Tarbiat, Behdad and Enweji, Nizar and Baltrusis, Paulius and Halvarsson, Peter and Osterman-Lind, Eva and Jansson, Desirée and Höglund, Johan
(2021).
A novel duplex ddPCR assay for detection and differential diagnosis of Ascaridia galli and Heterakis gallinarum eggs from chickens feces.
Veterinary Parasitology. 296
, 109499
[Research article]
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Abstract
Since the EU ban on battery cages, many studies have listed Ascaridia galli and Heterakis gallinarum as the most common roundworms in the European laying hen population. A complicating factor is that the eggs of these parasites are almost identical. Thus, lack of molecular diagnostic approaches has driven epidemiological studies to take on necropsy for species discrimination, which is labor and cost intensive. Here, we describe a novel diagnostic tool based on droplet digital PCR for simultaneous identification and absolute quantification of the eggs of both of these ascarids in chickens' droppings using two different genus-specific primer-probe sets targeting the second internal transcribed spacer region (ITS-2) in the nuclear ribosomal (rRNA) gene array. No cross-reaction was observed when different combinations of DNA and species-specific primers and probes were tested. The lowest obtained frequency threshold for the detection of H. gallinarum in the presence of a constant A. galli DNA concentration was determined to be 0.8 %. After validation, we used the assay to analyze field samples collected from several Swedish laying hen farms. Out of 134 samples, 86 (64 %) were positive for A. galli while 11 (8.3 %) samples were positive for H. gallinarum. These samples were initially analyzed with flotation technique for detection of ascarid eggs. The results of the Cohen's kappa indicated substantial agreement (85.8 %) between the two tests. In conclusion, we have validated a novel molecular-based diagnostic tool for quantification and differentiation between intestinal parasites of major importance in chickens with high precision. Although this study focuses on identification of parasites of laying hens, the findings may well have a bearing on all types of chicken production systems. The present study lays the groundwork for future research into epidemiology of these two important chicken parasite species.
Authors/Creators: | Tarbiat, Behdad and Enweji, Nizar and Baltrusis, Paulius and Halvarsson, Peter and Osterman-Lind, Eva and Jansson, Desirée and Höglund, Johan | ||||||
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Title: | A novel duplex ddPCR assay for detection and differential diagnosis of Ascaridia galli and Heterakis gallinarum eggs from chickens feces | ||||||
Series Name/Journal: | Veterinary Parasitology | ||||||
Year of publishing : | 2021 | ||||||
Volume: | 296 | ||||||
Article number: | 109499 | ||||||
Number of Pages: | 7 | ||||||
Publisher: | ELSEVIER | ||||||
ISSN: | 0304-4017 | ||||||
Language: | English | ||||||
Publication Type: | Research article | ||||||
Article category: | Scientific peer reviewed | ||||||
Version: | Published version | ||||||
Copyright: | Creative Commons: Attribution 4.0 | ||||||
Full Text Status: | Public | ||||||
Subjects: | (A) Swedish standard research categories 2011 > 4 Agricultural Sciences > 403 Veterinary Science > Clinical Science | ||||||
Keywords: | Roundworm, Laying hens, Molecular assay, Poultry, Parasites, Diagnostic, Prevalence | ||||||
URN:NBN: | urn:nbn:se:slu:epsilon-p-113133 | ||||||
Permanent URL: | http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-p-113133 | ||||||
Additional ID: |
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ID Code: | 25073 | ||||||
Faculty: | VH - Faculty of Veterinary Medicine and Animal Science | ||||||
Department: | (VH) > Department of Biomedical Science and Veterinary Public Health (VH) > Dept. of Clinical Sciences | ||||||
Deposited By: | SLUpub Connector | ||||||
Deposited On: | 27 Aug 2021 09:25 | ||||||
Metadata Last Modified: | 27 Aug 2021 09:31 |
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