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Method and kit for quickly detecting clonorchiasis sinensis metacercaria in freshwater fish

A technology of Clonorchis sinensis and Clonorchis sinensis, which is applied to the field of rapid detection of Clonorchis sinensis metacercariae in freshwater fish, can solve the problems of complex methods and low sensitivity, and achieve the effect of rapid sensitivity

Inactive Publication Date: 2011-08-10
STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The object of the present invention is to provide a kind of method and kit thereof for rapid detection of C. The technical problem that the detection method of C. sinensis metacercariae in freshwater fish is complicated and the sensitivity is not high

Method used

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  • Method and kit for quickly detecting clonorchiasis sinensis metacercaria in freshwater fish
  • Method and kit for quickly detecting clonorchiasis sinensis metacercaria in freshwater fish
  • Method and kit for quickly detecting clonorchiasis sinensis metacercaria in freshwater fish

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Embodiment 2

[0032] Embodiment 2 traditional kit method

[0033] The main steps of the kit method are:

[0034] 1. Sterilize the ultra-clean table with alcohol, sterilize with ultraviolet light; sterilize the tweezers and scissors; take a sterilized 1.5ml centrifuge tube, add worms (or metacercariae), wash with single distilled water repeatedly, and finally discard the lotion .

[0035] 2. Pre-adjust the thermostat: 55 degrees Celsius; add to the sample tube: 200 microliters of Nuclei analysis solution; 50 microliters of 0.5M EDTA (PH8.0); 20 microliters of Proteinase K (20mg / ml); RNase A Solution (4mg / ml) 5 microliters.

[0036] 3. Heat at 55 degrees Celsius for 16-18 hours, add 250 microliters of lysis Buffer, vortex, and immediately transfer to minicolumns (prepared in advance).

[0037] 4. Centrifuge at 13000 rpm for 3 minutes and 30 seconds, pour off the liquid in the collection tube, and put the column back into the collection tube.

[0038] 5. Add 650 microliters of wizard SV w...

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Abstract

The invention provides a method for quickly detecting clonorchiasis sinensis metacercaria in freshwater fish, which comprises the following steps of: treating an in vitro freshwater fish sample; extracting DNA of the sample; performing polymerase chain reaction (PCR) amplification on the DNA of the sample; and performing electrophoresis observation on the PCR amplification product. In the step of treating the freshwater fish sample, the sample to be detected is added into buffer solution and broken by using glass beads for homogenizing, and then the homogenized solution is sucked and dripped on a filter paper (FTA) card; and in the step of extracting the DNA of the sample, the FTA card is naturally dried, and then the FTA card is washed by using pure terephthalic acid (PTA) card lotion. The invention also provides a kit for quickly detecting the clonorchiasis sinensis metacercaria in the freshwater fish. One clonorchiasis metacercaria in each gram of freshwater fish can be detected by using the method; quantitative PCR technology can distinguish three different metacercaria; and the method is simple, convenient and quick, has high sensitivity, and is suitable for food safety inspection and customs epidemic prevention inspection.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a detection method for Clonorchis sinensis, in particular to a method for rapidly detecting Clonorchis sinensis metacercariae in freshwater fish and a kit thereof. Background technique: [0002] Clonorchiasis sinensis, also known as liver fluke, is a food-borne zoonotic parasitic disease mainly caused by clonorchiasis parasitizing in the liver and bile ducts of humans. Clonorchis sinensis infection has been reported in China, Japan, Korea, Philippines, Singapore, Malaysia, Russia and other countries. In recent years, due to the diversification of food and the introduction of foreign eating habits, the infection rate of this disease in my country has gradually increased, and the infection rate of the population ranges from 1 to 30%. According to the first national survey of parasitic diseases, the national average infection rate is 0.356%. Based on this calculation, there are about ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 陈韶红李树清张永年陈家旭艾琳
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT