Angie-LAMP for diagnosis of human eosinophilic meningitis using dog as proxy: A LAMP assay for Angiostrongylus cantonensis DNA in cerebrospinal fluid
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 host...
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| Veröffentlicht in: | PLoS neglected tropical diseases 2023-05, Vol.17 (5), p.e0011038-e0011038 |
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| creator | Baláž, Vojtech Rivory, Phoebe Hayward, Douglas Jaensch, Susan Malik, Richard Lee, Rogan Modrý, David Šlapeta, Jan |
| description | 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.
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.
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. |
| doi_str_mv | 10.1371/journal.pntd.0011038 |
| format | Article |
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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.
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.</description><identifier>ISSN: 1935-2735</identifier><identifier>ISSN: 1935-2727</identifier><identifier>EISSN: 1935-2735</identifier><identifier>DOI: 10.1371/journal.pntd.0011038</identifier><identifier>PMID: 37126515</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Accessibility ; Angiostrongylus ; Angiostrongylus - genetics ; Angiostrongylus cantonensis ; Angiostrongylus cantonensis - genetics ; Animals ; Assaying ; Biology and Life Sciences ; Cerebrospinal fluid ; Deoxyribonucleic acid ; Diagnosis ; DNA ; DNA, Ribosomal ; Dogs ; Engineering and Technology ; Genetic aspects ; Haplotypes ; Health aspects ; Humans ; Indigenous species ; Jurisdiction ; Laboratories ; Laboratory tests ; Larvae ; Leukocytes (eosinophilic) ; Lysis ; Medicine and Health Sciences ; Meningitis ; Meningitis - diagnosis ; Meningitis - veterinary ; Nucleotide sequence ; PCR ; Physical sciences ; Pulmonary arteries ; Rats ; Research and analysis methods ; Samples ; Sequencing ; Snails - genetics ; Strongylida Infections - diagnosis ; Strongylida Infections - veterinary ; Tropical diseases</subject><ispartof>PLoS neglected tropical diseases, 2023-05, Vol.17 (5), p.e0011038-e0011038</ispartof><rights>Copyright: © 2023 Baláž et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</rights><rights>COPYRIGHT 2023 Public Library of Science</rights><rights>2023 Baláž et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2023 Baláž et al 2023 Baláž et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c559t-73af5c11bd750cc43e20d4943c6ed32a2dcfe5347f9adace3fe3bd7952ba6c533</citedby><cites>FETCH-LOGICAL-c559t-73af5c11bd750cc43e20d4943c6ed32a2dcfe5347f9adace3fe3bd7952ba6c533</cites><orcidid>0000-0003-1459-9117</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC10174499/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC10174499/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2928,23866,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/37126515$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Nzelu, Chukwunonso</contributor><creatorcontrib>Baláž, Vojtech</creatorcontrib><creatorcontrib>Rivory, Phoebe</creatorcontrib><creatorcontrib>Hayward, Douglas</creatorcontrib><creatorcontrib>Jaensch, Susan</creatorcontrib><creatorcontrib>Malik, Richard</creatorcontrib><creatorcontrib>Lee, Rogan</creatorcontrib><creatorcontrib>Modrý, David</creatorcontrib><creatorcontrib>Šlapeta, Jan</creatorcontrib><title>Angie-LAMP for diagnosis of human eosinophilic meningitis using dog as proxy: A LAMP assay for Angiostrongylus cantonensis DNA in cerebrospinal fluid</title><title>PLoS neglected tropical diseases</title><addtitle>PLoS Negl Trop Dis</addtitle><description>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.
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.
