Method for identifying sex of schistosoma japonicum cercariae with multiplex PCR method

A technology for schistosome cercariae and schistosomiasis, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve problems such as inability to accurately identify cercariae sex, inability to confirm single female cercariae, inability to determine cercariae, etc. Achieve accurate schistosomiasis condition, shorten identification time, and accurately control the effect

Inactive Publication Date: 2019-03-29
SUZHOU UNIV
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Problems solved by technology

However, this method only designs primers for the female-specific amplification region, and cannot accurately identify the sex of cercariae.
For example, for samples without amplified bands, it is impossible to determine that the cercariae escaped from the oncomelania are male cercariae,

Method used

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  • Method for identifying sex of schistosoma japonicum cercariae with multiplex PCR method
  • Method for identifying sex of schistosoma japonicum cercariae with multiplex PCR method

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Embodiment 1

[0040] (1) Sampling of positive oncomelania escaping cercariae: individual positive oncomelania snails were placed in test tubes added with dechlorinated water, and cercariae escaped at room temperature of 26°C under light and other conditions. After 1 hour, pick the cercariae escaped from a single snail and add them to a 1.5ml EP tube. Pick 3 samples from each cercariae escaped from each snail. The DNA of the corresponding cercariae was amplified by multiplex PCR.

[0041](2) DNA extraction of cercariae: put the picked cercariae into a 1.5mL centrifuge tube, add 100μL cercariae lysate (50mM Tris base, 5% Tween20, 50μg / ml proteinase K, pH 7.6-8.0), incubate at 55℃ for 10min Afterwards, bathe in water at 95°C for 1 min, centrifuge at 12,000 r / min for 1 min, and take the supernatant as the DNA template of cercariae.

[0042] (3) Design of multiple PCR primers: a pair of primers were designed for the Z chromosome of Schistosoma japonicum, and a pair of primers were designed for ...

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Abstract

The invention discloses a method for identifying sex of schistosoma japonicum cercariae with a multiplex PCR method and belongs to the technical field of biological detection. A pair of primers is designed for bisexual Z chromosome of schistosoma japonicum, a pair of primers is designed from a characteristic amplification area of a female sequence in W chromosome, and a multiplex PCR system is constructed and can identify male and female cercariae simultaneously. The whole process of identifying the sex of cercariae takes about 4 h with the multiplex PCR method, and the cercaria sex identification time is substantially shortened as compared with existing reports. On one hand, time for establishing a schistosoma japonicum infected animal model is saved, and on the other hand, a required experiment animal model can be established more accurately according to the sex of the cercariae.

Description

technical field [0001] The invention relates to a method for identifying the sex of schistosoma japonicum cercariae by using multiple PCR method, belonging to the technical field of biological detection. Background technique [0002] Schistosoma japonicum is the only trematode with bisexual chromosomes, with 8 pairs of chromosomes, of which the female chromosome is ZW and the male chromosome is ZZ. Although there is obvious dimorphism in the adult stage, females and males can be distinguished by naked eyes, but it is difficult to distinguish males and females in the infection stage, that is, the cercariae stage. Accurate and efficient identification of the sex of schistosomiasis cercariae will help to make a correct evaluation of the genetics, epidemiology and pathological research results of schistosomiasis, and it will be helpful for the research and development of anti-schistosomiasis drugs, accurate evaluation of actual infection and control of schistosomiasis. period i...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6879C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6879C12Q1/6888C12Q2600/16C12Q2537/143
Inventor 许静段中良李春香王小莉刘磊夏超明
Owner SUZHOU UNIV
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