Rapid Discriminative Identification of the Two Predominant Echinococcus Species from Canine Fecal Samples in the Tibetan Region of China by Loop-Mediated Isothermal Amplification–Lateral Flow Dipstick Assay

Xinyue Lv; Jiajia Ai; Xiaojin Mo; Haojie Ding; Sofia Litchev; Entung Lu; Youhong Weng; Qing He; Quzhen Gongsang; Shijie Yang; Xiumin Ma; Jingzhong Li; Huasheng Pang; Shaohong Lu; Qingming Kong

doi:10.3390/tropicalmed9060136

Tropical Medicine and Infectious Disease (Jun 2024)

Rapid Discriminative Identification of the Two Predominant Echinococcus Species from Canine Fecal Samples in the Tibetan Region of China by Loop-Mediated Isothermal Amplification–Lateral Flow Dipstick Assay

Xinyue Lv,
Jiajia Ai,
Xiaojin Mo,
Haojie Ding,
Sofia Litchev,
Entung Lu,
Youhong Weng,
Qing He,
Quzhen Gongsang,
Shijie Yang,
Xiumin Ma,
Jingzhong Li,
Huasheng Pang,
Shaohong Lu,
Qingming Kong

Affiliations

Xinyue Lv: Key Laboratory of Bio-Tech Vaccine of Zhejiang Province, Engineering Research Center of Novel Vaccine of Zhejiang Province, School of Basic Medicine and Forensics, Hangzhou Medical College, Hangzhou 310013, China
Jiajia Ai: Tibet Center for Disease Control and Prevention, NHC Key Laboratory of Echinococcosis Prevention and Control, Lhasa 850000, China
Xiaojin Mo: Tibet Center for Disease Control and Prevention, NHC Key Laboratory of Echinococcosis Prevention and Control, Lhasa 850000, China
Haojie Ding: Key Laboratory of Bio-Tech Vaccine of Zhejiang Province, Engineering Research Center of Novel Vaccine of Zhejiang Province, School of Basic Medicine and Forensics, Hangzhou Medical College, Hangzhou 310013, China
Sofia Litchev: Department of Chemistry & Biochemistry, University of California, Los Angeles, CA 90095, USA
Entung Lu: Santa Monica College, Los Angeles, CA 90405, USA
Youhong Weng: Key Laboratory of Bio-Tech Vaccine of Zhejiang Province, Engineering Research Center of Novel Vaccine of Zhejiang Province, School of Basic Medicine and Forensics, Hangzhou Medical College, Hangzhou 310013, China
Qing He: Key Laboratory of Bio-Tech Vaccine of Zhejiang Province, Engineering Research Center of Novel Vaccine of Zhejiang Province, School of Basic Medicine and Forensics, Hangzhou Medical College, Hangzhou 310013, China
Quzhen Gongsang: Tibet Center for Disease Control and Prevention, NHC Key Laboratory of Echinococcosis Prevention and Control, Lhasa 850000, China
Shijie Yang: National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Chinese Center for Tropical Diseases Research, National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Key Laboratory on Parasite and Vector Biology, Ministry of Health, Shanghai 200025, China
Xiumin Ma: State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, Clinical Laboratory Center, Tumor Hospital Affiliated to Xinjiang Medical University, Urumqi 830000, China
Jingzhong Li: Tibet Center for Disease Control and Prevention, NHC Key Laboratory of Echinococcosis Prevention and Control, Lhasa 850000, China
Huasheng Pang: Tibet Center for Disease Control and Prevention, NHC Key Laboratory of Echinococcosis Prevention and Control, Lhasa 850000, China
Shaohong Lu: Key Laboratory of Bio-Tech Vaccine of Zhejiang Province, Engineering Research Center of Novel Vaccine of Zhejiang Province, School of Basic Medicine and Forensics, Hangzhou Medical College, Hangzhou 310013, China
Qingming Kong: Key Laboratory of Bio-Tech Vaccine of Zhejiang Province, Engineering Research Center of Novel Vaccine of Zhejiang Province, School of Basic Medicine and Forensics, Hangzhou Medical College, Hangzhou 310013, China

DOI: https://doi.org/10.3390/tropicalmed9060136
Journal volume & issue: Vol. 9, no. 6
p. 136

Abstract

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Echinococcosis poses a significant concern in the fields of public health and veterinary care as it can be transmitted between animals and humans. The primary endemic subtypes are cystic echinococcosis (CE) and alveolar echinococcosis (AE), which result from infestation by Echinococcus granulosus and Echinococcus multilocularis, respectively. A prominent epidemic of echinococcosis greatly affects the Tibet Autonomous Region (TAR) in China. A new technique called the loop-mediated isothermal amplification–lateral flow dipstick (LAMP-LFD) test is introduced in this research to differentiate between E. granulosus and E. multilocularis using their repetitive genetic sequences. The test is characterized by its portable nature, simple operation, quick result production, high sensitivity, and low susceptibility to aerosol contamination. The LAMP-LFD method demonstrated an exceptional minimal detection limit, reaching levels as low as approximately 1 fg/μL (femtogram per microliter) of genomic DNA. The assay’s specificity was assessed, and no cross-reactivity was seen. A total of 982 dog fecal samples were collected from 54 counties in the TAR region between July 2021 and June 2022. The established method underwent validation using a commercially available ELISA kit. The agreement rate between the LAMP-LFD and ELISA methods was 97.25%, with a sensitivity of 96.05% and a specificity of 97.35%. The assay described in this study improves specificity by using a double-labeled probe, and it reduces the risk of false-positive results caused by aerosol contamination through the use of a sealed device. This makes it a suitable choice for quickly and accurately identifying the two main types of Echinococcus in field settings.

Published in Tropical Medicine and Infectious Disease

ISSN: 2414-6366 (Online)
Publisher: MDPI AG
Country of publisher: Switzerland
LCC subjects: Medicine
Website: http://www.mdpi.com/journal/tropicalmed

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