Comparison of different molecular protocols for the detection of Uncinaria stenocephala infection in dogs.

Autor: Illiano S; Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Via Delpino 1, Naples 80137, Italy. Electronic address: sergio.illiano@unina.it., Ciuca L; Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Via Delpino 1, Naples 80137, Italy. Electronic address: lavinia.ciuca@unina.it., Bosco A; Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Via Delpino 1, Naples 80137, Italy. Electronic address: antonio.bosco@unina.it., Rinaldi L; Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Via Delpino 1, Naples 80137, Italy. Electronic address: laura.rinaldi@unina.it., Maurelli MP; Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Via Delpino 1, Naples 80137, Italy. Electronic address: mariapaola.maurelli@unina.it.
Jazyk: angličtina
Zdroj: Veterinary parasitology [Vet Parasitol] 2024 Aug; Vol. 330, pp. 110249. Date of Electronic Publication: 2024 Jul 03.
DOI: 10.1016/j.vetpar.2024.110249
Abstrakt: The present study aims to assess the performance of different molecular targets using various matrices of samples for the detection of Uncinaria stenocephala (US) in hookworm infected dogs. To this end, the DNA extraction was performed on the following matrices of samples: (i) larvae of US obtained from experimentally infected dogs with US with different larvae counts per microliter (µl); (ii) pure US eggs suspension in distilled water with different egg counts per µl; (iii) spiked dog fecal samples with different US eggs per gram (EPG) of feces; (iv) feces from dogs naturally infected with hookworm eggs; (v) fecal suspension with hookworm eggs recovered from the FLOTAC apparatus. All the samples were tested with four different PCR protocols targeting specific regions for the detection of both hookworms US and AC as follows: Protocol A (ITS1, 5.8 S, ITS2) and Protocol B (18 S) for the detection of both species, Protocol C (ITS1) for the detection of AC and Protocol D (ITS1) for the detection of US. The best results were obtained with DNA extracted from US larvae matrix obtained from experimentally infected dogs, showing a detection limit of 3.5 larvae/ml for the protocols A, B and D. A moderate correlation was found between the FLOTAC technique and PCR protocols B and D with respect to fecal samples from dogs naturally infected with hookworms. Indeed, PCR protocols B (18 S) and D (ITS1) gave the best results for feces and fecal suspension from naturally infected dogs. However, all the PCR protocols used showed lower sensitivity than FLOTAC technique. Perhaps, isolating US eggs in advance could help to obtain better quality and quantity of DNA, avoiding some notable factors such as inhibitors present in faecal samples. However, a further study is needed to evaluate and standardise a protocol for the recovery of parasitic elements, that could be applied prior to DNA extraction. Therefore, this could lead to a better amplification of US eggs DNA. In conclusion, our results showed that the type of sample (sample-matrix) used for the DNA extraction samples is crucial, as this affects the diagnostic sensitivity of the technique.
Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Lavinia Ciuca reports was provided by University of Naples Federico II. Lavinia Ciuca reports a relationship with University of Naples Federico II that includes: employment. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper
(Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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