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A Sarcoptes scabiei specific isothermal amplification assay for detection of this important ectoparasite of wombats and other animals
Citation
Fraser, TA and Carver, S and Martin, AM and Mounsey, K and Polkinghorne, A and Jelocnik, M, A Sarcoptes scabiei specific isothermal amplification assay for detection of this important ectoparasite of wombats and other animals, PeerJ, 6 pp. e5291. ISSN 2167-8359 (2018) [Refereed Article]
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Copyright Statement
2018 Fraser et al. This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0)License, (https://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.
Abstract
Background
The globally distributed epidermal ectoparasite, Sarcoptes scabiei, is a serious health and welfare burden to at-risk human and animal populations. Rapid and sensitive detection of S. scabiei infestation is critical for intervention strategies. While direct microscopy of skin scrapings is a widely utilised diagnostic method, it has low sensitivity. PCR, alternatively, has been shown to readily detect mite DNA even in microscopy-negative skin scrapings. However, a limitation to the latter method is the requirements for specialised equipment and reagents. Such resources may not be readily available in regional or remote clinical settings and are an important consideration in diagnosis of this parasitic disease.
Methodology
A Loop Mediated Isothermal Amplification (LAMP) assay targeting the ITS-2 gene for S. scabiei was developed and evaluated on clinical samples from various hosts, previously screened with conventional S. scabies-specific PCR. Species specificity of the newly developed LAMP assay was tested against a range of DNA samples from other arthropods. The LAMP assays were performed on a real-time fluorometer as well as thermal cycler to evaluate an end-point of detection. Using skin scrapings, a rapid sample processing method was assessed to eliminate extensive processing times involved with DNA extractions prior to diagnostic assays, including LAMP.
Results
The S. scabiei LAMP assay was demonstrated to be species-specific and able to detect DNA extracted from a single mite within a skin scraping in under 30 minutes. Application of this assay to DNA extracts from skin scrapings taken from a range of hosts revealed 92.3% congruence (with 92.50% specificity and 100% sensitivity) to the conventional PCR detection of S. scabiei. Preliminary results have indicated that diagnostic outcome from rapidly processed dry skin scrapings using our newly developed LAMP is possible in approximately 40 minutes.
Discussion
We have developed a novel, rapid and robust molecular assay for detecting S. scabiei infesting humans and animals. Based on these findings, we anticipate that this assay will serve an important role as an ancillary diagnostic tool at the point-of-care, complementing existing diagnostic protocols for S. scabiei.
Item Details
| Item Type: | Refereed Article |
|---|---|
| Keywords: | epidemiology, LAMP, diagnostics, sarcoptic mange, skin scraping, PCR, one health, Australian wildlife, Sarcoptes scabiei, wombats |
| Research Division: | Agricultural, Veterinary and Food Sciences |
| Research Group: | Veterinary sciences |
| Research Field: | Veterinary parasitology |
| Objective Division: | Environmental Management |
| Objective Group: | Terrestrial systems and management |
| Objective Field: | Control of pests, diseases and exotic species in terrestrial environments |
| UTAS Author: | Fraser, TA (Ms Tamieka Fraser) |
| UTAS Author: | Carver, S (Associate Professor Scott Carver) |
| UTAS Author: | Martin, AM (Ms Alynn Martin) |
| ID Code: | 149600 |
| Year Published: | 2018 |
| Web of Science® Times Cited: | 10 |
| Deposited By: | Zoology |
| Deposited On: | 2022-04-05 |
| Last Modified: | 2022-05-24 |
| Downloads: | 6 View Download Statistics |
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