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Loop-mediated isothermal amplification (LAMP) method based on cercaria-stage gene of schistosoma japonicum katsurada

A loop-mediated isothermal, schistosome technology, applied in the field of temperature amplification, can solve the problems of low detection specificity and sensitivity, inaccurate detection of schistosomiasis, etc.

Inactive Publication Date: 2017-05-10
HUBEI UNIVERSITY OF MEDICINE
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Problems solved by technology

[0007] The purpose of the present invention is to provide a loop-mediated isothermal amplification method based on the cercarial stage gene of Schistosoma japonicum, which is used to solve the existing problems of inaccurate detection of Schistosoma japonicum and low detection specificity and sensitivity

Method used

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  • Loop-mediated isothermal amplification (LAMP) method based on cercaria-stage gene of schistosoma japonicum katsurada
  • Loop-mediated isothermal amplification (LAMP) method based on cercaria-stage gene of schistosoma japonicum katsurada
  • Loop-mediated isothermal amplification (LAMP) method based on cercaria-stage gene of schistosoma japonicum katsurada

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Embodiment

[0037] Step 1, establishment of Schistosoma japonicum animal model and collection of Schistosoma samples:

[0038] A. Infected snails were incubated in ultra-pure water. The cercariae escaped from the water surface were observed with the naked eye. The cercariae were connected to the inoculation ring and placed on a glass slide. The activity of the cercariae was observed under a dissecting microscope. It was confirmed that the cercariae were active. 10 Kunming mice were taken, and the abdomen was trimmed until With bare skin, take the cover glass and place it on the glass slide, inoculate 30±5 cercariae on the cover glass, wipe the skin of the mouse with cotton balls, stick the glass slide containing the cercariae on the abdominal skin of the mouse for about 1 min, Establish a mouse model infected with schistosomiasis;

[0039] B. After 45 days of infection, the mice were killed by neck dissection, and the pathological changes of the liver and spleen were dissected to observe ...

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Abstract

The invention belongs to the technical field of detection of cercaria-stage gene of schistosoma japonicum katsurada and particularly relates to a loop-mediated isothermal amplification (LAMP) method based on the cercaria-stage gene of the schistosoma japonicum katsurada. The LAMP method comprises the following steps: firstly, establishing a schistosoma japonicum katsurada animal model and collecting a schistosoma japonicum katsurada sample; secondly, preparing a template DNA (Deoxyribonucleic Acid): collecting ultrapure water subjected to oncomelania inoculation, transferring in a centrifugal pipe, and centrifuging under the condition of 1000 to 5000r / min for 5 to 20 minutes; removing supernate, extracting cercaria genome DNA from precipitation by using a blood / cell / tissue genome DNA extraction kit, pulverizing infected oncomelania, collecting oncomelania meat and internal organs, extracting oncomelania genome DNA by using a mollusk genome DNA extraction kit, and putting the extracted cercaria genome DNA and the oncomelania genome DNA at the temperature of 20 DEG C for later use; thirdly, determining a candidate gene: selecting a plurality of predicted protein genes of the cercaria stage of schistosoma japonicum katsurada as target genes; fourthly, designing and synthesizing an LAMP primer; fifthly, configuring the LAMP primer; sixthly, carrying out LAMP detection on the cercaria stage and the infected oncomelania genome DNA.

Description

technical field [0001] The invention belongs to the technical field of detection of genes encoding proteins in the cercarial stage of Schistosoma japonicum, and in particular relates to a loop-mediated isothermal amplification method based on the cercarial stage gene of Schistosoma japonicum. Background technique [0002] Schistosoma japonicum is an important zoonotic disease and one of the most serious public health problems in the world. It is mainly prevalent in 76 countries and regions in the tropics and subtropics. The number of schistosomiasis infections in the world exceeds 200 million people and 600 million people. threatened by infection. There are 453 schistosomiasis endemic counties (cities, districts) with a total population of 252 million; there are 29,980 endemic villages with a total population of 68.613 million. Of the 453 endemic counties (cities, districts) in the country, 343 (accounting for 75.72%) met the standards for interruption of schistosomiasis tr...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6806C12Q1/6844C12Q2531/119
Inventor 李健张轶静杨树国赵燕清关玮
Owner HUBEI UNIVERSITY OF MEDICINE
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