title: Ubiquiter circovirus sequences raise challenges in laboratory diagnosis: the case of honey bee and bee mite, reptiles, and free living amoebae.
abstract: Circoviruses of pigs and birds are established pathogens, however, the exact role of other, recently described circoviruses and circovirus-like viruses remains to be elucidated. The aim of this study was the detection of circoviruses in neglected host species, including honey bees, exotic reptiles and free-living amoebae by widely used broad-spectrum polymerase chain reaction (PCR) assays specific for the replication initiation protein coding gene of these viruses. The majority of sequences obtained from honey bees were highly similar to canine and porcine circoviruses, or, were distantly related to dragonfly cycloviruses. Other rep sequences detected in some honey bees, reptiles and amoebae showed similarities to various rep sequences deposited in the GenBank. Back-to-back PCR primers designed for the amplification of whole viral genomes failed to work that suggested the existence of integrated rep-like elements in many samples. Rolling circle amplification and exonuclease treatment confirmed the absence of small circular DNA genomes in the specimens analysed. In case of honey bees Varroa mite DNA contamination might be a source of the identified endogenous rep-like elements. The reptile and amoebae rep-like sequences were nearly identical with each other and with sequences detected in chimpanzee feces raising the possibility that detection of novel or unusual rep-like elements in some host species might originate from the microbial community of the host. Our results indicate that attention is needed when broad-spectrum rep gene specific polymerase chain reaction is chosen for laboratory diagnosis of circovirus infections.
authors.affiliation: Hungarian Academy of Sciences Institute for Veterinary Medical Research, Centre for Agricultural Research Budapest Hungary.
authors.forename: Szilvia
authors.initials: S
authors.lastname: Marton
authors.affiliation: Hungarian Academy of Sciences Institute for Veterinary Medical Research, Centre for Agricultural Research Budapest Hungary.
authors.forename: Katalin
authors.initials: K
authors.lastname: Ihász
authors.affiliation: Hungarian Defence Forces Military Medical Centre Budapest Hungary.
authors.forename: György
authors.initials: G
authors.lastname: Lengyel
authors.affiliation: Hungarian Academy of Sciences Institute for Veterinary Medical Research, Centre for Agricultural Research Budapest Hungary.
authors.forename: Szilvia L
authors.initials: SL
authors.lastname: Farkas
authors.affiliation: National Food Chain Safety Office Veterinary Diagnostic Directorate Budapest Hungary.
authors.forename: Ádám
authors.initials: Á
authors.lastname: Dán
authors.affiliation: National Food Chain Safety Office Veterinary Diagnostic Directorate Budapest Hungary.
authors.forename: Petra
authors.initials: P
authors.lastname: Paulus
authors.affiliation: Hungarian Academy of Sciences Institute for Veterinary Medical Research, Centre for Agricultural Research Budapest Hungary.
authors.forename: Krisztián
authors.initials: K
authors.lastname: Bányai
authors.affiliation: Hungarian Academy of Sciences Institute for Veterinary Medical Research, Centre for Agricultural Research Budapest Hungary.
authors.forename: Enikő
authors.initials: E
authors.lastname: Fehér
mesh_terms: D000656:Amoeba / Q000821:virology*; D000818:Animals; D001483:Base Sequence; D001516:Bees / Q000469:parasitology / Q000821:virology*; D018173:Circoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D017925:Circovirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D019411:Clinical Laboratory Techniques; D016679:Genome, Viral / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D012104:Reptiles / Q000821:virology*; D017422:Sequence Analysis, DNA / Q000662:veterinary; D057077:Varroidae / Q000821:virology*
keywords: circovirus; integrated; rep; sequencing