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Assessment of three DNA extraction kits for the absolute quantification of strongyle nematode eggs in faecal samples

Högberg, Niclas and Baltrusis, Paulius and Enweji, Nizar and Höglund, Johan (2022). Assessment of three DNA extraction kits for the absolute quantification of strongyle nematode eggs in faecal samples. Acta Veterinaria Scandinavica. 64 :1 , 5
[Research article]

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Abstract

Background Haemonchus contortus is one of the most pathogenic gastrointestinal nematodes of small ruminants. The current diagnostic approach for the detection of this species relies on coproscopic methods, which both have low sensitivity and are time consuming. Methods employing detection through DNA amplification, such as droplet digital polymerase chain reaction (ddPCR), offer an advantageous approach to the diagnosis of H. contortus. However, DNA extraction protocols need to be constantly updated for the optimal retrieval of diagnostically usable template. Here, we describe the evaluation of three genomic DNA extraction kits for the detection and quantification of H. contortus ITS2 amplicon DNA from faecal samples, using droplet digital PCR. Results DNA samples, extracted from faecal material with the Nucleospin DNA Stool kit, produced the highest amounts of ITS2 amplicon copies and had the lowest coefficient of variation across different dilutions and sample types (fresh or frozen) out of the tested kits (Nucleospin DNA Stool, E.Z.N.A.(R) Stool DNA Kit and QIAamp Fast DNA Stool Mini Kit). Furthermore, the protocol of this kit has the fewest number of steps and the price of DNA extraction per sample is reasonable (2.77 euro). Conclusions The Nucleospin DNA Stool kit is an attractive option for the detection and quantification of H. contortus DNA in faecal samples of small ruminants in a diagnostic setting.

Authors/Creators:Högberg, Niclas and Baltrusis, Paulius and Enweji, Nizar and Höglund, Johan
Title:Assessment of three DNA extraction kits for the absolute quantification of strongyle nematode eggs in faecal samples
Series Name/Journal:Acta Veterinaria Scandinavica
Year of publishing :2022
Volume:64
Number:1
Article number:5
Number of Pages:9
Publisher:BMC
ISSN:0044-605X
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 > Pathobiology
Keywords:Diagnostics, DNA isolation, Haemonchus, Nematode eggs, Parasite, PCR, Sheep
URN:NBN:urn:nbn:se:slu:epsilon-p-116287
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-p-116287
Additional ID:
Type of IDID
DOI10.1186/s13028-022-00624-3
Web of Science (WoS)000753379000002
ID Code:27259
Faculty:VH - Faculty of Veterinary Medicine and Animal Science
Department:(VH) > Department of Biomedical Science and Veterinary Public Health
Deposited By: SLUpub Connector
Deposited On:07 Mar 2022 10:25
Metadata Last Modified:07 Mar 2022 10:31

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