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Method for detecting medicine resistance of schistosoma japonicum cercariae to praziquantel in vitro

A technology for schistosomiasis cercariae and in vitro detection, which is applied in the direction of biological testing, material inspection products, etc., can solve the problems of drug resistance, long time, missed detection and treatment, etc., and achieve the effect of solving on-site detection and monitoring, and solving technical problems

Inactive Publication Date: 2012-06-27
JIANGSU INST OF PARASITIC DISEASES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Long-term and large-scale repeated application of the same insecticide will often lead to drug resistance, so it is necessary to establish on-site monitoring and detection technology for praziquantel resistance
The current main method of detecting praziquantel resistance is to treat patients on the spot in the epidemic area and evaluate the results of chemotherapy. This method takes a long time, the sequence of patients is low, and there is a phenomenon of missed detection and treatment.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] Example 1: Determination of sensitivity of cercariae of Schistosoma japonicum praziquantel-sensitive strains and resistant strains to praziquantel

[0010] Take a 24-well cell culture plate and add a concentration of 10 -5 mol / L praziquantel solution 1 mL, then use an inoculation loop to transfer 20-50 freshly escaped cercariae of Schistosoma japonicum praziquantel-susceptible and resistant strains, and observe the cercariae in each well 20 min later under a dissecting microscope The change of tail docking was used to calculate the docking rate of cercariae. The experiment was repeated 6 times.

[0011] Table 1 Effects of praziquantel on the changes of cercariae docking in praziquantel-sensitive and resistant strains of Schistosoma japonicum

[0012] Insect strain Action time (min) Concentration of praziquantel solution (mol / L) Docking rate of cercariae (%) Schistosoma japonicum praziquantel-sensitive strain 20 10 -5 68.7±8.9 Praziquantel...

Embodiment 2

[0013] Example 2: Sensitivity determination of cercariae of Schistosoma japonicum praziquantel-sensitive and resistant strains to praziquantel

[0014] Take a 24-well cell culture plate and add a concentration of 5×10 -6 mol / L praziquantel solution 1 mL, then use an inoculation loop to transfer 20-50 freshly escaped cercariae of Schistosoma japonicum praziquantel-susceptible and resistant strains, and observe the cercariae in each well 20 min later under a dissecting microscope The change of tail docking was used to calculate the docking rate of cercariae. The experiment was repeated 6 times.

[0015] Table 2 Effects of praziquantel on the change of cercariae docking in praziquantel-sensitive and resistant strains of Schistosoma japonicum

[0016] Insect strain Action time (min) Concentration of praziquantel solution (mol / L) Docking rate of cercariae (%) Schistosoma japonicum praziquantel-sensitive strain 20 5×10 -6 65.4±9.7 Praziquantel-resista...

Embodiment 3

[0017] Example 3: Sensitivity determination of cercariae of Schistosoma japonicum praziquantel-sensitive and resistant strains to praziquantel

[0018] Take a 24-well cell culture plate and add a concentration of 10 -6 mol / L praziquantel solution 1 mL, then use an inoculation loop to transfer 20-50 freshly escaped cercariae of Schistosoma japonicum praziquantel-susceptible and resistant strains, and observe the cercariae in each well 20 min later under a dissecting microscope The change of tail docking was used to calculate the docking rate of cercariae. The experiment was repeated 6 times.

[0019] Table 3 Effects of praziquantel on the changes of cercariae docking in praziquantel-sensitive and resistant strains of Schistosoma japonicum

[0020] Insect strain Action time (min) Concentration of praziquantel solution (mol / L) Docking rate of cercariae (%) Schistosoma japonicum praziquantel-sensitive strain 20 10 -6 43.8±7.0 Praziquantel-resistant ...

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Abstract

The invention relates to a method for detecting the medicine resistance of schistosoma japonicum cercariae to praziquantel in vitro, and belongs to the technical field of antiparasitic medicines. The method comprises the following steps of: adding 1mL of 10<-5> mol / L praziquantel liquor, 1mL of 5*10<-6> mol / L praziquantel liquor, 1mL of 10<-6> mol / L praziquantel liquor and 1mL of 5*10<-7> mol / L praziquantel liquor into plate holes of a 24-hole cell culture plate, immigrating 20 to 50 newly escaped schistosoma japonicum cercariae by using an inoculating loop, observing the docking change of the cercariae in the holes under an anatomical lens after the praziquantel liquor acts for 20 minutes, and calculating the docking rate of the cercariae, wherein the docking rate of the cercariae is less than 40 percent after 10<-5> mol / L praziquantel acts, is less than 35 percent after 5*10<-6> mol / L praziquantel acts, is less than 30 percent after 10<-6> mol / L praziquantel acts, and is less than 10 percent after 5*10<-7> mol / L praziquantel acts, so that the schistosoma japonicum cercariae have medicine resistance to the praziquantel. The method is quick, simple, convenient, sensitive and reliable and is suitable for quickly detecting and monitoring the medicine resistance of the schistosoma japonicum cercariae on site.

Description

technical field [0001] The invention discloses a method for in vitro detection of resistance of Schistosoma japonicum cercariae to praziquantel, which belongs to the technical field of antiparasitic drugs. Background technique [0002] Schistosomiasis japonicum is a major infectious disease that endangers human health and affects social and economic development. Praziquantel is the only commercially available drug currently used to treat schistosomiasis japonicum. Long-term and large-scale repeated application of the same insecticide will often lead to drug resistance, so it is necessary to establish on-site monitoring and detection technology for praziquantel resistance. The current main method of detecting praziquantel resistance is to treat patients on-site in the epidemic area and evaluate the results of chemotherapy. This method takes a long time, the sequence of patients is low, and there is a phenomenon of missed detection and treatment. We have established an in vi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50
Inventor 李洪军梁幼生汪伟曲国立邢云天戴建荣
Owner JIANGSU INST OF PARASITIC DISEASES
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