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Molecular diagnosis of Pneumocystis pneumonia in dogs
Journal article   Peer reviewed

Molecular diagnosis of Pneumocystis pneumonia in dogs

P. Danesi, S. Ravagnan, L.R. Johnson, T. Furlanello, A. Milani, P. Martin, S. Boyd, M. Best, B. Galgut, P. Irwin, …
Medical Mycology, Vol.55(8), pp.828-842
2017
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Abstract

Pneumocystis pneumonia (PCP) is a life-threatening fungal disease that can occur in dogs. The aim of this study was to provide a preliminary genetic characterisation of Pneumocystis carinii f. sp. 'canis' (P. canis) in dogs and thereby develop a reliable molecular protocol to definitively diagnose canine PCP. We investigated P. canis in a variety of lung specimens from dogs with confirmed or strongly suspected PCP (Group 1, n = 16), dogs with non-PCP lower respiratory tract problems (Group 2, n = 65) and dogs not suspected of having PCP or other lower respiratory diseases (Group 3, n = 11). Presence of Pneumocystis DNA was determined by nested PCR of the large and small mitochondrial subunit rRNA loci and by a real-time quantitative polymerase chain reaction (qPCR) assay developed using a new set of primers. Molecular results were correlated with the presence of Pneumocystis morphotypes detected in cytological/histological prepa rations. Pneumocystis DNA was amplified from 13/16 PCP-suspected dogs (Group 1) and from 4/76 dogs of control Groups 2 and 3 (combined). The latter four dogs were thought to have been colonized by P. canis. Comparison of C-T values in 'infected' versus 'colonized' dogs was consistent with this notion, with a distinct difference in molecular burden between groups (C-T <= 26 versus C-T range (26 < C-T < 35), respectively). Phylogenetic analyses showed that P. canis is specifically 'canine' associated, being separated from other mammalian Pneumocystis species, thereby confirming the accuracy of qPCR amplicon for Pneumocystis in dogs. Using qPCR, Pneumocystis DNA can be detected in specimens from the respiratory tract and a C-T value can be interpreted to distinguish infection versus colonization.

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Collaboration types
Domestic collaboration
International collaboration
Citation topics
1 Clinical & Life Sciences
1.66 HIV
1.66.1769 Pneumocystis-Carinii Pneumonia
Web Of Science research areas
Infectious Diseases
Mycology
Veterinary Sciences
ESI research areas
Plant & Animal Science
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