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Kit for detecting echinococcus multilocularis pathogen from dog excrement

A technology of echinococcus multilocularis and a kit, which is applied in the field of detection kits for detecting whether there is Echinococcus atrioculum pathogen in dog feces, which can solve the problems of infecting people and the surrounding environment, low sensitivity and specificity, and achieves sensitive Highly specific, highly specific, and easy-to-operate effects

Active Publication Date: 2013-06-26
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] However, the existing detection methods such as conventional etiological detection methods (arecoline catharsis method and autopsy method), stool DNA-PCR detection method and stool antigen ELISA detection method (Dinkel A, et al. Detection of Echinococcus multilocularis in the definitive host: coprodiagnosis by PCR as an alternative to Necropsy[J]. J Clin Microbiol, 1998, 36(7): 1871-1876.; Eckert J. Predictive values ​​and quality control of techniques for the diagnosis of Echinococcus multilocularis in definitive hosts.Acta Trop, 2003, 85(2): 157-163.) have certain shortcomings, the sensitivity and specificity are all low, and the routine detection method also has the risk of infecting people and the surrounding environment

Method used

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  • Kit for detecting echinococcus multilocularis pathogen from dog excrement
  • Kit for detecting echinococcus multilocularis pathogen from dog excrement
  • Kit for detecting echinococcus multilocularis pathogen from dog excrement

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Embodiment Construction

[0013] Below in conjunction with embodiment description.

[0014] In this example, the kit used to detect Echinococcus multilocularis pathogen from dog feces includes: 2×LAMP reaction solution, a mixture composed of four primers, Bst DNA polymerase, standard positive template and chromogenic reagent , where the four specific primers are: upstream external primer F3: 5'-gTgTgTTgCTATATTgCTTgT-3' (SEQ №1); downstream external primer B3: 5'-AACTTTAACAACATACACCTAgT-3' (SEQ №2); upstream internal primer FIP : 5'-TTAACCAACCAATAACAACCCAgTgAATTCgTggTgTTAgTTATTTggTTAgg-3' (SEQ No. 3) and downstream internal primer BIP: 5'-ATgTgACgTTTggTgTggTAgTTAgAATTCAAgAACCACCAAAATAATgTCT-3' (SEQ No. 4), wherein:

[0015] 2×LAMP reaction solution includes: 40mmol / L Tris-HCl (pH 8.8), 20mmol / L KCl, 16mmol / L MgSO 4 , 20mmol / L (NH 4 ) 2 SO 4 , 0.2% (v / v) Tween 20, 1.6mol / L Betaine and 2.5mmol / L dNTP;

[0016] The primer mixture includes: 40pmol / L upstream internal primer FIP, 40pmol / L downstream int...

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Abstract

The invention discloses an LAMP (Loop-Mediated Isotheral Amplification) test kit for detecting the presence of an echinococcus multilocularis pathogen in dog excrement and application thereof. Thet kit for detecting the echinococcus multilocularis pathogen in dog excrement disclosed by the invention comprises four specific primers, i.e., an upstream external primer SEQ No., a downstream external primer SEQ No.2, an upstream internal primer SEQ No.3 and a downstream internal primer SEQ No.4. The test kit can be used for rapidly detecting the presence of the echinococcus multilocularis pathogeny in dog excrement, and has the characteristics of high sensitivity, easiness and convenience for operating, high specificity, rapidness, and the like.

Description

technical field [0001] The invention relates to a detection kit and the use of the kit. Specifically, the invention relates to a detection kit for detecting whether there is Echinococcus multilocularis pathogen in dog feces and the use of the kit. Background technique [0002] Adults of Echinococcus multilocularis mainly parasitize in the small intestines of dogs, wolves, foxes and other canids. The eggs are excreted with feces, infect humans and various rodents, and parasitize their liver and other organs, causing alveolar echinococcosis. Coccidiosis. Human multilocular echinococcosis is a serious hazard with a high fatality rate, also known as "worm cancer", which seriously threatens the life and health of the herdsmen in western my country. Therefore, the establishment of a rapid and effective detection kit for Echinococcus multilocularis-infected canine feces DNA is of great significance for the prevention, control and treatment of the disease. [0003] However, the ex...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 贾万忠倪兴维闫鸿斌娄忠子李宏民
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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