Primers, kit and identifying method for simultaneously and rapidly identifying 4 types of taenia solium larvae

The technology of a tapeworm and a kit is applied in the field of determination and inspection of microorganisms to achieve the effect of shortening the time of differential diagnosis

Active Publication Date: 2020-01-21
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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Problems solved by technology

[0003] In order to solve the problems existing in the morphological identification of the existing tapeworm larvae, the present invention provides a multiplex method for the differential diagnosis of 4 kinds of tapeworm larvae (cysticercus porcine, cysticercus asiatica, cysticercus lentinococcus and echinococcus granulosus) PCR primers, reagents and identification methods, PCR amplification in the same reaction system, PCR produc...

Method used

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  • Primers, kit and identifying method for simultaneously and rapidly identifying 4 types of taenia solium larvae
  • Primers, kit and identifying method for simultaneously and rapidly identifying 4 types of taenia solium larvae
  • Primers, kit and identifying method for simultaneously and rapidly identifying 4 types of taenia solium larvae

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Embodiment 1

[0034] 1. Alignment analysis of mitochondrial genome sequences of 4 Taenia larvae and design of species-specific primers

[0035] According to the comparative analysis results of the mitochondrial genome sequence blast of 4 species of cestodes (larvae), the following two principles are followed: (1) The primers must have species specificity; The difference is obvious. Design the following 4 specific primers as upstream primers and 1 primer as general downstream primers:

[0036] Cysticercus spp. (ThF): 5′-TAGTTCTTTTGTTAGATGGGTGATTT-3′

[0037] Echinococcus granulosus (EgF): 5′-GCGATTGAACTCATATTGAGC-3′

[0038] Cysticercus asiatica (TaF): 5′-AGGGTTTAAGTTATAAATGTGA-3′

[0039] Cysticercus porcine (TsF): 5′-CTAGGCCACTTAGTAGTTTAGTT-3′

[0040] Universal downstream primer (TahsegR): 5'-CACATAAAACAAAGCCTCAAAC-3'.

[0041] figure 1 Partial sequences of mitochondria and primer positions of 4 species of Taenia larvae.

[0042] 2. Sample Collection

[0043] After the pig is slaugh...

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Abstract

The present invention provides primers for simultaneously and rapidly identifying 4 types of taenia solium larvae of cysticercus cellulosae, taenia asiatica cysticercus, cysticercus tenuicollis and echinococcus granulosus, and also provides a corresponding detection kit and a detection method. The designed specific primers for the taenia solium larvae can quickly identify 4 types of the widely prevalent taenia solium larvae in China in one PCR reaction system, overcome a problem that insect bodies are similar in forms and cannot be accurately identified by morphology, and also overcome a defect of piece weight PCR identification of a single species, besides, PCR products do not need sequencing and sequence alignment, molecular identification of the 4 types of the taenia solium larvae can be realized only according to sizes of electrophoretic fragments of the PCR products, which greatly shortens time of identification diagnosis, and the primers can be used for rapid identification diagnosis of the 4 types of taenia solium larvae and epidemiological investigation of metacestodiasis.

Description

technical field [0001] The invention belongs to the technical field of microbial determination and inspection methods, and in particular relates to primers, kits and identification methods for rapidly identifying four kinds of Taenia larvae at the same time, capable of rapidly identifying cysticerci and Taenia saginata cysts in the same PCR reaction system cercariae, cysticercosis and echinococcus granulosus. Background technique [0002] Taenia solium, Taenia asiatica, Taenia hydatigena and Echinococcus granulosus are all tapeworms of the family Taeniidae. The middle stage larvae (cysticerci or echinococcosis) of Taeniaceae are commonly known as tapeworm larvae. Cysticercus porcine (cysticercosis), cysticercus asiatica, echinococcosis granulosus and cysticercus cerevisiae are all tapeworm larvae parasitic in pigs, and they can all parasitize in the liver of pigs. Cysticercus and Taeniasis cysticercosis) and waste of internal organs (Echinococcosis granulosus), and Cystice...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/16C12Q2537/143C12Q2565/125
Inventor 骆学农郭爱民张少华李艳萍梁盼红王立群才学鹏
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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