BACKGROUND: Angiostrongylus cantonensis (Ac), or the rat lungworm, is a major cause of eosinophilic meningitis. Humans are infected by ingesting the 3rd stage larvae from primary hosts, snails, and slugs, or paratenic hosts. The currently used molecular test is a qPCR assay targeting the ITS1 rDNA region (ITS1) of Ac. METHODS: In silico design of a more sensitive qPCR assay was performed based on tandem repeats predicted to be the most abundant by the RepeatExplorer algorithm. Genomic DNA (gDNA) of Ac were used to determine the analytical sensitivity and specificity of the best primer/probe combination. This assay was then applied to clinical and environmental samples. RESULTS: The limit of detection of the best performing assay, AcanR3990, was 1 fg (the DNA equivalent of 1/100 000 dilution of a single 3rd stage larvae). Out of 127 CDC archived CSF samples from varied geographic locations, the AcanR3990 qPCR detected the presence of Ac in 49/49 ITS1 confirmed angiostrongyliasis patients, along with 15/73 samples previously negative by ITS1 qPCR despite strong clinical suspicion for angiostrongyliasis. Intermediate hosts (gastropods) and an accidental host, a symptomatic horse, were also tested with similar improvement in detection observed. AcanR3990 qPCR did not cross-react in 5 CSF from patients with proven neurocysticercosis, toxocariasis, gnathostomiasis, and baylisascariasis. AcanR3990 qPCR failed to amplify genomic DNA from the other related Angiostrongylus species tested except for Angiostrongylus mackerrasae (Am), a neurotropic species limited to Australia that would be expected to present with a clinical syndrome indistinguishable from Ac. CONCLUSION: These results suggest AcanR3990 qPCR assay is highly sensitive and specific with potential wide applicability as a One Health detection method for Ac and Am.

• MeSH
• Angiostrongylus cantonensis * genetika   MeSH
• Angiostrongylus * MeSH
• infekce hlísticemi řádu Strongylida * diagnóza   MeSH
• koně MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• meningitida * MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

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.

• MeSH
• Angiostrongylus cantonensis * genetika   MeSH
• Angiostrongylus * genetika   MeSH
• hlemýždi genetika   MeSH
• infekce hlísticemi řádu Strongylida * diagnóza   veterinární   MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• meningitida * diagnóza   veterinární   MeSH
• psi MeSH
• ribozomální DNA MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• centrální nervový systém parazitologie   MeSH
• experimenty na zvířatech MeSH
• helmintóza komplikace   patofyziologie   MeSH
• krysa rodu rattus MeSH
• oči parazitologie   MeSH
• Check Tag
• krysa rodu rattus MeSH

BACKGROUND: Rats (Rattus spp.) invaded most of the world as stowaways including some that carried the rat lungworm, Angiostrongylus cantonensis, the cause of eosinophilic meningoencephalitis in humans and other warm-blooded animals. A high genetic diversity of A. cantonensis based on short mitochondrial DNA regions is reported from Southeast Asia. However, the identity of invasive A. cantonensis is known for only a minority of countries. The affordability of next-generation sequencing for characterisation of A. cantonensis genomes should enable new insights into rat lung worm invasion and parasite identification in experimental studies. METHODS: Genomic DNA from morphologically verified A. cantonensis (two laboratory-maintained strains and two field isolates) was sequenced using low coverage whole genome sequencing. The complete mitochondrial genome was assembled and compared to published A. cantonensis and Angiostrongylus malaysiensis sequences. To determine if the commonly sequenced partial cox1 can unequivocally identify A. cantonensis genetic lineages, the diversity of cox1 was re-evaluated in the context of the publicly available cox1 sequences and the entire mitochondrial genomes. Published experimental studies available in Web of Science were systematically reviewed to reveal published identities of A. cantonensis used in experimental studies. RESULTS: New A. cantonensis mitochondrial genomes from Sydney (Australia), Hawaii (USA), Canary Islands (Spain) and Fatu Hiva (French Polynesia), were assembled from next-generation sequencing data. Comparison of A. cantonensis mitochondrial genomes from outside of Southeast Asia showed low genetic diversity (0.02-1.03%) within a single lineage of A. cantonensis. Both cox1 and cox2 were considered the preferred markers for A. cantonensis haplotype identification. Systematic review revealed that unequivocal A. cantonensis identification of strains used in experimental studies is hindered by absence of their genetic and geographical identity. CONCLUSIONS: Low coverage whole genome sequencing provides data enabling standardised identification of A. cantonensis laboratory strains and field isolates. The phenotype of invasive A. cantonensis, such as the capacity to establish in new territories, has a strong genetic component, as the A. cantonensis found outside of the original endemic area are genetically uniform. It is imperative that the genotype of A. cantonensis strains maintained in laboratories and used in experimental studies is unequivocally characterised.

• MeSH
• Angiostrongylus cantonensis genetika   MeSH
• cyklooxygenasa 1 genetika   MeSH
• fylogeneze MeSH
• genetická variace * MeSH
• genom mitochondriální * MeSH
• genom u helmintů MeSH
• infekce hlísticemi řádu Strongylida parazitologie   MeSH
• krysa rodu rattus MeSH
• mitochondriální DNA * MeSH
• sekvenční analýza DNA MeSH
• sekvenování celého genomu MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Austrálie MeSH
• Havajské ostrovy MeSH
• Polynésie MeSH
• Španělsko MeSH

Lungworms in the genus Angiostrongylus cause disease in animals and humans. The spread of Angiostrongylus vasorum within Europe and the recent establishment of Angiostrongylus cantonensis increase the relevance of these species to veterinary and medical practitioners, and to researchers in parasitology, epidemiology, veterinary science and ecology. This review introduces the key members of the genus present in Europe and their impacts on health, and updates the current epidemiological situation. Expansion of A. vasorum from localized pockets to wide distribution across the continent has been confirmed by a rising prevalence in foxes and increasing reports of infection and disease in dogs, while the list of carnivore and mustelid definitive hosts continues to grow. The tropically distributed rat lungworm A. cantonensis, meanwhile, has been recorded on islands south of Europe, previously the Canary Islands, and now also the Balearic Islands, although so far with limited evidence of zoonotic disease. Other members of the genus, namely, A. chabaudi, A. daskalovi and A. dujardini, are native to Europe and mainly infect wildlife, with unknown consequences for populations, although spill-over can occur into domestic animals and those in zoological collections. The epidemiology of angiostrongylosis is complex, and further research is needed on parasite maintenance in sylvatic hosts, and on the roles of ecology, behaviour and genetics in disease emergence. Improved surveillance in animals and humans is also required to support risk assessments and management.

• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Učebnice pro lékařské a nelékařské studijní programy.

• MeSH
• parazitologie MeSH

• Konspekt
• Mikrobiologie
• NLK Obory
• parazitologie
• NLK Publikační typ
• učebnice vysokých škol

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• alergologie a imunologie
• urgentní lékařství
• infekční lékařství

Tato kniha je určena všem zájemcům o lékařskou mikrobiologii, především ale studentům jak pregraduálním, tak postgraduálním. Kniha je zaměřena spíše lékařsky než mikrobiologicky. Důraz je položen na patogenitu mikrobů a na patogenezi procesů, které v makroorganismu vyvolávají.

• MeSH
• bakteriologie MeSH
• mikrobiologie MeSH
• mykologie MeSH
• parazitologie MeSH
• virologie MeSH
• Publikační typ
• monografie MeSH

• Konspekt
• Mikrobiologie
• NLK Obory
• mikrobiologie, lékařská mikrobiologie
• infekční lékařství