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Loop-mediated isothermal amplification (LAMP) detection method and kit of soft-shelled turtle iridovirus (STIV)

A loop-mediated isothermal, detection kit technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve problems such as non-proliferation

Inactive Publication Date: 2011-08-10
SHENZHEN AUDAQUE DATA TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The virus can proliferate on cell lines such as CO, CK, BF-2, EPC, and FHM, but not in cell lines such as CHSE, R1, RTG-2, and PG

Method used

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  • Loop-mediated isothermal amplification (LAMP) detection method and kit of soft-shelled turtle iridovirus (STIV)
  • Loop-mediated isothermal amplification (LAMP) detection method and kit of soft-shelled turtle iridovirus (STIV)
  • Loop-mediated isothermal amplification (LAMP) detection method and kit of soft-shelled turtle iridovirus (STIV)

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] 1. Materials and Methods

[0050] 1.1 virus

[0051] Turtle iridescent virus comes from: soft-shelled turtle iridescent virus (STIV9701 strain) isolated from soft-shelled turtles in our laboratory. Infectious hematopoietic necrosis virus (IHNV) was kindly provided by Professor B. Hill from the Center for Environmental, Fisheries and Breeding Sciences, UK, and viral hemorrhagic sepsis virus (VHSV) was provided by Dr. Roar Gudding, National Veterinary Research Institute, Norway. Pike Rhabdovirus (PFRV), Infectious Pancreatic Necrosis Virus (IPNV), Fish Viral Neuronecrosis Virus (VNNV), Lymphocyst Virus (LCDV), Singapore Grouper Virus (SGIV), Tiger Frog Virus (TFV) ), Burrough iridescent virus (BIV), etc. are kept by the Aquatic Animal Laboratory of the Animal and Plant Inspection and Quarantine Technology Center of Shenzhen Entry-Exit Inspection and Quarantine Bureau.

[0052] 1.2 LAMP primers

[0053] According to the highly conserved region of the thymidine kinase ge...

Embodiment 2

[0088] A detection kit for soft-shelled turtle iridescent virus, the kit includes the following two pairs of primers:

[0089] Forward Internal Primer (FIP) (Seq ID No.3):

[0090] 5'-TGGAAAAGTCTTCGGGCACCACGCAAGTCCACACTCCTCAG-3';

[0091] Reverse Internal Primer (BIP) (Seq ID No.4):

[0092] 5'-CTGTTTGAGATGGCCCTGGAGGTCTCGCTCTGGATGAGCATA-3'.

[0093] Forward outer primer (F3) (Seq ID No.1):

[0094] 5'-CGTTTAGCGGCAACATTGG-3';

[0095] Reverse outer primer (B3) (Seq ID No.2):

[0096] 5'-TTTCTTTGCGGCCCTCTG-3'.

[0097] The kit also includes instructions for use. This kit can be used for the detection of soft-shelled turtle iridescent virus LAMP.

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Abstract

The invention discloses a loop-mediated isothermal amplification (LAMP) detection method and a kit of soft-shelled turtle iridovirus (STIV). An outer primer comprises sequences shown as Seq ID No.1 and Seq ID No.2. A quick, sensitive and specific detection method for detecting the STIV is provided, field instant detection of viruses can be performed, and technical support is provided for healthy cultivation of soft-shelled turtles and the development of international trade.

Description

technical field [0001] The invention relates to a virus detection method and a detection kit, in particular to a soft-shell turtle iridescent virus (STIV) loop-mediated isothermal amplification (LAMP) detection method and a kit. Background technique [0002] Iridovirus (Iridovirus) is a kind of pathogenic pathogen that is widely infective to fish, amphibians and reptile aquatic animals. The diseases caused by iridovirus have caused huge economic losses to the world's aquaculture industry. In recent years, many countries have successively reported the isolation of iridescent viruses from aquatic economic animals such as diseased fish, frogs and turtles. The family Iridovirudae is divided into 5 genera: Iridovirus, Chloriridovirus, Ranavirus, Lymphocystivirus, Megalocytivirus (Also known as cytomegalovirus). [0003] Soft-shelled turtle iridescent virus (STIV), also known as Chinese soft-shelled turtle virus, belongs to the genus Ranavirus in the family Iridoviridae. The vi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor 何俊强刘宗晓郑晓聪刘荭高隆英兰文升
Owner SHENZHEN AUDAQUE DATA TECH
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