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Duck tembusu virus nucleic acid CRISPR-Cas13a detection system, RPA primer pair and crRNA

A technology of duck Tembusu virus and Tembusu virus, which is applied in the field of Duck Tembusu virus nucleic acid CRISPR-Cas13a detection system, can solve the problems of complex operation, difficulty in adapting to on-site rapid and accurate detection, and time-consuming, and achieve High sensitivity and good response effect

Pending Publication Date: 2022-04-08
INST OF ANIMAL HUSBANDRY & VETERINARY MEDICINE ANHUI ACAD OF AGRI SCI
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  • Claims
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Problems solved by technology

[0003] At present, the detection of DTMUV usually adopts pathological microscope observation, immunological test, cell culture and PCR detection. Difficult to adapt to the needs of on-site rapid and accurate detection

Method used

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  • Duck tembusu virus nucleic acid CRISPR-Cas13a detection system, RPA primer pair and crRNA
  • Duck tembusu virus nucleic acid CRISPR-Cas13a detection system, RPA primer pair and crRNA
  • Duck tembusu virus nucleic acid CRISPR-Cas13a detection system, RPA primer pair and crRNA

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Embodiment 1

[0067] A kind of RPA primer pair that adopts CRISPR-Cas13a detection system to detect duck Tembusu virus nucleic acid that the present invention proposes, wherein, RPA primer pair is selected from any one pair in following 5 pairs of primer pairs:

[0068] (1) A primer pair consisting of RPA F1 and RPA R1, wherein the nucleotide sequence of RPA F1 and the nucleotide sequence of RPA R1 are shown in Table 1;

[0069] (2) A primer pair consisting of RPA F2 and RPA R2, wherein the nucleotide sequence of RPA F2 and the nucleotide sequence of RPA R2 are shown in Table 1;

[0070] (3) A pair of primers consisting of RPA F3 and RPA R3, wherein the nucleotide sequence of RPA F3 and the nucleotide sequence of RPA R3 are shown in Table 1;

[0071] (4) A pair of primers consisting of RPA F4 and RPA R4, wherein the nucleotide sequence of RPA F4 and the nucleotide sequence of RPA R4 are shown in Table 1;

[0072] (5) A primer pair consisting of RPA F5 and RPA R5, wherein the nucleotide seq...

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Abstract

The invention relates to the technical field of duck tembusu virus nucleic acid detection, particularly discloses a duck tembusu virus nucleic acid CRISPR-Cas13a detection system, an RPA primer pair and crRNA, and establishes a visual, sensitive and specific duck tembusu virus rapid clinical detection system based on a CRISPR-Cas13a diagnosis platform based on the designed RPA primer pair and the specific crRNA. The system does not need a complex temperature control instrument in the whole reaction process, isothermal detection only needs to be carried out at 37 DEG C, the system can be used for fluorescence reading or visual reading, the lowest detection limit of the detection system is 1.0 * 100 copies / reaction, cross reaction with other poultry viruses is avoided, the Cas13a detection system can also directly play a role in virus detection of clinical samples, and the Cas13a detection system can be used for detecting the viruses of the poultry. The kit has the advantages of simplicity, convenience and rapidness in detection operation, suitability for field detection, high specificity, high detection sensitivity, high accuracy and reliability.

Description

technical field [0001] The invention relates to the technical field of duck Tembusu virus nucleic acid, more specifically, it relates to a duck Tembusu virus nucleic acid CRISPR-Cas13a detection system, RPA primer pair and crRNA. Background technique [0002] Duck tembusu virus (DTMUV) belongs to the family Fiaviviridae, the genus Flavivirus, and the Ntaya virus group, and is related to dengue virus (DENV), West Nile virus (WNV) ), Japanese encephalitis virus (JEV) and Zika virus (ZAKV) are members of the same virus genus. Duck Tembusu virus can cause a large drop in egg production of breeding ducks and laying ducks, from the egg production peak or high egg production rate to 20% to 30% rapidly, and in serious cases, production will be stopped about 7 days after the onset. Therefore, rapid and timely detection of pathogens is an effective means to reduce their spread. [0003] At present, the detection of DTMUV usually adopts pathological microscope observation, immunologi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11C12N15/113
CPCY02A50/30
Inventor 戴银殷冬冬尹磊王洁茹胡晓苗程帮照王骏俊沈学怀潘孝成赵瑞宏
Owner INST OF ANIMAL HUSBANDRY & VETERINARY MEDICINE ANHUI ACAD OF AGRI SCI
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