Home About Browse Search
Svenska


A novel duplex ddPCR assay for detection and differential diagnosis of Ascaridia galli and Heterakis gallinarum eggs from chickens feces

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]

[img] PDF
1MB

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
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:
Type of IDID
DOI10.1016/j.vetpar.2021.109499
Web of Science (WoS)000674663800018
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

Repository Staff Only: item control page

Downloads

Downloads per year (since September 2012)

View more statistics

Downloads
Hits