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PCR-RFLP method for distinguishing metacercaria of clonorchis sinensis from metacercaria of metorchis orientalis

A PCR-RFLP, Clonorchis sinensis technology, applied in the PCR-RFLP field of distinguishing Clonorchis sinensis and oriental suborchis cysts, achieves the effects of high stability, high efficiency and accuracy, and a simple method

Inactive Publication Date: 2015-02-25
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0004] At present, there is no report on the use of PCR-RFLP method to distinguish the metacercariae of Clonorchis sinensis and Mesorchis orientalis

Method used

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  • PCR-RFLP method for distinguishing metacercaria of clonorchis sinensis from metacercaria of metorchis orientalis
  • PCR-RFLP method for distinguishing metacercaria of clonorchis sinensis from metacercaria of metorchis orientalis

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Embodiment 1

[0036] figure 1 Two kinds of trematode metacercariae were subjected to ITS gene PCR reaction to obtain about 1200bp electrophoresis;

[0037] Among them: left 1, 2, 3 are Mesorchis orientalis metacercariae (wheat ears) in Zhaodong; left 4, 5, 6 are Mesorchis orientalis metacercariae (willow root); right 1, 2, 3 are Zhaodong Clonorchis sinensis metacercariae (wheat ears), right 4, 5, 6 are Clonorchis sinensis metacercariae (willow root).

[0038] figure 2 Two trematode metacercariae xho I endonuclease digestion analysis result chart;

[0039] Among them: left 1, 2, 3 are Mesorchis orientalis metacercariae (wheat ears) in Zhaodong; left 4, 5, 6 are Mesorchis orientalis metacercariae (willow root); right 1, 2, 3 are Zhaodong Clonorchis sinensis metacercariae (wheat ears), right 4, 5, 6 are Clonorchis sinensis metacercariae (willow root).

[0040] Sample source: Clonorchis sinensis metacercariae and Mesorchis orientalis metacercariae were collected from freshwater fish ...

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Abstract

The invention discloses a PCR-RFLP method for distinguishing metacercaria of clonorchis sinensis from metacercaria of metorchis orientalis, and relates to a molecular method for identifying metacercaria of clonorchis sinensis and metacercaria of metorchis orientalis by using a PCR-RFLP technology. A ribosome transcribed spacer sequence (ITS) of which the length is about 1200bp can be amplified from the metacercaria of clonorchis sinensis and the metacercaria of metorchis orientalis by using the same primers through PCR amplification; the PCR product is subjected to enzyme digestion by virtue of restriction endonuclease XhoI; the sample with two stripes displayed in an electrophoretogram is metacercaria of clonorchis sinensis; and the sample with one stripe displayed in the electrophoretogram is metacercaria of metorchis orientalis. Therefore, accurate and fast identification can be achieved according to the stripe change after enzyme digestion of the PCR product. The identification method disclosed by the invention is simple, relatively high in efficiency and accuracy rate, and fast in identification, only needs a working day, and simultaneously is relatively low in cost.

Description

technical field [0001] The invention relates to a molecular method for distinguishing Clonorchis sinensis metacercariae and Mesorchis orientalis metacercariae by using PCR-RFLP technology, in particular to a PCR method for distinguishing Clonorchis sinensis metacercariae and Mesorchis orientalis metacercariae -RFLP method. Background technique [0002] Clonorchiasis is a very important zoonotic parasitic disease, which is distributed in river basins. Clonorchis sinensis needs two intermediate hosts during the development process. The first intermediate host is freshwater snails, and the second intermediate host is freshwater fish and shrimp. There are about 140 species of freshwater fish hosts confirmed in China, mainly carps. . Clonorchis sinensis adults parasitize the liver and bile ducts of dogs, cats and humans, causing clonorchiasis. Clonorchiasis mainly causes cholangitis, cholecystitis, cholangiohepatitis, and liver cirrhosis, and can even be complicated by hum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/683
Inventor 仇建华陈媛媛高俊峰张莹于剑邵志辉李超丛艳昭齐楷陈瑛琦王春仁
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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