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Development of a recombinase polymerase amplification (RPA-EXO) and lateral flow assay (RPA-LFA) based on the ITS1 gene for the detection of Angiostrongylus cantonensis in gastropod intermediate hosts

Published online by Cambridge University Press:  04 November 2020

Susan I. Jarvi Open the ORCID record for Susan I. Jarvi [Opens in a new window] ,
Elizabeth S. Atkinson ,
Lisa M. Kaluna ,
Kirsten A. Snook  and
Argon Steel Open the ORCID record for Argon Steel [Opens in a new window]
Susan I. Jarvi*
Affiliation:
Department of Pharmaceutical Sciences, Daniel K. Inouye College of Pharmacy, University of Hawai‘i at Hilo, Hilo, Hawai‘i, USA
Elizabeth S. Atkinson
Affiliation:
Department of Pharmaceutical Sciences, Daniel K. Inouye College of Pharmacy, University of Hawai‘i at Hilo, Hilo, Hawai‘i, USA
Lisa M. Kaluna
Affiliation:
Department of Pharmaceutical Sciences, Daniel K. Inouye College of Pharmacy, University of Hawai‘i at Hilo, Hilo, Hawai‘i, USA
Kirsten A. Snook
Affiliation:
Department of Pharmaceutical Sciences, Daniel K. Inouye College of Pharmacy, University of Hawai‘i at Hilo, Hilo, Hawai‘i, USA
Argon Steel
Affiliation:
Department of Pharmaceutical Sciences, Daniel K. Inouye College of Pharmacy, University of Hawai‘i at Hilo, Hilo, Hawai‘i, USA
*
Author for correspondence: Susan I. Jarvi, E-mail: [email protected]
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Abstract

Angiostrongylus cantonensis is a parasitic nematode known to infect humans through the ingestion of third stage larvae which can cause inflammation and damage to the central nervous system. Currently, polymerase chain reaction (PCR) is one of the most reliable diagnostic methods for detecting A. cantonensis in humans as well as in gastropod hosts, but requires expensive and specialized equipment. Here, we compare the sensitivity and accuracy of a recombinase polymerase amplification Exo (RPA-EXO) assay, and a recombinase polymerase amplification lateral flow assay (RPA-LFA) with a traditional quantitative PCR (qPCR) assay currently available. The three assays were used to test 35 slugs from Hawai‘i for the presence of A. cantonensis DNA. Consistent results among the three tests were shown in 23/35 samples (65.7%), while 7/35 (20%) were discordant in low infection level samples (<0.01 larvae per mg tissue), and 5/35 (14.3%) were equivocal. To evaluate sensitivity, a partial ITS1 gene was cloned, and serial plasmid dilutions were created ranging from 100 copies μL−1 to ~1 copy μL−1. All three assays consistently detected 50–100 copies μL−1 in triplicate and qPCR was able to detect ~13 copies μL−1 in triplicate. RPA-EXO was able to detect 25 copies μL−1 in triplicate and RPA-LFA was not able to amplify consistently below 50 copies μL−1. Thus, our RPA-EXO and RPA-LFA assays do not appear as sensitive as the current qPCR assay at low DNA concentrations; however, these tests have numerous advantages that may make them useful alternatives to qPCR.

Keywords

Angiostrongylus cantonensislateral flowLFAPCRrecombinase polymeraseRPARPA-EXORPA-LFA
Type
Research Article
Information
Parasitology , Volume 148 , Special Issue 2: 6th International Workshop on Angiostrongylus and Angiostrongyliasis, Hilo, Hawai'i, USA, January, 2020 , February 2021 , pp. 251 - 258
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Copyright
Copyright © The Author(s) 2020. Published by Cambridge University Press

