AcanR3990 qPCR: A Novel, Highly Sensitive, Bioinformatically-Informed Assay to Detect <i>Angiostrongylus cantonensis</i> Infections

Sears, William J.; Qvarnström, Yvonne; Dahlstrom, Eric; Snook, Kirsten; Kaluna, Lisa; Baláž, Vojtěch; Fecková, Barbora; Šlapeta, Jan; Modrý, David; Jarvi, Susan I.; Nutman, Thomas B.

doi:10.1093/cid/ciaa1791

Cited by 29 publications

(52 citation statements)

References 27 publications

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“…Angiostrongylus cantonensis DNA was detected with replication in nearly every tissue tested using the AcanR3990 qPCR assay including heart septum, lung, all nine samples from brain, CSF, and blood tissues, while the AcanITS1 assay [2,11] detected DNA in ve of nine brain samples and CSF. Both PCR assays have been shown to amplify DNA from A. cantonensis as well as from a closely related species A. mackarrasae [15]. While there currently are no PCR-based tests available that distinguish between these two species, detection in this study is likely restricted to A. cantonensis, as A. mackarrasae has yet to be documented in Hawai`i.…”

Section: Discussionmentioning

confidence: 87%

“…Using an AcanITS1 assay developed for qPCR [2], A. cantonensis DNA was detected in the peripheral blood of rats at various time points including very early PI [14]. Recent development of a novel qPCR assay (AcanR3990) based on tandem repeats in genomic DNA has been shown to be ~100-1,000 times more sensitive than the AcanITS1 assay [15]. In Sears et al [15], the AcanR3990 assay detected A. cantonensis DNA in CSF, blood, and brain tissues from a horse exhibiting clinical symptoms of infection, whereas the AcanITS1 assay detected A. cantonensis DNA only in the brain.…”

Section: Introduction Methods and Resultsmentioning

confidence: 99%

“…Recent development of a novel qPCR assay (AcanR3990) based on tandem repeats in genomic DNA has been shown to be ~100-1,000 times more sensitive than the AcanITS1 assay [15]. In Sears et al [15], the AcanR3990 assay detected A. cantonensis DNA in CSF, blood, and brain tissues from a horse exhibiting clinical symptoms of infection, whereas the AcanITS1 assay detected A. cantonensis DNA only in the brain. Here, we apply both the AcanITS1 and the AcanR3990 assays for the detection of A. cantonensis DNA in CSF, various blood products, and multiple organs from a domestic rabbit unintentionally infected through apparent consumption of contaminated local lettuce in a household in Hawai`i.…”

Section: Introduction Methods and Resultsmentioning

confidence: 99%

“…Another aliquot of the same DNA sample was used as template in quantitative PCR using the AcanR3990 assay as described with minor modi cation [15]. Brie y, qPCR was performed in 10 µL reactions containing 1X TaqMan Fast Advanced Master Mix (ThermoFisher Scienti c, Waltham MA), and 5 µL template DNA.…”

Section: Introduction Methods and Resultsmentioning

confidence: 99%

“…The AcanR3990 assay appears more sensitive than the AcanITS1 assay in detecting A. cantonensis DNA from lung, heart septum, brain, and peripheral blood products collected from this rabbit. Angiostrongylus cantonensis DNA in peripheral blood has previously been detected in rats using the AcanITS1 assay [14] and in a horse using the AcanR3990 assay [15]. The AcanR3990 assay has been shown to be up to 1000 times more sensitive than the AcanITS1 assay [15].…”

Section: Discussionmentioning

confidence: 99%

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Autochthonous Transmission of Angiostrongylus Cantonensis in a Domestic Rabbit (Oryctolagus Cuniculus) in Hawai`i: Detection in Multiple Tissues Using AcanITS1 and AcanR3990 PCR Assays

