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Schistosoma japonicum katsurada mitochondrial molecular marker and application thereof

A technology of schistosomiasis, mitochondrial gene, applied in the field of biotechnology and genetics

Active Publication Date: 2016-10-05
STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still no effective method for distinguishing regional strains of Schistosoma japonicum, especially the Taiwanese type (ie, non-pathogenic strains) from other regional strains

Method used

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  • Schistosoma japonicum katsurada mitochondrial molecular marker and application thereof
  • Schistosoma japonicum katsurada mitochondrial molecular marker and application thereof
  • Schistosoma japonicum katsurada mitochondrial molecular marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 Sample Collection, Genomic DNA Extraction and Amplification of Target Fragments

[0073] (1) Collect samples of Schistosoma japonicum from 12 endemic areas including Guichi in Anhui, Tongling in Anhui, Yueyang in Hunan, Changde in Hunan, Shashi in Hubei, Duchang in Jiangxi, Xichang in Sichuan, Eryuan in Yunnan, Taiwan, Japan, the Philippines, and Indonesia, Genomic DNA of a single individual was extracted (extraction reagent: DNeasy Blood & Tissue Kit produced by QIAGEN (Germany)). The specific method is as follows:

[0074] Put a single adult Schistosoma japonicum into a 1.5ml EP tube, add 500μl of 1.0mM EDTA and 10mM Tris buffer. After oscillating, suck up the liquid and repeat three times. Add 180 μl of ATL buffer provided in the kit; add 20 μl proteinase K, shake and mix. Incubate in a water bath at 56 °C for 4 h, shaking several times in between. Shake for 15 seconds after taking it out of the water bath, add 200 μl buffer AL, and shake to mix. Then ...

Embodiment 2

[0085] Example 2 Sequence Alignment

[0086]Using MEGA 5.1 software, the gene sequence obtained by sequencing in Example 1 was used for multiple sequence alignment with the reference sequence (database) of known geographical origin in the standard database.

[0087] One of the sequence alignment results (ND1 gene) can be seen figure 1 , it can be seen that there are quite a lot of base variations among individuals in different endemic areas. The other two genes also had base variations among different individuals (results not shown).

Embodiment 3

[0088] Construction of embodiment 3 phylogenetic tree

[0089] Through model analysis software (jmodeltest) evaluation and manual sampling inspection, the most suitable nucleotide evolution model was selected as the HKY model, and the evolutionary tree was constructed based on the selected model by the maximum likelihood method (Maximum Likelihood, ML). According to the topological structure of the phylogenetic tree (especially each branch point and Bootstrap value), the source region of the sample was judged.

[0090] 3.1 The phylogenetic tree construction of a single gene

[0091] 3.1.1 Phylogenetic tree constructed by ND1

[0092] ML phylogenetic tree constructed with ND1 gene, see the results image 3 , it can be seen that the ND1 gene can distinguish Taiwan (TW), Indonesia (IN) and Yunnan (YNEY) very well;

[0093] 3.1.2 Phylogenetic tree constructed by ND4

[0094] ML phylogenetic tree constructed with ND4 gene, see the results Figure 4 , it can be seen that the ND...

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Abstract

The invention provides a schistosoma japonicum katsurada mitochondrial molecular marker and an application thereof. Specifically, the invention discloses a method for identifying geographical isolates of schistosoma japonicum katsurada samples and / or for clustering regional populations of the schistosoma japonicum katsurada samples on the basis of a cluster analysis result of ND1, ND1, ND4 and 16S-12S rDNA gene sequences. The method disclosed by the invention is convenient and accurate and is high in stability; and the method is of great help to identify an original infection place of patients with schistosomiasis and to judge whether a case belongs to a primary type or an imported type. Meanwhile, the method also provides support for monitoring the schistosoma japonicum katsurada and for conducting researches on an epidemic status.

Description

technical field [0001] The invention belongs to the field of biotechnology and genetics, in particular, the invention relates to a method for identifying schistosomiasis populations using mitochondrial genes as molecular markers and the application of mitochondrial molecular markers in population genetics of schistosoma japonicum. Background technique [0002] Schistosomiasis japonicum is a zoonotic disease that seriously threatens human health. About 280,000 people worldwide die from schistosomiasis every year. After decades of hard work, the prevention and control of schistosomiasis in my country has achieved world-renowned achievements. However, there are still about 300,000 active infected people in my country. The situation of schistosomiasis prevention and control is not optimistic, and the task of prevention and control is still arduous. [0003] Schistosoma japonicum has been endemic in Southeast Asia for many years, and has obvious genetic diversity in different end...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 胡薇徐斌冯正殷明波李鸿雁郑鸿翔莫筱瑾
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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