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References

Abingdon Health (2017) What is a lateral flow immunoassay and how does it work? Retrieved from https://www.abingdonhealth.com/contract-services/what-is-a-lateral-flow-immunoassay/.Google Scholar
Alicata, JE (1964) Parasitic infections of man and animals in Hawaii. Hawaii Agricultural Experiment Station Technical Bulletin 61, 1–138.Google Scholar
Alicata, JE (1967) Effect of freezing and boiling on the infectivity of third-stage larvae of Angiostrongylus cantonensis present in land snails and freshwater prawns. Journal of Parasitology 53, 1064–1066.CrossRefGoogle ScholarPubMed
Atkinson, CT, Watcher-Weatherwax, W, Roy, K, Heller, WP and Keith, LM (2017) A Rapid Diagnostic Test and Mobile ‘Lab in a Suitcase’ Platform for Detecting Ceratocystis spp. Responsible for Rapid Ohia Death. Technical Report: Hawaii Cooperative Studies Unit 082.Google Scholar
Barratt, J, Chan, D, Sandaradura, I, Malik, R, Spielman, D, Lee, R, Marriott, D, Harkness, J, Ellis, J and Stark, D (2016). Angiostrongylus cantonensis: a review of its distribution, molecular biology and clinical significance as a human pathogen. Parasitology 143, 1087–1118.CrossRefGoogle ScholarPubMed
Chen, MX, Chen, JX, Chen, SH, Huang, DN, Ai, L and Zhang, RL (2016) Development of lateral 303 flow immunoassay for antigen detection in human Angiostrongylus cantonensis infection. Korean Journal of Parasitology 54, 375–380.CrossRefGoogle Scholar
Clark, K, Karsch-Mizrachi, I, Lipman, DJ, Ostell, J and Sayers, EW (2016) GenBank. Nucleic Acids Research 44, D67–D72.CrossRefGoogle ScholarPubMed
Crannell, ZA, Rohrman, B and Richards-Kortum, R (2014) Equipment-free incubation of recombinase polymerase amplification reactions using body heat. PLoS ONE 9, e112146.CrossRefGoogle ScholarPubMed
Eamsobhana, P, Yong, HS, Mak, JW and Wattanakulpanich, D (1997) Antigenic differences between Parastrongylus cantonensis and Parastrongylus malaysiensis revealed by a monoclonal antibody. International Journal of Infectious Diseases 2, 34–36.CrossRefGoogle Scholar
Eamsobhana, P, Yoolek, A and Punthuprapasa, P (2003) Dot-blot ELISA for the immunological detection of specific antibody to Parastrongylus cantonensis. Journal of Helminthology, 78, 287–291.CrossRefGoogle Scholar
Eamsobhana, P, Tungtrongchitr, A, Wanachiwanawin, D and Yong, HS (2018) Immunochromatographic test for rapid serological diagnosis of human angiostrongyliasis. International Journal of Infectious Diseases 73, 69–71.CrossRefGoogle ScholarPubMed
Eamsobhana, P, Prasartvit, A, Yong, HS, Tungtrongchitr, A and Wanachiwanawin, D (2019) Evaluation of a user-friendly test device (AcQuickDx) for detection of specific antibodies to human angiostrongyliasis. Journal of Food Science & Technology 4, 748–752.Google Scholar
Ghosh, DK, Kokane, SB, Kokane, AD, Warghane, AJ, Motghare, MR, Bhose, S, Sharma, AK and Reddy, MK (2018) Development of a recombinase polymerase based isothermal amplification combined with lateral flow assay (HLB-RPA-LFA) for rapid detection of ‘Candidatus liberibacter asiaticus’. PLoS ONE 13, e0208530.CrossRefGoogle Scholar
Hollingsworth, RG, Kaneta, R, Sullivan, JJ, Bishop, HS, Qvarnstrom, Y, da Silva, AJ and Robinson, DG (2007) Distribution of Parmarion cf. martensi (Pulmonata: Helicarionidae), a new semi-slug pest on Hawai‘i Island, and its potential as a vector for human angiostrongyliasis. Pacific Science 61, 457–467.CrossRefGoogle Scholar
Howe, K, Kaluna, L, Lozano, A, Fischer, BT, Tagami, Y, McHugh, R and Jarvi, S (2019) Water transmission potential of Angiostrongylus cantonensis: larval viability and effectiveness of rainwater catchment sediment filters. PLoS ONE 14, e0209813.CrossRefGoogle ScholarPubMed
Jarvi, SI, Farias, ME, Howe, K, Jacquier, S, Hollingsworth, R and Pitt, W (2012) Quantitative PCR estimates Angiostrongylus cantonensis (rat lungworm) infection levels in semi-slugs (Parmarion martensi). Molecular and Biochemical Parasitology, 185, 174–176.CrossRefGoogle Scholar
Jarvi, SI, Pitt, WC, Farias, MEM, Shiels, L, Severino, M, Howe, K, Jacquier, S, Shiels, AB, Amano, K, Luiz, B, Maher, D, Allison, M, Holtquist, Z and Scheibelhut, N (2015) Detection of Angiostrongylus cantonensis in the blood and peripheral tissues of wild Hawaiian rats (Rattus rattus) by a quantitative PCR (qPCR) assay. PLoS ONE 10, e0123064. doi: 10.1371/journal.pone.0123064CrossRefGoogle ScholarPubMed
