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Pathogenic detection kit for lentivirus of small ruminant

A technology for ruminants and kits, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., which can solve the problems of quarantine failure and timely diagnosis of virus infection

Pending Publication Date: 2017-12-08
新疆畜牧科学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the virus infection can only be stimulated to produce serological antibodies after 3 months, the virus infection cannot be diagnosed in time, which may easily lead to quarantine failure

Method used

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  • Pathogenic detection kit for lentivirus of small ruminant
  • Pathogenic detection kit for lentivirus of small ruminant
  • Pathogenic detection kit for lentivirus of small ruminant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1, primer sequence: two primers were designed according to the DNA sequence of the disclosed small ruminant lentivirus in the GeneBanK of NCBI, P1 is the upstream primer; P2 is the downstream primer, and carried out relevant parameter analysis, see Table 1, Entrust a biotechnology company to carry out the synthesis.

[0020] Table 1 Sequence and related parameters of the two sequences

[0021]

[0022] 2. Detection Kit

[0023] 2.1 Standards

[0024] Negative control standard: sterile pure water without nucleotide contamination, pH6.8-7.2.

[0025] Positive control standard: Use artificially synthesized DNA fragments at a concentration of 0.5 μg / mL.

[0026] Table 2 Positive Control Standard Nucleotide Sequence

[0027]

[0028] 2.2 Reagents

[0029] 1mL 2x PCR Reagent (KT207-01) and 0.3mL D2000DNA Marker (MD114), produced by Tiangen Biochemical Technology (Beijing) Co., Ltd.; 5mL sterile pure water without nucleotide contamination, pH6.8-7.2; Prim...

Embodiment 2

[0049] Embodiment 2, use PCR method

[0050] After caprine arthritis-encephalitis virus (CAEV) infects goats, the viral genetic material RNA is integrated into the host cell genome. By extracting the total DNA of white blood cells in goat blood, the established PCR method is used to detect the infection of goats with CAEV.

[0051] 1.1 Materials

[0052] Four copies of goat anticoagulant blood were screened by IDEXX CAEV / MVV Total Antibody Screening Kit (product number: CVT1135T), and the antibody was positive.

[0053] 1.2 Test method

[0054] The detection steps are as follows:

[0055] (1) Extraction of blood genome

[0056] Use the "Blood Genomic DNA Extraction Kit (0.1-1ml) (DP348)" produced by Tiangen Biochemical Technology (Beijing) Co., Ltd. for whole blood genome extraction;

[0057] (2) Carry out according to the PCR method in the above-mentioned technical scheme.

[0058] 2. Results

[0059] Screened by IDEXX CAEV / MVV Total Antibody Screening Kit, the serum anti...

Embodiment 3

[0061] Embodiment 3, use PCR method to detect sheep to be infected with MVV

[0062] After sheep are infected by Medy-Visna virus (MVV), the viral genetic material RNA is integrated into the host cell genome. By extracting the total DNA of white blood cells in sheep blood, the established PCR method is used to detect the infection of sheep with MVV.

[0063] 1.1 Materials

[0064] Sheep anticoagulant was screened by IDEXX CAEV / MVV total antibody screening kit (product number: CVT1135T), and the antibody was positive.

[0065] 1.2 Test method

[0066] The detection steps are as follows:

[0067] (1) Extraction of blood genome

[0068] Use the "Blood Genomic DNA Extraction Kit (0.1-1ml) (DP348)" produced by Tiangen Biochemical Technology (Beijing) Co., Ltd. for whole blood genome extraction;

[0069] (2) Carry out according to the PCR method in the above-mentioned technical scheme.

[0070] 2. Results

[0071] Screened by IDEXX CAEV / MVV Total Antibody Screening Kit, 9 samp...

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Abstract

The invention belongs to the technical field of molecular biological detection and in particular relates to a pathogenic detection kit for a lentivirus of a small ruminant. The kit comprises (1) a genetic marker for detecting the lentivirus of the small ruminant, wherein the genetic marker has a sequence as shown in SEQ ID NO.3 or other sequences, the sequence consistency of which is greater than 80%; (2) a specific primer pair for amplifying the genetic marker, the specific primer pair comprising an upstream primer sequence as shown in SEQ ID NO.1 and a downstream primer sequence as shown in SEQ ID NO.2 or other sequences, the consistency of any continuous eight nucleotide base sequences of which is greater than 80%, among the upstream and downstream primer sequences; and (3) a kit for detecting the lentivirus of the small ruminant, wherein the kit comprises the specific primer pair in (2).

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, and in particular relates to an etiological detection kit of small ruminant lentivirus. Background technique [0002] Small ruminant lentiviruses, including caprine arthritis-encephalitis (CAEV) and medi-Visna virus (MVV), can cause a chronic viral infectious disease in goats (CAEV) or sheep (MVV). Caprine Arthritis-Encephalitis Virus (CAEV), the main disease in adult goats is characterized by severe wrist arthritis ("big knees"), even if appetite is not affected, CAEV-infected animals become very emaciated, produce There is also very little milk. Serological surveys have shown that CAEV is widely distributed in goat populations on all continents; after infection with Maedi-Visna virus (MVV), the main symptoms of sheep include chronic progressive arthritis, pneumonia, mastitis and encephalomyelitis. The causative agent of this disease is a lentivirus, which belongs to the su...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/702C12Q2600/166
Inventor 王登峰吴建勇杨义琴古努尔·吐尔逊李建军刘志强杨学云王文奇刘艳丰马晓菁叶锋金映红刘丽娅班万里蒋晓梅
Owner 新疆畜牧科学院
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