Jarvi

Snook

Mina³

et al. 2022

Preprint

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Background: Hawaii is the hotspot for rat lungworm disease (angiostrongyliasis) caused by the nematode Angiostrongylus cantonensis in the USA. In humans, PCR of the CSF is typically used for diagnosis, however, collection of CSF requires hospitalization. Here, we evaluate the efficacy of two different PCR tests to detect A.cantonensis DNA in multiple tissues including blood from a rabbit presumably infected by eating contaminated lettuce. Methods: Two different PCR assays (AcanR3990, and AcanITS1) were used comparatively to test DNA extracted from slug and rabbit tissues. Assays were conducted using established protocols and were run in triplicate, with negative (dH20) controls included throughout. Results: A juvenile Parmarian martensi (semi-slug) found in local lettuce tested positive for the presence of Angiostrongylus cantonensis DNA. A family and their two domestic rabbits (Oryctolagus cuniculus) consumed this lettuce twice within the five days preceding testing. One rabbit exhibited symptoms consistent with eosinophilic meningitis 3-6 days after being fed the lettuce. Appropriate veterinary treatment was ineffective and the rabbit was subsequently euthanized. This study comparatively applies two different PCR assays to detect A. cantonensis DNA in the peripheral blood, cerebrospinal fluid, brain, heart, and lung tissue of this rabbit, and provides data implicating parasite transmission via contaminated home-grown lettuce. Six of the nine brain DNA samples, as well as the CSF sample, tested positive in replicate or triplicate for A. cantonensis DNA with both PCR assays. The AcanR3990 assay also detected A. cantonensis DNA from the lung, heart septum, all nine samples from the brain, and blood products (plasma, EDTA-treated whole blood, and buffy coat/red blood cells) in replicate or triplicate.

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Section: Discussionmentioning

confidence: 87%

Section: Introduction Methods and Resultsmentioning

confidence: 99%

Section: Introduction Methods and Resultsmentioning

confidence: 99%

Section: Introduction Methods and Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 3 more Smart Citations

Autochthonous Transmission of Angiostrongylus Cantonensis in a Domestic Rabbit (Oryctolagus Cuniculus) in Hawai`i: Detection in Multiple Tissues Using AcanITS1 and AcanR3990 PCR Assays

Jarvi

Snook

Mina³

et al. 2022

Preprint

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Angiostrongylus cantonensis (the Rat Lungworm) Infection and Hearing Loss

Kundi,

Alaskarov,

Ulusoy

2023

Comprehensive ENT

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One-health approach on the future application of snails: a focus on snail-transmitted parasitic diseases

Pathak,

Luitel,

Utaaker

et al. 2023

Parasitol Res

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Snails are fascinating molluscs with unique morphological and physiological adaptive features to cope with various environments. They have traditionally been utilized as food and feed sources in many regions of the world. The future exploitation of alternative nutrient sources, like snails, is likely to increase further. Snails, however, also serve as an intermediate host for several zoonotic parasites. A category of parasitic infections, known as snail-transmitted parasitic diseases (STPDs), is harmful to humans and animals and is mainly driven by various trematodes, cestodes, and nematodes. The environment plays a crucial role in transmitting these parasites, as suitable habitats and conditions can facilitate their growth and proliferation in snails. In light of diverse environmental settings and biologically categorized snail species, this review evaluates the dynamics of significant STPDs of zoological importance. Additionally, possible diagnostic approaches for the prevention of STPDs are highlighted. One-health measures must be considered when employing snails as an alternative food or feed source to ensure the safety of snail-based products and prevent any adverse effects on humans, animals, and the environment.

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AcanR3990 qPCR: A Novel, Highly Sensitive, Bioinformatically-Informed Assay to Detect Angiostrongylus cantonensis Infections

Cited by 29 publications

References 27 publications

Autochthonous Transmission of Angiostrongylus Cantonensis in a Domestic Rabbit (Oryctolagus Cuniculus) in Hawai`i: Detection in Multiple Tissues Using AcanITS1 and AcanR3990 PCR Assays

Autochthonous Transmission of Angiostrongylus Cantonensis in a Domestic Rabbit (Oryctolagus Cuniculus) in Hawai`i: Detection in Multiple Tissues Using AcanITS1 and AcanR3990 PCR Assays

Angiostrongylus cantonensis (the Rat Lungworm) Infection and Hearing Loss

One-health approach on the future application of snails: a focus on snail-transmitted parasitic diseases

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