- Title
- Applicability of LAMP as a field diagnostic test for haemonchus contortus and fasciola hepatica infection
- Creator
- Bari, Tanjina
- Date
- 2021
- Type
- Text; Thesis; PhD
- Identifier
- http://researchonline.federation.edu.au/vital/access/HandleResolver/1959.17/178836
- Identifier
- vital:15461
- Abstract
- Gastrointestinal parasites Haemonchus contortus and Fasciola hepatica are major impediments to livestock production worldwide. Faecal egg counts remain the most commonly used and widely accepted diagnostic tool for these parasites; however, there is a need for improved, field-applicable diagnostics. In this study, loop-mediated isothermal amplification (LAMP) was optimised and evaluated for the detection of H. contortus (in sheep) and F. hepatica (in cattle) infection. LAMP assays were optimised to enable visual detection using calcein dye. DNA extraction techniques were developed that have the potential for on-farm application. Faeces suspended in water, heated, then centrifuged, with two cheap and stable chemicals, enabled detection of H. contortus at clinically relevant infection burdens. For F. hepatica, a faeces-water suspension was sieved to remove particulate matter, then physical disruption (bead-beating) was applied. LAMP was conducted under laboratory conditions and in the field; compared to FEC (the most commonly used diagnostic for the target parasites) and PCR. LAMP was conducted using three incubation methods: a commercially available thermocycler; a field-friendly low-cost portable styrofoam esky; and a dedicated field applicable LAMP incubator. The general trend was for LAMP to have high sensitivity but only moderate specificity when compared to FEC. However, the use of PCR (both pathogens) and a highly sensitive amended FEC (F. hepatica only) suggested that the apparent low specificity was the result of LAMP being able to detect low-level parasite infection when conventional FEC could not. A LAMP assay paired with a potentially field-applicable DNA extraction was able to adequately detect haemonchosis at ‘clinically relevant’ parasite burdens in a laboratory study. A field study for the detection of F. hepatica demonstrated the potential utility of LAMP on-farm. The studies conducted in this thesis demonstrate the potential of LAMP for parasitic disease diagnosis in agriculture.; Doctor of Philosophy
- Publisher
- Federation University Australia
- Rights
- All metadata describing materials held in, or linked to, the repository is freely available under a CC0 licence
- Rights
- Copyright Tanjina Bari
- Rights
- Open Access
- Subject
- Haemonchus contortus; Fasciola hepatica; Loop mediated isothermal amplification
- Full Text
- Thesis Supervisor
- Greenhill, Andrew
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