J 2023

Angie-LAMP for diagnosis of human eosinophilic meningitis using dog as proxy: A LAMP assay for Angiostrongylus cantonensis DNA in cerebrospinal fluid

BALÁŽ, Vojtech, Phoebe RIVORY, Douglas HAYWARD, Susan JAENSCH, Richard MALIK et. al.

Basic information

Original name

Angie-LAMP for diagnosis of human eosinophilic meningitis using dog as proxy: A LAMP assay for Angiostrongylus cantonensis DNA in cerebrospinal fluid

Authors

BALÁŽ, Vojtech, Phoebe RIVORY, Douglas HAYWARD, Susan JAENSCH, Richard MALIK, Rogan LEE, David MODRÝ (203 Czech Republic, guarantor, belonging to the institution) and Jan ŠLAPETA

Edition

PLOS Neglected Tropical Diseases, San Francisco, Public Library of Science, 2023, 1935-2735

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

30310 Parasitology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

Impact factor

Impact factor: 3.800 in 2022

RIV identification code

RIV/00216224:14310/23:00132146

Organization unit

Faculty of Science

DOI

UT WoS

000984025300006

Keywords in English

Angiostrongyliasis; Angiostrongylus; Angiostrongylus cantonensis; Animals; DNA; Ribosomal; Dogs; Humans; LAMP assay; Meningitis; Rats; Snails; Strongylida Infections

Tags

Tags

International impact, Reviewed
Změněno: 7/12/2023 08:34, Mgr. Marie Šípková, DiS.

Abstract

V originále

Background Angiostrongylus cantonensis (rat lungworm) is recognised as the leading cause of human eosinophilic meningitis, a serious condition observed when nematode larvae migrate through the CNS. Canine Neural Angiostrongyliasis (CNA) is the analogous disease in dogs. Both humans and dogs are accidental hosts, and a rapid diagnosis is warranted. A highly sensitive PCR based assay is available but often not readily accessible in many jurisdictions. An alternative DNA amplification assay that would further improve accessibility is needed. This study aimed to assess the diagnostic utility of a newly designed LAMP assay to detect DNA of globally distributed and invasive A. cantonensis and Angiostrongylus mackerrasae, the other neurotropic Angiostrongylus species, which is native to Australia. Methodology/Principal findings Cerebrospinal fluid (CSF) from dogs with a presumptive diagnosis of A. cantonensis infection (2020–2022) were received for confirmatory laboratory testing and processed for DNA isolation and ultrasensitive Angiostrongylus qPCR targeting AcanR3390. A newly designed LAMP assay targeting the same gene target was directly compared to the reference ultrasensitive qPCR in a diagnostic laboratory setting to determine the presence of A. cantonensis DNA to diagnose CNA. The LAMP assay (Angie-LAMP) allowed the sensitive detection of A. cantonensis DNA from archived DNA specimens (Kappa = 0.81, 95%CI 0.69–0.92; n = 93) and rapid single-step lysis of archived CSF samples (Kappa = 0.77, 95%CI 0.59–0.94; n = 52). Only A. cantonensis DNA was detected in canine CSF samples, and co-infection with A. mackerrasae using amplicon deep sequencing (ITS-2 rDNA) was not demonstrated. Both SYD.1 and AC13 haplotypes were detected using sequencing of partial cox1. Conclusions/Significance The Angie-LAMP assay is a useful molecular tool for detecting Angiostrongylus DNA in canine CSF and performs comparably to a laboratory Angiostrongylus qPCR. Adaptation of single-step sample lysis improved potential applicability for diagnosis of angiostrongyliasis in a clinical setting for dogs and by extension, to humans.