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.</description><subject>Accessibility</subject><subject>Angiostrongylus</subject><subject>Angiostrongylus - genetics</subject><subject>Angiostrongylus cantonensis</subject><subject>Angiostrongylus cantonensis - genetics</subject><subject>Animals</subject><subject>Assaying</subject><subject>Biology and Life Sciences</subject><subject>Cerebrospinal fluid</subject><subject>Deoxyribonucleic acid</subject><subject>Diagnosis</subject><subject>DNA</subject><subject>DNA, Ribosomal</subject><subject>Dogs</subject><subject>Engineering and Technology</subject><subject>Genetic aspects</subject><subject>Haplotypes</subject><subject>Health aspects</subject><subject>Humans</subject><subject>Indigenous species</subject><subject>Jurisdiction</subject><subject>Laboratories</subject><subject>Laboratory tests</subject><subject>Larvae</subject><subject>Leukocytes (eosinophilic)</subject><subject>Lysis</subject><subject>Medicine and Health Sciences</subject><subject>Meningitis</subject><subject>Meningitis - diagnosis</subject><subject>Meningitis - veterinary</subject><subject>Nucleotide sequence</subject><subject>PCR</subject><subject>Physical sciences</subject><subject>Pulmonary arteries</subject><subject>Rats</subject><subject>Research and analysis methods</subject><subject>Samples</subject><subject>Sequencing</subject><subject>Snails - genetics</subject><subject>Strongylida Infections - diagnosis</subject><subject>Strongylida Infections - veterinary</subject><subject>Tropical diseases</subject><issn>1935-2735</issn><issn>1935-2727</issn><issn>1935-2735</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><recordid>eNptkluP1CAUxxujcS_6DYySmGx86QilTFtfTLNek_HyoM-EgUOHDYUKrXE-iN9XutvdzJgND9x-__85HE6WPSN4RWhFXl_5KThhV4Mb1QpjQjCtH2SnpKEsLyrKHh6sT7KzGK8wZg2ryePsJOmLNSPsNPvbus5Avmm_fEfaB6SM6JyPJiKv0W7qhUOQts4PO2ONRD04kxRjAqZ03CHlOyQiGoL_s3-DWnTtJGIU-2u_2d7HMXjX7e0UkRRu9A7cHOHd1xYZhyQE2AYfB5Oeg7SdjHqSPdLCRni6zOfZzw_vf1x-yjffPn6-bDe5ZKwZ84oKzSQhW1UxLGVJocCqbEoq16BoIQolNTBaVroRSkigGmhiG1ZsxVoySs-zFze-g_WRLxWNvKiLdY1rgkki3i7EtO1BSXBjEJYPwfQi7LkXhh_fOLPjnf_Nk7gqy6ZJDq8Wh-B_TRBH3psowVrhwE9zMFwXpCzxOqEv_0PvT2mhOmGBG6d9CixnU95WjNAGM1YnanUPlYaC3sj0Bdqk8yPBxYFgB8KOu-jtNBrv4jFY3oAyfVoMoO-qQTCfW_M2az63Jl9aM8meH1byTnTbi_QfH5vi1g</recordid><startdate>202305</startdate><enddate>202305</enddate><creator>Baláž, Vojtech</creator><creator>Rivory, Phoebe</creator><creator>Hayward, Douglas</creator><creator>Jaensch, Susan</creator><creator>Malik, Richard</creator><creator>Lee, Rogan</creator><creator>Modrý, David</creator><creator>Šlapeta, Jan</creator><general>Public Library of Science</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7SS</scope><scope>7T2</scope><scope>7T7</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8C1</scope><scope>8FD</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>F1W</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>H94</scope><scope>H95</scope><scope>H97</scope><scope>K9.</scope><scope>L.G</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>P64</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0003-1459-9117</orcidid></search><sort><creationdate>202305</creationdate><title>Angie-LAMP for diagnosis of human eosinophilic meningitis using dog as proxy: A LAMP assay for Angiostrongylus cantonensis DNA in cerebrospinal fluid</title><author>Baláž, Vojtech ; Rivory, Phoebe ; Hayward, Douglas ; Jaensch, Susan ; Malik, Richard ; Lee, Rogan ; Modrý, David ; Šlapeta, Jan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c559t-73af5c11bd750cc43e20d4943c6ed32a2dcfe5347f9adace3fe3bd7952ba6c533</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Accessibility</topic><topic>Angiostrongylus</topic><topic>Angiostrongylus - genetics</topic><topic>Angiostrongylus cantonensis</topic><topic>Angiostrongylus cantonensis - genetics</topic><topic>Animals</topic><topic>Assaying</topic><topic>Biology and Life Sciences</topic><topic>Cerebrospinal fluid</topic><topic>Deoxyribonucleic