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Article

Detection of Helminth Ova in Wastewater Using Recombinase Polymerase Amplification Coupled to Lateral Flow Strips

1
Centre for Environmental Sustainability and Remediation, School of Science, RMIT University, Bundoora West, VIC 3083, Australia
2
South East Water, Frankston, VIC 3199, Australia
3
Melbourne Water, Docklands, VIC 3008, Australia
*
Author to whom correspondence should be addressed.
Water 2020, 12(3), 691; https://doi.org/10.3390/w12030691
Received: 3 February 2020 / Revised: 26 February 2020 / Accepted: 2 March 2020 / Published: 3 March 2020
Ascaris lumbricoides is a major soil-transmitted helminth that is highly infective to humans. The ova of A. lumbricoides are able to survive wastewater treatment, thus making it an indicator organism for effective water treatment and sanitation. Hence, Ascaris ova must be removed from wastewater matrices for the safe use of recycled water. Current microscopic techniques for identification and enumeration of Ascaris ova are laborious and cumbersome. Polymerase chain reaction (PCR)-based techniques are sensitive and specific, however, major constraints lie in having to transport samples to a centralised laboratory, the requirement for sophisticated instrumentation and skilled personnel. To address this issue, a rapid, highly specific, sensitive, and affordable method for the detection of helminth ova was developed utilising recombinase polymerase amplification (RPA) coupled with lateral flow (LF) strips. In this study, Ascaris suum ova were used to demonstrate the potential use of the RPA-LF assay. The method was faster (< 30 min) with optimal temperature at 37 °C and greater sensitivity than PCR-based approaches with detection as low as 2 femtograms of DNA. Furthermore, ova from two different helminth genera were able to be detected as a multiplex assay using a single lateral flow strip, which could significantly reduce the time and the cost of helminth identification. The RPA-LF system represents an accurate, rapid, and cost-effective technology that could replace the existing detection methods, which are technically challenged and not ideal for on-site detection in wastewater treatment plants. View Full-Text
Keywords: Ascaris; lateral flow; point-of-care; recombinase polymerase amplification; soil-transmitted helminths; wastewater Ascaris; lateral flow; point-of-care; recombinase polymerase amplification; soil-transmitted helminths; wastewater
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MDPI and ACS Style

Ravindran, V.B.; Khallaf, B.; Surapaneni, A.; Crosbie, N.D.; Soni, S.K.; Ball, A.S. Detection of Helminth Ova in Wastewater Using Recombinase Polymerase Amplification Coupled to Lateral Flow Strips. Water 2020, 12, 691. https://doi.org/10.3390/w12030691

AMA Style

Ravindran VB, Khallaf B, Surapaneni A, Crosbie ND, Soni SK, Ball AS. Detection of Helminth Ova in Wastewater Using Recombinase Polymerase Amplification Coupled to Lateral Flow Strips. Water. 2020; 12(3):691. https://doi.org/10.3390/w12030691

Chicago/Turabian Style

Ravindran, Vivek B., Basma Khallaf, Aravind Surapaneni, Nicholas D. Crosbie, Sarvesh K. Soni, and Andrew S. Ball 2020. "Detection of Helminth Ova in Wastewater Using Recombinase Polymerase Amplification Coupled to Lateral Flow Strips" Water 12, no. 3: 691. https://doi.org/10.3390/w12030691

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