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A real-time fluorescent PCR kit for sheep arthritis-encephalitis virus provirus DNA

A real-time fluorescence and encephalitis virus technology, applied in the fields of molecular biology and nucleic acid detection, can solve the problems of cumbersome operation, difficult success, and long cycle

Inactive Publication Date: 2015-09-16
ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The virus isolation and culture method is not only long-term, cumbersome to operate, but also not easy to succeed

Method used

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  • A real-time fluorescent PCR kit for sheep arthritis-encephalitis virus provirus DNA
  • A real-time fluorescent PCR kit for sheep arthritis-encephalitis virus provirus DNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0019] Detection of Goat Arthritis-Encephalitis Virus Proviral DNA in Goat Peripheral Blood Lymphocytes

[0020] 1. Isolation of Peripheral Blood Mononuclear Cells (PBMCs)

[0021] (1) Collect 2mL goat whole blood, add 2mL anticoagulant (EDTA) to it;

[0022] (2) Add 4.0mL of lymphocyte separation medium to the centrifuge tube, slowly add 2mL of diluted anticoagulant blood to the surface of the separation medium, and centrifuge horizontally at 2000rpm for 30min to separate the liquid;

[0023] (3) Use a capillary glass tube to absorb the middle cloud layer cells, namely peripheral blood mononuclear cells, wash the collected peripheral blood mononuclear cells with 10mL PBS once, collect the cells by centrifugation at 1500rpm, repeat once, the peripheral blood mononuclear cells It can be used for the extraction of genomic DNA from samples.

[0024] 2. Extract the DNA of the sample to be tested

[0025] Use commercially available blood / cell / tissue genomic DNA extraction kits f...

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Abstract

The invention discloses a real-time fluorescence PCR (polymerase chain reaction) kit of caprine arthritis-encephalitis virus and a detection method. The kit is high in detection sensitivity, good in specificity, simple and convenient to operate, fast and capable of avoiding false positive of the conventional PCR method.

Description

technical field [0001] The invention belongs to the fields of molecular biology and nucleic acid detection, and particularly relates to a real-time fluorescent PCR kit and a detection method for detecting goat arthritis-encephalitis virus proviral DNA in goat peripheral blood lymphocytes. Background technique [0002] Caprine arthritis-encephalitis (CAE) is a chronic infectious disease of goats caused by the retroviridae lentivirus genus caprine arthritis encephalitis virus (CAEV). It is characterized by arthritis, sclerosing mastitis, and interstitial pneumonia. The disease is widely distributed in the world, and the seropositive detection rate in Canada, the United States, Australia, Switzerland, France and other countries can reach as high as 65% to 77%. The disease was brought into our country when we imported breeding goats in the 1980s. Therefore, strict entry quarantine of imported breeding goats is one of the fundamental measures to prevent the disease from being in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor 王建华赵祥平陈本龙王玉玲王乃福
Owner ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU