LAMP reaction system: F3 / B3 / FIP / BIP (2.5μM) 2μl each, Beataine (5M) 4μl, MgSO 4 (100mM) 1.5μl, 10X ThermoPol Reaction Buffer 2.5μl, template DNA 2μl, Bst DNA Polymerase (8U) 1μl, add water to make up to 25μl. The reaction conditions were constant temperature at 65°C f...
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The invention discloses primers for identifying an attenuated vaccine strain and an American strain of a Chinese equine infectious anemia virus (EIAV). The primer for identifying the attenuated vaccine strain of the Chinese EIAV is composed of primer pair 1 and primer pair 2, wherein, the sequence of the primer pair 1 is shown as SEQ ID No.1 and SEQ ID No.2, and the sequence of the primer pair 2 is shown as SEQ ID No.3 and SEQ ID No.4; and the primer for identifying the American strain is composed of primer pair 3 and primer pair 4, wherein, the sequence of the primer pair 3 is shown as SEQ ID No.5 and SEQ ID No.6, and the sequence of the primer pair 4 is shown as SEQ ID No.7 and SEQ ID No.8. By adopting a loop-mediated isothermal amplification method established by the specific primer of the invention, the attenuated vaccine strain and the American strain of the Chinese EIAV can be accurately identified, thus the method has strong specificity and high sensitivity.
Description
technical field [0001] The invention relates to specific primers for identifying strains, in particular to specific primers for identifying Chinese equine infectious anemia virus attenuated vaccine strains and American strains, belonging to Chinese equine infectious anemia virus attenuated vaccine strains and American strains field of identification. Background technique [0002] Equine infectious anemia virus (EIAV) is a member of the lentivirus genus in the family Retroviridae and is the causative agent of equine infectious anemia (EIA). Most of the naturally infected horses were latent infection or after the initial acute onset, it turned to long-term latent infection. Since EIAV is the virus with the simplest genome among the seven lentiviruses including human AIDS virus, it is one of the ideal viruses for studying lentiviruses. [0003] In the 1970s, the attenuated EIA vaccine independently developed by China solved the problem of EIAV immune prevention and successful...
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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 林跃智李利周建华姜成刚王雪峰吕晓玲赵立平
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI