Comparative study of a broad qPCR panel and centrifugal flotation for detection of gastrointestinal parasites in fecal samples from dogs and cats in the United States

Autor: Christian M. Leutenegger, Cecilia E. Lozoya, Jeffrey Tereski, Jan Andrews, Kelly D. Mitchell, Cathy Meeks, Jennifer L. Willcox, Gregory Freeman, Holly L. Richmond, Christian Savard, Michelle D. Evason
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: Parasites & Vectors, Vol 16, Iss 1, Pp 1-10 (2023)
Druh dokumentu: article
ISSN: 1756-3305
DOI: 10.1186/s13071-023-05904-z
Popis: Abstract Background For decades, zinc sulfate centrifugal fecal flotation microscopy (ZCF) has been the mainstay technique for gastrointestinal (GI) parasite screening at veterinary clinics and laboratories. Elsewhere, PCR has replaced microscopy because of generally increased sensitivity and detection capabilities; however, until recently it has been unavailable commercially. Therefore, the primary aim of this study was to compare the performance of real-time PCR (qPCR) and ZCF for fecal parasite screening. Secondary aims included further characterization of markers for hookworm treatment resistance and Giardia spp. assemblages with zoonotic potential and qPCR optimization. Methods A convenience sampling of 931 canine/feline fecal samples submitted to a veterinary reference laboratory for routine ZCF from the Northeast US (11/2022) was subsequently evaluated by a broad qPCR panel following retention release. Detection frequency and agreement (kappa statistics) were evaluated between ZCF and qPCR for seven GI parasites [hookworm/(Ancylostoma spp.), roundworm/(Toxocara spp.), whipworm/(Trichuris spp.), Giardia duodenalis, Cystoisospora spp., Toxoplasma gondii, and Tritrichomonas blagburni] and detections per sample. Total detection frequencies were compared using a paired t-test; positive sample and co-infection frequencies were compared using Pearson’s chi-squared test (p ≤ 0.05 significant) and qPCR frequency for hookworm benzimidazole (BZ) resistance (F167Y) and zoonotic Giardia spp. assemblage markers calculated. Confirmatory testing, characterization, and qPCR optimization were carried out with Sanger sequencing. Results qPCR detected a significantly higher overall parasite frequency (n = 679) compared to ZCF (n = 437) [p =
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