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Chicken infectious anemia virus detection method based on CRISPR/Cas12a system

A technology for chicken infectious anemia and detection method, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, and microorganism determination/inspection, etc. problem, to achieve high sensitivity and specificity

Pending Publication Date: 2022-07-22
ANHUI AGRICULTURAL UNIVERSITY +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to its relatively slow onset, no obvious symptoms, long duration of onset, and low mortality, it affects the growth and production performance of poultry

Method used

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  • Chicken infectious anemia virus detection method based on CRISPR/Cas12a system
  • Chicken infectious anemia virus detection method based on CRISPR/Cas12a system
  • Chicken infectious anemia virus detection method based on CRISPR/Cas12a system

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Embodiment Construction

[0060] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, but not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0061] The present invention provides a technical solution: a method for detecting chicken infectious anemia virus based on CRISPR / Cas12a system, comprising the following steps:

[0062] S1. Treatment of tissue samples: about 1 g of each tissue sample will be weighed, added with 1 mL of PBS, put into a grinding bowl and ground into a homogenate, transferred to a sterile centrifuge tube, and centrifuged at 8500 rpm / min for 3 min in a ce...

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Abstract

The invention relates to the technical field of gene engineering, and discloses a chicken infectious anemia virus detection method based on a CRISPR / Cas12a system, which comprises the following steps: S1, treatment of tissue samples: weighing about 1g of each tissue sample, adding 1mL of PBS (Phosphate Buffer Solution), putting into a grinding bowl, grinding into homogenate, transferring into a sterile centrifugal tube, and centrifuging in a centrifugal machine at 8500rpm / min for 3 minutes; s2, virus nucleic acid extraction: extracting virus RNA by using a virus genome DNA / RNA extraction kit (centrifugal column type) of a certain biochemical technology Co., Ltd; a CRISPR / Cas12a system is combined with an ERA technology, the rapid molecular detection method for the chicken infectious anemia virus is established, the method has the advantages of visualization, high sensitivity, strong specificity, no dependence on expensive instruments and equipment and the like, the whole detection process only needs about 1 h, a new means is provided for rapid diagnosis of the chicken infectious anemia virus, and the method is suitable for large-scale popularization and application. And the method is of great significance to monitoring of immunosuppressive viruses.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for detecting chicken infectious anemia virus based on a CRISPR / Cas12a system. Background technique [0002] Infectious chicken anemia virus (CIAV) is a disease caused by a variety of infectious and non-infectious factors, which are caused by a single pathogen or a combination of different pathogenic factors, resulting in damage to the immune system of poultry and a significant decline in immune function. Because its onset is relatively slow, the symptoms are not obvious, the onset duration is long, and the mortality rate is not high, but it affects the growth and production performance of poultry. In recent years, the phenomenon of poultry infection with immunosuppressive virus has generally appeared in many parts of the world, which has caused great harm to chicken flocks and poultry industry. [0003] Based on this, this study combines the CRISPR / Cas12a s...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/701C12Q1/686C12Q2600/166C12Q2521/327C12Q2563/107C12Q2545/113
Inventor 宋祥军祁钊王景芳杨侃侃涂健邵颖刘华朱良强陈曦王勇王振宇
Owner ANHUI AGRICULTURAL UNIVERSITY
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