Development of LAMP Assays for the Molecular Detection of Taeniid Infection in Canine in Tibetan Rural Area

Overview

Journal J Vet Med Sci

Specialty Veterinary Medicine

Date 2017 Oct 24

PMID 29057765

Citations 6

Authors

Kai Feng

Wei Li

Zhihong Guo

Hong Duo

Yong Fu

Xiuying Shen

Cheng Tie

Rijie E

Changqin Xiao

Yanhong Luo

Guo Qi

Ma Ni

Qingmei Ma

Wataru Yamazaki

Ayako Yoshida

Yoichiro Horii

Kinpei Yagi

Nariaki Nonaka

Affiliations

Soon will be listed here.

Abstract

For field-identification of taeniid cestodes in canine animals in Tibetan area, loop-mediated isothermal amplification (LAMP) assays for Echinococcus multilocularis, E. shiquicus, Taenia hydatigena, T. multiceps, T. pisiformis and T. crassiceps were developed and evaluated along with the reported assay for E. granulosus. The LAMP assays showed specific reaction with their corresponding target species DNA with the detection limit of 1 to 10 pg. Moreover, the assays for E. granulosus, E. multilocularis, T. hydatigena and T. multiceps could detect DNA extracted from 3 or more eggs of their corresponding target species. Then, the LAMP assays were applied on samples containing 3 to 35 taeniid eggs obtained from 61 field-collected canine feces in Qinghai, and the result was compared with a reported multiplex PCR and sequence analysis. The LAMP assays and the PCR detected single species DNA of E. granulosus, E. shiquicus, T. hydatigena and T. multiceps in 5, 2, 44 and 2 samples, respectively. In the rest 8 samples, DNA of both E. granulosus and T. hydatigena were detected by the PCR but the LAMP assays detected those DNAs in 2 samples and only T. hydatigena DNA in 6 samples. It was assumed that less than 3 E. granulosus eggs were mixed in the samples although the samples contained 21 to 27 eggs in total. In conclusion, the LAMP assays were less sensitive than the multiplex PCR, but would have adequate sensitivity for field use in Tibetan area.

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