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001-es BibID:BIBFORM112724
035-os BibID:(cikkazonosító)109974 (WoS)001035781600001 (Scopus)85163192208
Első szerző:Khangembam, Rojesh
Cím:Preliminary Results of the Recombinase Polymerase Amplification Technique for the Detection of Haemonchus contortus from Hungarian Field Samples / Rojesh Khangembam; Nóra Vass; Alison Morrison; Lynsey A Melville; Alistair Antonopoulos; Levente Czeglédi
Dátum:2023
ISSN:0304-4017
Megjegyzések:Haemonchus contortus is a parasitic nematode of small ruminants responsible for significant economic losses and animal health concerns globally. Detection of gastrointestinal nematode (GIN) infection in veterinary practice typically relies on microscopy-based methods such as the faecal egg count and morphological identification of larval culture. However, mixed co-infections are common and species-specific identification is typically time-consuming and expertise-intensive. Compounded by increasing anthelmintic resistance, there is an urgent need to implement the molecular diagnosis of GIN in the livestock industry, preferably in field settings. Advances in isothermal amplification techniques including recombinase polymerase amplification (RPA) assays could improve this. Yet, constraints in RPA kit availability and amplicon detection systems limit the use of this technology in point of care settings. In this study, we present an early-stage, proof-of-concept demonstration of RPA targeting the internal transcribed spacer (ITS2) region of H. contortus. Having tested against eight closely related nematodes and also against five farm isolates in Eastern Hungary, preliminary results derived from a comparative analysis of 3 primer sets showed the assay detects H. contortus DNA and has a limit of detection of 10-5 ng/?l. We also tested an end-result naked eye detection system using various DNA binding dyes, of which EvaGreen® dye was successful for a qualitative RPA detection that could be adaptable at farm sites.
Tárgyszavak:Agrártudományok Állatorvosi tudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Haemonchus
RPA
isothermal amplification
point of care
diagnostic
Megjelenés:Veterinary Parasitology. - 320 (2023), p. 1-20. -
További szerzők:Vass Nóra (1983-) (állatorvos) Morrison, Alison Melville, Lynsey A Antonopoulos, Alistair Czeglédi Levente (1977-) (agrármérnök)
Pályázati támogatás:EFOP-3.6.3-VEKOP-16-2017-00008
EFOP-VEKOP
Internet cím:DOI
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