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Ora serrata crab reovirus diagnostic reagent kit and its inspection

A technology of reovirus and diagnostic kits, applied in biochemical equipment and methods, measurement/inspection of microorganisms, material analysis through observation of the impact on chemical indicators, etc., can solve the problem that there is no effective treatment for viral diseases and other issues, to achieve the effect of avoiding virus transmission, high practical value, and high sensitivity

Inactive Publication Date: 2007-04-04
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, there is no effective treatment for viral diseases

Method used

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  • Ora serrata crab reovirus diagnostic reagent kit and its inspection

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Experimental program
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Embodiment 1

[0030] Embodiment 1: The genetic diagnosis kit of Scylla serrata reovirus

[0031] The kit consists of the following parts (10 samples):

[0032] 1. RNA Extraction Solution A, 2 tubes, 5ml / tube, filled with Trizol solution.

[0033] 2. RNA Extraction Solution B, 1 tube, filled with chloroform, 2ml in total.

[0034] 3. RNA Extraction Solution C, 1 tube, filled with isopropanol, 5ml in total.

[0035] 4. RNA Extraction Solution D, 1 tube, filled with 70% ethanol, 10ml in total.

[0036] 5. RNA extraction solution E solution, 1 tube, 0.5ml / tube, filled with DEPC water.

[0037] 6. RT reaction solution (solution F), 1 tube, 0.1ml / tube, containing RT reaction solution, including 5×Buffer, dNTP, DEPC-H 2 O, random primers, RNase inhibitor, reverse transcriptase M-MLV (RT).

[0038] 7. RT-PCR reaction solution (solution G), 1 tube, 0.2ml / tube, containing RT-PCR reaction solution, including 10×Buffer (including mg 2- ), dNTP, forward primer F, reverse primer R, ddH 2 O and Taq...

Embodiment 2

[0059] Example 2: Detection method of Scylla serrata reogu

[0060] Use the test kit of embodiment 1, carry out according to the following steps:

[0061] 1. Using aseptic operation method, take 0.1g of fresh Scylla serrata gill tissue and add 1ml of liquid A, homogenize in an ice bath in a homogenizer, and let stand at room temperature for 3-5 minutes.

[0062] 2. Add 200 μl of solution B to the above homogenate, mix by inverting up and down, and let stand for 5 minutes.

[0063] 3. Centrifuge at 12000r / min for 10min at 4°C.

[0064] 4. Take 400 μl of supernatant, add an equal volume of solution C, shake gently, and let stand for 10 minutes.

[0065] 5. Centrifuge at 12000r / min for 15min at 4°C.

[0066] 6. Discard the supernatant, wash twice with 1ml of pre-cooled solution D, and centrifuge at 7500r / min for 5min at 4°C.

[0067] 7. Air-dry or blow-dry on an ultra-clean workbench, add 50 μl E liquid water to dissolve (if it cannot be completely dissolved, it can be placed...

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Abstract

A kit used for diagnosing reovirus of juyuanqin crab consists of a pair of primer designed according to RNA sequence of a reovirus separated out from ill juyuanqin crab. Its detection method utilizes reverse transcription - polymerase chain reaction technique to carry out qualitative detection on specific RNA nucleic acid section of reovirus on juyuanqin crab.

Description

technical field [0001] The invention relates to a diagnostic kit, in particular to a diagnostic kit for aquatic economic animal diseases and a detection method thereof. Background technique [0002] Scylla serrata has delicious meat and rich nutrition. It is an important aquatic economic animal in my country. It is widely cultivated in coastal areas of Guangdong, Fujian, Hainan, Zhejiang and other provinces. However, with the increase of breeding scale and density, the disease is becoming more and more serious, causing great economic losses. Pathogenic organisms that cause diseases include bacteria, viruses, parasites, etc., and there are many types. From May to November 2004, an epidemic occurred in Zhuhai City, Guangdong Province, with a mortality rate of over 70%. The study found that reovirus played an important role in it. So far, there is no effective treatment for viral diseases. Preventing virus infection and prevalence is the main measure that can be taken. Earl...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N21/77C12Q1/68C12Q1/70
Inventor 郭志勋翁少萍何建国刘利利
Owner SUN YAT SEN UNIV
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