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

J 2021

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

SEARS, William J.; Yvonne QVARNSTROM; Eric DAHLSTROM; Kirsten SNOOK; Lisa KALUNA et al.

Základní údaje

Originální název

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

Autoři

SEARS, William J.; Yvonne QVARNSTROM; Eric DAHLSTROM; Kirsten SNOOK; Lisa KALUNA; Vojtech BALÁŽ; Barbora FECKOVA; Jan ŠLAPETA; David MODRÝ; Susan JARVI a Thomas B. NUTMAN

Vydání

Clinical Infectious Diseases, Oxford University Press, 2021, 1058-4838

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10606 Microbiology

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 20.999

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/21:00123533

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

PCR; Angiostrongylus; eosinophilia; meningitis

Štítky

14314020, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 16. 5. 2022 12:19, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

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.

Citovat

SEARS, William J.; Yvonne QVARNSTROM; Eric DAHLSTROM; Kirsten SNOOK; Lisa KALUNA; Vojtech BALÁŽ; Barbora FECKOVA; Jan ŠLAPETA; David MODRÝ; Susan JARVI a Thomas B. NUTMAN. AcanR3990 qPCR: A Novel, Highly Sensitive, Bioinformatically-Informed Assay to Detect Angiostrongylus cantonensis Infections. Clinical Infectious Diseases. Oxford University Press, 2021, roč. 73, č. 7, s. "E1594"-"E1600", 7 s. ISSN 1058-4838. Dostupné z: https://doi.org/10.1093/cid/ciaa1791.

@article{1817164,
   author = {Sears, William J. and Qvarnstrom, Yvonne and Dahlstrom, Eric and Snook, Kirsten and Kaluna, Lisa and Baláž, Vojtech and Feckova, Barbora and Šlapeta, Jan and Modrý, David and Jarvi, Susan and Nutman, Thomas B.},
   article_number = {7},
   doi = {https://doi.org/10.1093/cid/ciaa1791},
   keywords = {PCR; Angiostrongylus; eosinophilia; meningitis},
   language = {eng},
   issn = {1058-4838},
   journal = {Clinical Infectious Diseases},
   title = {AcanR3990 qPCR: A Novel, Highly Sensitive, Bioinformatically-Informed Assay to Detect Angiostrongylus cantonensis Infections},
   url = {https://doi.org/10.1093/cid/ciaa1791},
   volume = {73},
   year = {2021}
}

TY  - JOUR
ID  - 1817164
AU  - Sears, William J. - Qvarnstrom, Yvonne - Dahlstrom, Eric - Snook, Kirsten - Kaluna, Lisa - Baláž, Vojtech - Feckova, Barbora - Šlapeta, Jan - Modrý, David - Jarvi, Susan - Nutman, Thomas B.
PY  - 2021
TI  - AcanR3990 qPCR: A Novel, Highly Sensitive, Bioinformatically-Informed Assay to Detect Angiostrongylus cantonensis Infections
JF  - Clinical Infectious Diseases
VL  - 73
IS  - 7
SP  - "E1594"-"E1600"
EP  - "E1594"-"E1600"
PB  - Oxford University Press
SN  - 10584838
KW  - PCR
KW  - Angiostrongylus
KW  - eosinophilia
KW  - meningitis
UR  - https://doi.org/10.1093/cid/ciaa1791
N2  - 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.
ER  -

SEARS, William J.; Yvonne QVARNSTROM; Eric DAHLSTROM; Kirsten SNOOK; Lisa KALUNA; Vojtech BALÁŽ; Barbora FECKOVA; Jan ŠLAPETA; David MODRÝ; Susan JARVI a Thomas B. NUTMAN. AcanR3990 qPCR: A Novel, Highly Sensitive, Bioinformatically-Informed Assay to Detect Angiostrongylus cantonensis Infections. \textit{Clinical Infectious Diseases}. Oxford University Press, 2021, roč.~73, č.~7, s.~''E1594''-''E1600'', 7 s. ISSN~1058-4838. Dostupné z: https://doi.org/10.1093/cid/ciaa1791.

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