Quick detection method for drug susceptibility of coccidium of different species and kit

A technology of drug sensitivity and detection method, applied in the field of bioengineering, can solve the problems of reduced working time and human and material resources, large workload, cumbersome steps, etc., and achieve the effect of saving working time and human and material resources.

Pending Publication Date: 2019-10-11
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention aims to solve the technical problems of cumbersome steps and heavy workload in the current clinical method for detecting the sensitivity of different species of coccidia to vari

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Quick detection method for drug susceptibility of coccidium of different species and kit
  • Quick detection method for drug susceptibility of coccidium of different species and kit
  • Quick detection method for drug susceptibility of coccidium of different species and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1 constructs the qualitative and quantitative analysis method for different species of coccidia to be checked

[0056] 1. The extraction method of genomic DNA is as follows:

[0057] 1. Centrifuge the oocysts to extract the genome (10000rpm, 3min) to remove the potassium dichromate solution, add an appropriate amount of PBS solution (PH=7.4) to the oocyst precipitate, and centrifuge and wash 3 times (3600rpm, 5min);

[0058] 2. Add an equal volume of 425-600 μm glass beads and 250 μL oocyst lysate to the oocyst precipitation, place on a high-throughput tissue disruptor to vibrate and break the wall (60 Hz, 30 s), and wait for more than 90% of the oocysts to rupture. Then add 250 μL oocyst lysate to the centrifuge tube;

[0059] 3. Then add proteinase K (20mg / mL) to the centrifuge tube to a final mass concentration of 100μg / mL, mix well, place in a 50°C water bath and incubate for 3h, and rotate and mix from time to time;

[0060] 4. Add 15 μL of RNaseA (10 ...

Embodiment 2

[0081] Example 2 Clinical Test Sample Collection and Separation

[0082] In chicken farms, use disposable gloves to collect 3-5 portions of fresh feces (>100g), put them in a clean plastic sample box, mark them, and bring them back to the laboratory for inspection. Conventional methods are used to inspect and separate coccidia oocysts. The operation is as follows:

[0083] 1. Take the collected positive stool sample and mix it thoroughly with water at a ratio of 1:9;

[0084] 2. First filter with an 80-mesh sieve, and then use a 200-mesh sieve for secondary filtration to collect the filtrate;

[0085] 3. Let the filtrate stand for 3 hours, carefully discard the upper layer liquid, and leave the precipitate;

[0086]4. Centrifuge the obtained precipitate (1000g, 5min), and discard the supernatant;

[0087] 5. Fully resuspend and mix the precipitate with saturated saline at a volume ratio of 1:5, then centrifuge again (1000g, 5min), keep the supernatant this time, and repeat ...

Embodiment 3

[0091] Clinical effectiveness and sensitivity detection of embodiment 3 diclazuril

[0092] The clinical effectiveness and sensitivity clinical trials of diclazuril anticoccidial drugs refer to the requirements of the guidelines for clinical trials of anticoccidial drugs of the Ministry of Agriculture. The test chickens are about 14 days old healthy chickens, and they are weighed one by one on an empty stomach. Diclazuril was randomly divided into groups with 10 rats in each group. Diclazuril group was used for infection, no drug group for infection and no drug group for non-infection. Each chicken in each infection test group was orally inoculated with 1.5×10 5 sporulated oocysts of a clinically isolated sample to be tested, and according to the instructions for use of diclazuril, 1ppm of diclazuril was added to the feed for continuous feeding and administration, and the non-medication group was fed with no drug-added feed, calculated Evaluation index of clinical effectivene...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a quick detection method for the drug susceptibility of coccidium of different species. The method comprises the following steps of constructing a real-time fluorescence quantification PCR analysis method for TaqMan of the to-be-detected coccidium of different species; collecting clinical test coccidium samples, and separating out coccidium oocysts; carrying out an anti-coccidium drug clinical test; obtaining results of drug susceptibility of the to-be-detected coccidium of different species by analyzing the composition and proportion change of sample coccidium populations before and after the drug test. By means of the quick detection method, the situation in a traditional method that it needs several months before the complicated step of separating a single coccidium is completed can be avoided, the drug susceptibility and the drug resistance test result of the coccidium of different species can be directly obtained in only several days, the working time, labor and material resources are greatly saved, and better prevention and control over clinical coccidiosis are facilitated.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a rapid detection method and a kit for the sensitivity of different species of coccidia to different anticoccidial drugs. Background technique [0002] Coccidiosis is a parasitic disease caused by one or more coccidia parasitizing in different parts such as the intestinal tract of animals, and is characterized by growth retardation and severe damage to the intestinal mucosa. Among them, chicken coccidiosis is a disease caused by one or several single-celled parasitic protozoa parasitic in chicken intestinal epithelial cells caused by Eimeria in the apicomplexan parasite. The most common disease in chicken farms is also the main cause of 6-10% broiler chicken death. Wherever chickens are raised, there is chicken coccidia, and the economic loss caused by chicken coccidia in the world exceeds 3 billion US dollars every year. The economic loss caused by the disease is more th...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/6893C12Q1/686C12Q1/04C12N15/11C12R1/90
CPCC12Q1/6893C12Q1/686C12Q2600/106C12Q2561/113C12Q2563/107C12Q2545/114C12Q2561/101
Inventor 费陈忠张丽芳薛飞群谷峰张可煜王霄旸王米王春梅刘迎春张敏李雪雁
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap