Method for preparing positive oncomelania of schistosoma

A technology for schistosomiasis and schistosomiasis, which is applied in the field of bioengineering, can solve problems such as difficult operation, and achieve the effect of efficient in vitro culture and operation

Inactive Publication Date: 2011-09-14
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the oncomelania, the intermediate host of Schistosoma japonicum, is small and difficult to handle, there are few reports on the in vitro manipulation of Schistosoma japonicum larvae

Method used

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  • Method for preparing positive oncomelania of schistosoma
  • Method for preparing positive oncomelania of schistosoma
  • Method for preparing positive oncomelania of schistosoma

Examples

Experimental program
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Effect test

Embodiment 1

[0029] The artificial preparation of embodiment 1 schistosomiasis-positive oncomelania

[0030] 1. Method steps

[0031] 1.1 Schistosoma japonicum egg hatching miracidium and its co-culture with sf9 cells

[0032] Take the liver of a mouse infected with Schistosoma japonicum for 42 days, weigh it and add 4 times the volume of dechlorinated water to homogenate it. Add 20ml of liver homogenate into a 500ml long-necked flat-bottomed flask. In order to avoid contamination, add penicillin and streptomycin to the dechlorinated water for hatching. Chlorine water, incubated at a constant temperature of 20°C-37°C. After 2 hours, take the hatching water from the upper layer of cotton and place it on ice for 2 hours, and centrifuge at 5000 g for 5 minutes to enrich the miracidia.

[0033] Sf9 cells (Spodoptera frugiperda ovarian cells) were cultured and subcultured at 27°C. One day before incubation, the sf9 cells were subcultured into a six-well plate, and 3ml of fresh SF-900 was add...

Embodiment 2

[0055] Example 2 Identification of activity and biological characteristics of cercariae released from schistosomiasis-positive snails

[0056] The cercariae released from the schistosomiasis-positive snails prepared in Example 1 and the cercariae released from the wild snails were patched on the abdomen of 5 Kunming mice respectively, and 40 ± 2 cercariae were quantitatively infected. After 42 days of infection, the portal vein was flushed out by cardiac perfusion. - Mesenteric vein and liver worms, observe the size of the worms, and record the number of worms. Part of the cercariae was prepared with reference to Fang Zhengming’s method of staining slide specimens, and histochemical staining was performed using carmine hydrochloride staining solution (Fang Zhengming. An improved method for making stained slide specimens of Schistosoma cercariae. Chinese Journal of Parasitic Disease Control, 1999, 12 (4 ): 255). The collected cercariae were extracted using the DNAezsol-E micro...

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Abstract

The invention discloses a method for preparing a positive oncomelania of schistosoma. The method comprises the following steps: co-culturing myracidium incubated by schistosoma eggs and sf9 cells to obtain mother sporocyst developed by the myracidium; injecting the mother sporocyst into a negative oncomelania body under microscopy; and continuously feeding the oncomelania containing injected mother sporocyst, and detecting whether the oncomelania is positive or not to obtain the positive oncomelania of the schistosoma. By the method for preparing the positive oncomelania of the schistosoma, cercaria released by the cultured positive oncomelania has the same infectivity and the same biological characteristics with the schistosoma. The method provides an efficient and convenient in vitro manual operation method for studying the genetic functions of the schistosoma.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a preparation method of schistosome-positive oncomelania. Background technique [0002] The growth and development of Schistosoma japonicum is relatively complex, including the alternation of sexual generations in the final host and asexual generations in the intermediate host Oncomelania snails. Schistosoma japonicum has different physiological characteristics in different developmental stages. In view of the harmfulness and widespread nature of Schistosoma japonicum, exploring the development process of Schistosoma japonicum, especially the genes involved in development, is particularly important for understanding the relationship between the host and Schistosoma japonicum, as well as for the development of therapeutic drugs and vaccine research. Many studies use modern science and technology, such as gene silencing, RNA interference, etc., to study the gene function of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K61/00
CPCY02A40/81
Inventor 朱传刚江艳陆珂李小红刘毅石耀军林矫矫
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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