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Primer pair, primer group and kit for detecting feline leukosis viruses and application thereof

A leukemia virus and kit technology, applied in the field of molecular biology, can solve problems such as misdiagnosis, non-existence, interference, etc., and achieve the effect of reducing the misdiagnosis rate, reducing interference, and efficient detection

Inactive Publication Date: 2018-10-19
联宠科技(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Feline endogenous leukemia virus has great interference on molecular biology detection of FeLV infection, and it is easy to misdiagnose
So far, there is no method or kit that can distinguish exogenous and endogenous FeLV in my country

Method used

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  • Primer pair, primer group and kit for detecting feline leukosis viruses and application thereof
  • Primer pair, primer group and kit for detecting feline leukosis viruses and application thereof
  • Primer pair, primer group and kit for detecting feline leukosis viruses and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0044] 1 Design of PCR primers for feline leukemia virus (FeLV)

[0045] 1.1 Design of primers

[0046] The present invention finds that the endogenous and exogenous sequences of the feline leukemia virus have different base sequences on the LTR, and both the endogenous and exogenous FeLV contain gp70 gene sequences encoding glycoproteins. By referring to and comparing all FeLV LTR and gp70 gene sequences in GenBank, the exogenous FeLV-specific nucleotide sequence and gp70 conserved sequence were respectively selected as the amplification region, and two pairs of feline exogenous and endogenous feline detection were designed. PCR primers derived from leukemia virus;

[0047] A pair of PCR primers for detecting exogenous FeLV, the sequences of the PCR upstream and downstream primers FeLV-1F / R are shown in sequence as SEQ ID NO: 1 to SEQ ID NO: 2:

[0048] Upstream primer FeLV-1F: 5'-GTTCGACCTTCCGCCTCAT-3' (SEQ ID NO: 1);

[0049] Downstream primer FeLV-1R: 5'-ACGAGTCAGATGCAA...

experiment example 1

[0068] Validation of Feline Leukemia Virus PCR Primers.

[0069] The experiment was carried out with reference to the steps in the examples, wherein No. 1 was a sample containing both feline exogenous and endogenous leukemia virus, No. 2 was a sample containing only feline exogenous leukemia virus, and No. 3 was a sample containing only cat endogenous leukemia virus. The sample of leukemia virus derived from the sample, No. 4 is the blank control.

[0070] The result is as figure 1 shown. The PCR product of sample No. 1 had obvious bright bands at 177bp and 892bp, and the negative control (No. 4) had no bands, indicating that sample No. 1 contained feline exogenous and endogenous leukemia virus; the PCR product of sample No. 2 was in There is an obvious bright band at 177bp, and there is no band in the negative control, indicating that sample No. 2 only contains feline exogenous leukemia virus and does not contain feline endogenous leukemia virus; the PCR product of sample N...

experiment example 2

[0072] Validation of the specificity of PCR primers for feline leukemia virus.

[0073] Carry out the experiment with the steps in the reference examples, with the genomic DNA or cDNA of feline leukemia virus, feline AIDS virus, feline herpes virus, feline calicivirus, feline distemper virus and feline coronavirus as template, carry out PCR with the primers described in this application detection.

[0074] The result is as figure 2 As shown, only No. 1 cat endogenous and exogenous leukemia virus-positive samples had bright and clear bands at 177bp and 892bp, and the other groups of samples had no bands. The results show that the feline leukemia virus PCR primers designed by the present invention have high specificity, only feline endogenous and exogenous leukemia virus are positive, while feline AIDS virus, feline herpes virus, feline calicivirus, feline distemper virus and cat All tested negative for coronavirus.

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Abstract

The invention relates to the field of molecular biology and particularly relates to a primer pair, a primer group and a kit for detecting feline leukosis viruses and application thereof. The primer pair is designed by taking 3'LTR nucleotide sequence conserved by the exogenous FeLVs as an amplification region. The primer pair has the advantages of high sensitivity, excellent specificity and convenience in detection; the primer group comprises the primer pair and a control primer pair for detecting the cat endogenous leukemia virus; the primer group is capable of rapidly, conveniently and efficiently detecting exogenous and endogenous FeLVs and is excellent in specificity and high in sensitivity. The primer pair, the primer group and the kit have guiding significance to development of clinical detection and epidemiological investigation, and can provide certain guarantee for safe development of the pet market.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a primer pair, primer set, kit and application for detecting feline leukemia virus. Background technique [0002] Feline leukemia virus (FeLV) is a retrovirus belonging to the Oncoviridae subfamily, which can cause feline leukemia. Persistent infection can cause severe immunosuppression, severe nonregenerative anemia, and tumors. FeLV has four subtypes, FeLV-A, FeLV-B, FeLV-C and FeLV-T. Type A is present in almost all FeLV-infected cats and can cause severe immunosuppression. Type B is present in approximately 50% of FeLV-infected cats and can cause tumors. Type C is present in only about 1% of FeLV-infected cats and can cause severe anemia. If no immune response occurs after infecting the body, the virus may cause persistent viremia (40%) to become a carrier of the virus. Viremia usually occurs 2 to 4 weeks after FeLV infection, and about 50% of cats with persistent viremia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/702C12Q2600/166C12Q2545/113Y02A50/30
Inventor 安娜张伟伟刘潇然蒋勤军
Owner 联宠科技(北京)有限公司
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