Jarvi, SI, Quarta, S, Jacquier, S, Howe, K, Bicakci, D, Dasalla, C, Lovesy, N, Snook, K, McHugh, R and Niebuhr, C (2017) High prevalence of Angiostrongylus cantonensis (rat lungworm) on eastern Hawai‘i Island: a closer look at life cycle traits and patterns of infection in wild rats (Rattus spp.). PLoS ONE 12, e0189458.CrossRefGoogle Scholar
Jarvi, S, Eamsobhana, P, Quarta, S, Howe, K, Jacquier, S, Hanlon, A, Snook, K, McHugh, R, Tman, Z, Miyamura, J, Kramer, K and Myers, K (2020) Estimating human exposure to rat lungworm (Angiostrongylus cantonensis) on Hawai‘i Island: a pilot study. American Journal of Tropical Medicine and Hygiene 102, 69–77.CrossRefGoogle ScholarPubMed
Johnston, DI, Dixon, MC, Elm, JL, Calimlim, PS, Sciulli, RH and Park, SY (2019) Review of cases of angiostrongyliasis in Hawaii, 2007–2017. American Journal of Tropical Medicine and Hygiene 101, 608–616.CrossRefGoogle ScholarPubMed
Kim, JR, Hayes, KA, Yeung, NW and Cowie, RH (2014) Diverse gastropod hosts of Angiostrongylus cantonensis, the rat lungworm, globally and with a focus on the Hawaiian Islands. PLoS ONE 9, e94969. https://doi.org/10.1371/journal.pone.0094969.CrossRefGoogle ScholarPubMed
Kim, JR, Wong, TM, Curry, PA, Yeung, NW, Hayes, KA and Cowie, RH (2019) Modelling the distribution in Hawaii of Angiostrongylus cantonensis (rat lungworm) in its gastropod hosts. Parasitology 146, 42–49.CrossRefGoogle ScholarPubMed
Koczula, KM and Gallotta, A (2016). Lateral flow assays. Essays in Biochemistry 60, 111–120.Google ScholarPubMed
Kumar, S, Stecher, G and Tamura, K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33, 1870–1874.CrossRefGoogle ScholarPubMed
Liu, EW, Schwarts, BS, Hysmith, ND, DeVinco, JP, Larson, DT, Maves, RC, Palazzi, DL, Meyer, C, Custido, HT, Braza, MM, Hammond, RA, Rao, S, Qvarnstrom, Y, Yabsley, MJ, Bradbury, RS and Montgomery, SP (2018) Rat lungworm infection associated with central nervous system disease – eight U.S. states, January 2011–January 2017. MMWR. Morbidity and Mortality Weekly Report 67, 825–828.CrossRefGoogle ScholarPubMed
Niebuhr, CN, Jarvi, SI, Kaluna, L, Fischer, BLT, Deane, AR, Leinbach, IL and Siers, SR (2020) Occurrence of rat lungworm (Angiostrongylus cantonensis) in invasive Coqui frogs (Eleutherodactylus coqui) and other hosts in Hawaii, USA. Journal of Wildlife Diseases 56, 203–207.CrossRefGoogle ScholarPubMed
Park, BH, Oh, SJ, Jung, JH, Choi, G, Seo, JH, Kim, H, Lee, EY and Seo, TS (2017) An integrated rotary microfluidic system with DNA extraction, loop-mediated isothermal amplification, and lateral flow strip based detection for point-of-care pathogen diagnostics. Biosensors and Bioelectronics 91, 334–340.CrossRefGoogle ScholarPubMed
Piepenburg, O, Williams, CH, Stemple, DL and Armes, NA (2006) DNA detection using recombination proteins. PLoS Biology 4, e204.CrossRefGoogle ScholarPubMed
Qvarnstrom, Y, da Silva, AC, Teem, JL, Hollingsworth, R, Bishop, H, Graeff-Teixeira, C and da Silva, AJ (2010) Improved molecular detection of Angiostrongylus cantonensis in mollusks and other environmental samples with a species-specific internal transcribed spacer 1-based TaqMan assay. Applied Environmental Microbiology 76, 5287–5289.CrossRefGoogle ScholarPubMed
Qvarnstrom, Y, Xayavong, M, da Silva, A, Park, S, Whelen, A, Calimlim, P, Sciulli, R, Honda, S, Higa, K, Kitsutani, P, Chea, N, Heng, S, Johnson, S, Graeff-Texeira, C, Fos, L and da Silva, A (2016) Real-time polymerase chain reaction detection of Angiostrongylus cantonensis DNA in cerebrospinal fluid from patients with eosinophilic meningitis. American Journal of Tropical Medicine and Hygiene 94, 176–181.CrossRefGoogle ScholarPubMed
Wang, Q, Lai, D, Zhu, X, Che, X and Lun, Z (2008) Human angiostrongyliasis. The Lancet Infectious Diseases 8, 621–630.CrossRefGoogle ScholarPubMed
Wang, QP, Wu, ZD, Wei, J, Owen, RL and Lun, ZR (2012). Human Angiostrongylus cantonensis: an update. European Journal of Clinical Microbiology & Infectious Disease 31, 389–395.CrossRefGoogle ScholarPubMed
Zou, Y, Mason, MG, Wang, Y, Wee, E, Turni, C, Blackall, PJ, Trau, M and Botella, JR (2017) Nucleic acid purification from plants, animals and microbes in under 30 seconds. PLoS Biology 15, e2003916.CrossRefGoogle ScholarPubMed