acid</topic><topic>Diagnosis</topic><topic>DNA</topic><topic>DNA, Ribosomal</topic><topic>Dogs</topic><topic>Engineering and Technology</topic><topic>Genetic aspects</topic><topic>Haplotypes</topic><topic>Health aspects</topic><topic>Humans</topic><topic>Indigenous species</topic><topic>Jurisdiction</topic><topic>Laboratories</topic><topic>Laboratory tests</topic><topic>Larvae</topic><topic>Leukocytes (eosinophilic)</topic><topic>Lysis</topic><topic>Medicine and Health Sciences</topic><topic>Meningitis</topic><topic>Meningitis - diagnosis</topic><topic>Meningitis - veterinary</topic><topic>Nucleotide sequence</topic><topic>PCR</topic><topic>Physical sciences</topic><topic>Pulmonary arteries</topic><topic>Rats</topic><topic>Research and analysis methods</topic><topic>Samples</topic><topic>Sequencing</topic><topic>Snails - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>PLoS neglected tropical diseases</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Baláž, Vojtech</au><au>Rivory, Phoebe</au><au>Hayward, Douglas</au><au>Jaensch, Susan</au><au>Malik, Richard</au><au>Lee, Rogan</au><au>Modrý, David</au><au>Šlapeta, Jan</au><au>Nzelu, Chukwunonso</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Angie-LAMP for diagnosis of human eosinophilic meningitis using dog as proxy: A LAMP assay for Angiostrongylus cantonensis DNA in cerebrospinal fluid</atitle><jtitle>PLoS neglected tropical diseases</jtitle><addtitle>PLoS Negl Trop Dis</addtitle><date>2023-05</date><risdate>2023</risdate><volume>17</volume><issue>5</issue><spage>e0011038</spage><epage>e0011038</epage><pages>e0011038-e0011038</pages><issn>1935-2735</issn><issn>1935-2727</issn><eissn>1935-2735</eissn><abstract>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.
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.
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.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>37126515</pmid><doi>10.1371/journal.pntd.0011038</doi><orcidid>https://orcid.org/0000-0003-1459-9117</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1935-2735 |
| ispartof | PLoS neglected tropical diseases, 2023-05, Vol.17 (5), p.e0011038-e0011038 |
| issn | 1935-2735 1935-2727 1935-2735 |
| language | eng |
| recordid | cdi_plos_journals_2826808101 |
| source | MEDLINE; Public Library of Science; DOAJ Directory of Open Access Journals; PubMed Central; EZB Electronic Journals Library; PubMed Central Open Access |
| subjects | Accessibility Angiostrongylus Angiostrongylus - genetics Angiostrongylus cantonensis Angiostrongylus cantonensis - genetics Animals Assaying Biology and Life Sciences Cerebrospinal fluid Deoxyribonucleic acid Diagnosis DNA DNA, Ribosomal Dogs Engineering and Technology Genetic aspects Haplotypes Health aspects Humans Indigenous species Jurisdiction Laboratories Laboratory tests Larvae Leukocytes (eosinophilic) Lysis Medicine and Health Sciences Meningitis Meningitis - diagnosis Meningitis - veterinary Nucleotide sequence PCR Physical sciences Pulmonary arteries Rats Research and analysis methods Samples Sequencing Snails - genetics Strongylida Infections - diagnosis Strongylida Infections - veterinary Tropical diseases |
| title | Angie-LAMP for diagnosis of human eosinophilic meningitis using dog as proxy: A LAMP assay for Angiostrongylus cantonensis DNA in cerebrospinal fluid |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-20T19%3A18%3A48IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Angie-LAMP%20for%20diagnosis%20of%20human%20eosinophilic%20meningitis%20using%20dog%20as%20proxy:%20A%20LAMP%20assay%20for%20Angiostrongylus%20cantonensis%20DNA%20in%20cerebrospinal%20fluid&rft.jtitle=PLoS%20neglected%20tropical%20diseases&rft.au=Bal%C3%A1%C5%BE,%20Vojtech&rft.date=2023-05&rft.volume=17&rft.issue=5&rft.spage=e0011038&rft.epage=e0011038&rft.pages=e0011038-e0011038&rft.issn=1935-2735&rft.eissn=1935-2735&rft_id=info:doi/10.1371/journal.pntd.0011038&rft_dat=%3Cgale_plos_%3EA751390558%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2826808101&rft_id=info:pmid/37126515&rft_galeid=A751390558&rfr_iscdi=true |