Open AccessSacbrood Virus of the Honeybee ( Apis mellifera ): Rapid Identification and Phylogenetic Analysis Using Reverse Transcription-PCR
Author(s) -
Elvira Grabensteiner,
W. Ritter,
Michael J. Carter,
Sean Davison,
Hermann Pechhacker,
Jolanta Kolodziejek,
Otto Boecking,
Irmgard Derakhshifar,
Rudolf Moosbeckhofer,
Elisabeth Licek,
Norbert Nowotny
Publication year - 2001
Publication title -
clinical and diagnostic laboratory immunology
Language(s) - English
Resource type - Journals
eISSN - 1098-6588
pISSN - 1071-412X
DOI - 10.1128/cdli.8.1.93-104.2001
Subject(s) - biology , phylogenetic tree , virology , virus , genome , reverse transcription polymerase chain reaction , genetics , gene , gene expression
Sacbrood virus (SBV) infects larvae of the honeybee (Apis mellifera), resulting in failure to pupate and death. Until now, identification of viruses in honeybee infections has been based on traditional methods such as electron microscopy, immunodiffusion, and enzyme-linked immunosorbent assay. Culture cannot be used because no honeybee cell lines are available. These techniques are low in sensitivity and specificity. However, the complete nucleotide sequence of SBV has recently been determined, and with these data, we now report a reverse transcription-PCR (RT-PCR) test for the direct, rapid, and sensitive detection of these viruses. RT-PCR was used to target five different areas of the SBV genome using infected honeybees and larvae originating from geographically distinct regions. The RT-PCR assay proved to be a rapid, specific, and sensitive diagnostic tool for the direct detection of SBV nucleic acid in samples of infected honeybees and brood regardless of geographic origin. The amplification products were sequenced, and phylogenetic analysis suggested the existence of at least three distinct genotypes of SBV.