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Primer set for testing mud crab bicistronic mRNA virus, test method and rapid diagnosis kit

A bicistronic and rapid diagnosis technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/testing, etc., can solve problems such as the detection of Scylla bicistronic virus that has not yet been seen, Achieve the effect of reducing background influence, high detection sensitivity and high specificity

Inactive Publication Date: 2013-12-25
SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to its own advantages, LAMP has been widely used in the detection and diagnosis of pathogenic microorganisms, including viruses, bacteria, fungi, parasites and other infectious diseases. Related reports on anti-virus detection

Method used

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  • Primer set for testing mud crab bicistronic mRNA virus, test method and rapid diagnosis kit
  • Primer set for testing mud crab bicistronic mRNA virus, test method and rapid diagnosis kit
  • Primer set for testing mud crab bicistronic mRNA virus, test method and rapid diagnosis kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Scylla Bicistronic Virus Detection Primer Set and the Detection Method of Scylla Bicistronic Virus

[0070] 1.1 Primer set for detection of Scylla bicistronic virus

[0071] In this example, according to the published Scylla bicistronic virus genome sequence, the specific sequence of the Scylla bicistronic virus gene was selected, and then analyzed and designed to be used in the LAMP detection technology, which can specifically identify the Scylla bicistronic virus gene sequence. The specific primer set of cistron virus, this primer set is made up of following four primers:

[0072] Outer primer F3, the nucleotide sequence of which is shown in SEQ ID NO:1;

[0073] Outer primer B3, the nucleotide sequence of which is shown in SEQ ID NO:2;

[0074] Inner primer FIP, its nucleotide sequence is shown in SEQ ID NO:3;

[0075] Internal primer BIP, its nucleotide sequence is shown in SEQ ID NO:4.

[0076] 1.2 Detection method of Scylla bicistronic virus

[0077...

Embodiment 2

[0079] Example 2 Scylla Bicistronic Virus Rapid Diagnostic Kit

[0080] The rapid diagnostic kit of this embodiment can be used for the rapid diagnosis of whether the sample contains Scylla bicistronic virus by the loop-mediated isothermal amplification technique. , Bst DNA polymerase, positive control substance and chromogenic solution, etc., wherein:

[0081] (1) Primer set for Scylla bicistronic virus detection: DNA synthesizer, 4 tubes, containing the nucleotide sequence of outer primer F3, outer primer B3, inner primer FIP and inner primer BIP, and outer primer F3 respectively As shown in SEQ ID NO:3; The nucleotide sequence of outer primer B3 is shown in SEQ ID NO:4; The nucleotide sequence of inner primer FIP is shown in SEQ ID NO:1; The nucleotide sequence of inner primer BIP The sequence is shown in SEQ ID NO: 2. The storage concentration of the above four primers is 10 μM respectively; the final concentration of the reaction is: 0.8 μM FIP, 0.8 μM BIP, 0.2 μM F3, 0....

Embodiment 3

[0096] Example 3 Scylla Bicistronic Virus Rapid Diagnostic Kit

[0097] The rapid diagnostic kit of this embodiment can be used for the rapid diagnosis of whether the sample contains Scylla bicistronic virus by the loop-mediated isothermal amplification technique. , Bst DNA polymerase, positive control substance, chromogenic solution, Trizol, chloroform, isopropanol, 75% ethanol by volume, RNase-free water, RT reaction solution, reverse transcriptase and foam plate. The kit provides reagents for the reverse transcription step. in:

[0098] (1) Primer set for Scylla bicistronic virus detection: DNA synthesizer, 4 tubes, containing the nucleotide sequence of outer primer F3, outer primer B3, inner primer FIP and inner primer BIP, and outer primer F3 respectively As shown in SEQ ID NO:3; The nucleotide sequence of outer primer B3 is shown in SEQ ID NO:4; The nucleotide sequence of inner primer FIP is shown in SEQ ID NO:1; The nucleotide sequence of inner primer BIP The sequenc...

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Abstract

The invention discloses a primer set for testing mud crab bicistronic mRNA virus, a test method and a rapid diagnosis kit. The primer set includes the following four primers: outer primer F3, outer primer B3, inner primer FIP and inner primer BIP. The primer set for testing mud crab bicistronic mRNA virus, the test method and the rapid diagnosis kit can test mud crab bicistronic mRNA virus at high efficiency and high specificity, the specificity of the primer set is high, and the test method has the advantages of low test cost, short time consumption, high productivity, high specificity, remarkable color difference between negative and positive results and high verification rate, and is clearer and more reliable; and the kit in the invention establishes a set of optimized inverse transcription-loop-mediated isothermal amplification reaction system, can be used for tracking and testing mud crab dbicistronic mRNA virus at each stage in the process of culturing blue crabs, and can prevent the spread of virus, so the efficiency of scientific management is increased, and the high practical value is realized.

Description

technical field [0001] The invention relates to a primer set for detecting scylla bicistronic virus, a detection method and a rapid diagnosis kit. Background technique [0002] Scylla paramamosain (formerly identified as Scylla serrata) is an important aquaculture species in my country's marine fishery because of its delicious meat, rapid growth and wide distribution. However, with the gradual expansion of the breeding scale, the disease problem is also becoming more and more serious. Mud Crab Dicistrovirus (MCDV) is a newly discovered pathogen in the infected mud crab in recent years. The blue crabs infected with MCDV have no obvious change in body color, loss of appetite, lack of activity, and will not dive into the sand even during the day, which can lead to high mortality. The virus belongs to the single positive-sense RNA, Dicistroviridae family. Due to the late discovery, there are few related studies. MCDV can infect blue crabs through immersion, fasting immersion...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 郭志勋张迪冯娟苏友禄徐力文
Owner SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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