Real-time PCR assay for screening Pneumocystis in free-living wild squirrels and river rats in Italy
| Autor: | Graziana Da Rold, Gioia Capelli, Michela Corrò, Elena Porcellato, Claudia Cafarchia, Patrizia Danesi, Antonio Frangipane di Regalbono, Christian Falcaro, Silvia Ravagnan, Wieland Meyer, Roberta Iatta |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Callosciurus finlaysonii Myocastor coypus Pneumocystis free-living wild rodents rRNA real-time PCR Protein subunit 030106 microbiology Rodentia Biology Real-Time Polymerase Chain Reaction Rodent Diseases 03 medical and health sciences chemistry.chemical_compound Prevalence Animals Full Scientific Reports DNA Fungal Lung Pathogen Phylogeny General Veterinary Phylogenetic tree Sequence Analysis RNA Sciuridae Ribosomal RNA Amplicon Molecular biology Pneumocystis Infections 030104 developmental biology Real-time polymerase chain reaction Italy chemistry RNA Ribosomal Introduced Species Nested polymerase chain reaction DNA |
| Popis: | We used a real-time PCR (rtPCR) targeting a 150-bp amplicon of the mitochondrial small subunit of ribosomal RNA (mtSSU rRNA) to screen for Pneumocystis DNA in lungs of wild squirrels ( Callosciurus finlaysonii, n = 85) and river rats ( Myocastor coypus, n = 43) in Italy. The rtPCR revealed Pneumocystis DNA in 20 of 85 (24%) squirrels and in 35 of 43 (81%) river rats, and was more sensitive than a nested PCR that targets a portion of the mtSSU rRNA and the mitochondrial large subunit of rRNA (mtLSU rRNA). Phylogenetic analysis based on mtSSU rRNA and mtLSU rRNA sequences showed distinct Pneumocystis sequence types in these rodents. The rtPCR assay should be reliable for screening large populations for this potential pathogen, thereby allowing cost-effective monitoring of the disease in wild animals. |
| Databáze: | OpenAIRE |
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