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Viruses 2018, 10(5), 247; https://doi.org/10.3390/v10050247

Multiplex Detection of Aspergillus fumigatus Mycoviruses

1
Department of Molecular Biology and Genetics, Ahi Evran University, Kırşehir 40100, Turkey
2
Department of Life Sciences, Faculty of Natural Sciences, Imperial College London, London SW7 2AZ, UK
3
Department of Biological and Environmental Sciences, University of Hertfordshire, Hatfield AL10 9AB, UK
*
Author to whom correspondence should be addressed.
Received: 5 April 2018 / Revised: 5 May 2018 / Accepted: 6 May 2018 / Published: 8 May 2018
(This article belongs to the Special Issue Mycoviruses)
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Abstract

Mycoviruses are viruses that naturally infect and replicate in fungi. They are widespread in all major fungal groups including plant and animal pathogenic fungi. Several dsRNA mycoviruses have been reported in Aspergillus fumigatus. Multiplex polymerase chain reaction (PCR) amplification is a version of PCR that enables amplification of different targets simultaneously. This technique has been widely used for detection and differentiation of viruses especially plant viruses such as those which infect tobacco, potato and garlic. For rapid detection, multiplex RT-PCR was developed to screen new isolates for the presence of A. fumigatus mycoviruses. Aspergillus fumigatus chrysovirus (AfuCV), Aspergillus fumigatus partitivirus (AfuPV-1), and Aspergillus fumigatus tetramycovirus-1 (AfuTmV-1) dsRNAs were amplified in separate reactions using a mixture of multiplex primer pairs. It was demonstrated that in the presence of a single infection, primer pair mixtures only amplify the corresponding single virus infection. Mixed infections using dual or triple combinations of dsRNA viruses were also amplified simultaneously using multiplex RT-PCR. Up until now, methods for the rapid detection of Aspergillus mycoviruses have been restricted to small scale dsRNA extraction approaches which are laborious and for large numbers of samples not as sensitive as RT-PCR. The multiplex RT-PCR assay developed here will be useful for studies on determining the incidence of A. fumigatus mycoviruses. This is the first report on multiplex detection of A. fumigatus mycoviruses. View Full-Text
Keywords: dsRNA mycoviruses; multiplex PCR; Aspergillus fumigatus chrysovirus; Aspergillus fumigatus partitivirus-1; Aspergillus fumigatus tetramycovirus-1. dsRNA mycoviruses; multiplex PCR; Aspergillus fumigatus chrysovirus; Aspergillus fumigatus partitivirus-1; Aspergillus fumigatus tetramycovirus-1.
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Özkan-Kotiloğlu, S.; Coutts, R.H.A. Multiplex Detection of Aspergillus fumigatus Mycoviruses. Viruses 2018, 10, 247.

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