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Universal rpa nucleic acid isothermal amplification primers, kits and applications for bivalve mollusk-infected herpes virus oshv-1

A herpes virus, bivalve mollusk technology, applied in DNA/RNA fragments, microorganisms, recombinant DNA technology, etc., can solve the problem of inability to achieve rapid on-site detection, and achieve good specificity test results, high sensitivity, and good stability. sexual effect

Active Publication Date: 2021-11-09
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the detection process of bivalve molluscs infected with herpes virus OsHV-1 is dependent on instruments and equipment, and rapid on-site detection cannot be achieved

Method used

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  • Universal rpa nucleic acid isothermal amplification primers, kits and applications for bivalve mollusk-infected herpes virus oshv-1
  • Universal rpa nucleic acid isothermal amplification primers, kits and applications for bivalve mollusk-infected herpes virus oshv-1
  • Universal rpa nucleic acid isothermal amplification primers, kits and applications for bivalve mollusk-infected herpes virus oshv-1

Examples

Experimental program
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Embodiment 1

[0037] Example 1: Establishment of the test strip RPA detection method for rapid on-site detection of OsHV-1

[0038] 1. Design of RPA upstream and downstream primers and probe primers

[0039] The genome sequences of OsHV-1 variant strains KP412538.1, GQ153938.1, NC_005881.2, MG561751.2, KY242785.1, KY271630.1 and MF509813.1 were downloaded from NCBI's GenBank and compared and analyzed to determine the differences For the conserved region among OsHV-1 mutant strains, a set of primers was designed for the zero mutation conserved region, in order to detect OsHV-1 of different mutant strains. The present invention designed and synthesized six sets of primers respectively, as shown in Table 1 below.

[0040] Table 1

[0041]

[0042] 2. DNA extraction from clinical samples

[0043]All clinical OsHV-1 positive samples used in the present invention are collected and preserved by our laboratory. The TIANamp Marine Animals DNA Kit from TIANGEN was used to extract the total DNA...

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Abstract

The invention belongs to the technical field of marine biology, and in particular relates to a universal RPA nucleic acid isothermal amplification primer, a kit and an application thereof for bivalve mollusks infected with herpes virus OsHV‑1. The primers are a set of amplification primers, consisting of upstream and downstream primers and probe primers; the upstream primers are shown in SEQ ID NO.1; the downstream primers are shown in SEQ ID NO.2; the probe primers are shown in SEQ ID NO.3 shown. The present invention designs general-purpose RPA amplification detection primers for 7 kinds of OsHV-1 mutant strains in currently known four kinds of bivalve hosts, and establishes a test strip RPA detection system and method for OsHV-1. On-site rapid screening and detection of OsHV‑1 is of great value and has broad market application prospects.

Description

technical field [0001] The invention belongs to the technical field of marine biology, and in particular relates to a universal RPA nucleic acid isothermal amplification primer, a kit and an application thereof for bivalve molluscs infected with herpes virus OsHV-1. Background technique [0002] Ostreid Herpesvirus 1, OsHV-1, which is prevalent in bivalve molluscs, belongs to Herpesvirales (Herpesvirales), Mollusc Herpesviridae (Malacoherpesviridae), OsHV-1 is widely prevalent in coastal shellfish farming or fishing areas around the world . Currently known hosts of bivalve molluscs that can be infected with OsHV-1 mainly include oysters, scallops, and cockles. Frequently led to large-scale death of mud cockles, hair cockles and cockles in coastal areas of my country. There is a certain degree of variation in the infection of OsHV-1 in different species, and the similarity between genomes is about 90%, forming multiple OsHV-1 mutant strains. Establishing a general on-site d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11C12R1/93
CPCC12Q1/6844C12Q1/705C12Q2521/507C12Q2563/131C12Q2565/625
Inventor 辛鲁生于江南白昌明王崇明
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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