Special primers and identification method for rapidly identifying anoplocephalidae tapeworms parasitizing on Equine

A technology of phyllocephalus and phyllocephalus, applied in biochemical equipment and methods, botanical equipment and methods, microbiological measurement/inspection, etc., can solve the problems of inability to infect insect species, diagnosis, inability to distinguish, etc.

Inactive Publication Date: 2014-07-30
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
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  • Claims
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Problems solved by technology

This method cannot distinguish which tapeworm ...

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  • Special primers and identification method for rapidly identifying anoplocephalidae tapeworms parasitizing on Equine
  • Special primers and identification method for rapidly identifying anoplocephalidae tapeworms parasitizing on Equine

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Embodiment 1

[0023] The steps of the method for quickly distinguishing the phyllocephalus parasitic in the genus Equus of the present invention are as follows:

[0024] 1. Sample collection: Collection of gestational proglottids (abbreviated as gestational segments): Taenia parasitic in the digestive tract are often excreted in whole gestational segmental segments. Larger segmental segments in feces are easy to find. For smaller ones, first collect the feces in the basin, add 5-10 times of water, stir evenly, let it stand for natural precipitation, then pour out the upper liquid, add fresh water, stir evenly, and repeat the operation until the upper liquid until clear. Finally, the supernatant was poured out, and the worms were looked for with the naked eye.

[0025] 2. Extraction of the DNA genome: Take about 20 mg of the proglottids prepared in step (1), transfer them into a mortar, and grind vigorously to form a homogenate. Add 350 μL of Buffer PBS (purchased from AXYGEN) and 0.9 μL o...

Embodiment 2

[0034] The difference between this embodiment and embodiment 1 is:

[0035] 1. Sample collection: egg collection: Take 10g of feces, add more than 100mL of water, stir into feces, filter through a 250μm copper sieve, collect the filtrate in a Erlenmeyer flask or beaker, let it settle for 20-40min, and pour off the upper layer. Keep the sediment, add water to mix, and then precipitate, and repeat the operation until the upper liquid is transparent, then pour off the upper liquid. Add 100mL of saturated brine, mix well, and let it stand for half an hour, then the eggs will float up. Pipette the liquid containing worms in the upper layer into another 50mL centrifuge tube, add PBS (pH7.2) to a total volume of 45mL, and centrifuge at 5000×g for 10min. The collected sediment is the eggs. In the prior art, sedimentation or floating methods are used to collect eggs, and the collected eggs can be used for microscope observation. In order to carry out differential diagnosis on the co...

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Abstract

The present invention provides special primers for rapidly identifying anoplocephalidae tapeworms parasitizing on Equine, wherein the upstream primer is 5'-GCAGAGTTTTGCTGATTTTATGAAG-3', and the downstream primer is 5'-CCACTATACTCAGTATTAAATCCACTAACAAG-3'. The present invention further provides a method for rapidly identifying anoplocephalidae tapeworms parasitizing on Equine, wherein the method comprises: adopting the primers to carry out PCR amplification on NAD1 of anoplocephalidae tapeworm, sequencing, comparing the sequencing result with sequences in SEQIDNo.1 and SEQIDNo.2, and judging. With the method, the defects that the general morphology observation can not distinguish gravid proglottids and the microscopic observation can not distinguish eggs are solved. The PCR method of the present invention can be used for molecule identification of two anoplocephalidae tapeworms, and can further be used for epidemiological investigation of the Equine anoplocephalidae tapeworm.

Description

technical field [0001] The present invention relates to a kind of special primer and its identification method for rapid identification of phyllocephalus parasitic on animals of the genus Equus. Background technique [0002] Two species of phyllocephala in the genus Equus: Anoplocephala perfoliata and A. magna parasitize in the large and small intestines of the genus Equus. They are distributed worldwide and have been reported all over my country. Especially in the pastoral areas of Northwest and Inner Mongolia, it is often endemic. The site where the worm parasitizes can cause inflammation and edema of the mucous membrane, damage the mucous membrane, and form a circular hemorrhagic ulcer with tissue hyperplasia. Once the ulcer is perforated, it will cause acute peritonitis and lead to death. When a large number of Pectocephala phyllodes are infected, ulcers are spread all over the gyrus, blind, and colon. Ileocecal stricture obstruction, acute catarrhal enteritis and mucos...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12N15/12
CPCC12Q1/6869C12Q2531/113
Inventor 郭爱疆
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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