TY - JOUR
T1 - Development of a multiplex RT-PCR for the simultaneous detection of three viruses of the honeybee (Apis mellifera L.)
T2 - Acute bee paralysis virus, Black queen cell virus and Sacbrood virus
AU - Grabensteiner, Elvira
AU - Bakonyi, Tamás
AU - Ritter, Wolfgang
AU - Pechhacker, Hermann
AU - Nowotny, Norbert
N1 - Funding Information:
This work was funded by a grant of the Austrian Federal Office and Research Centre for Agriculture (BFL), No. 921/99, and grants of the Hungarian Scientific Research Fund (OTKA) Nos. F043155 and D048647. The authors wish to thank Dr. Irmgard Derakhshifar (Institute for Apiculture, Austrian Federal Office and Research Centre for Agriculture, Vienna) for providing the pre-tested Austrian ABPV, BQCV and SBV samples used in this study.
PY - 2007/3
Y1 - 2007/3
N2 - A single-step multiple-target (multiplex) reverse transcription-PCR (RT-PCR) was developed for the simultaneous detection and differentiation of three economically important viruses of the honeybee Apis mellifera L.: Acute bee paralysis virus (ABPV), Black queen cell virus (BQCV) and Sacbrood virus (SBV). Three compatible sets of primers, specific for each virus, were designed in conserved regions of the viral genomes for use in a one-step (single tube) RT-PCR assay. The individual RT-PCR assays and the combined multiplex assay were optimized for highest sensitivity and specificity. The multiplex RT-PCR assay was tested on field samples collected from Austrian honeybee colonies. All three viruses were detected, and their identity was confirmed by sequencing of the PCR products. The described multiplex RT-PCR proved to be an accurate tool for rapid simultaneous detection of ABPV, BQCV and SBV directly in honeybee specimens.
AB - A single-step multiple-target (multiplex) reverse transcription-PCR (RT-PCR) was developed for the simultaneous detection and differentiation of three economically important viruses of the honeybee Apis mellifera L.: Acute bee paralysis virus (ABPV), Black queen cell virus (BQCV) and Sacbrood virus (SBV). Three compatible sets of primers, specific for each virus, were designed in conserved regions of the viral genomes for use in a one-step (single tube) RT-PCR assay. The individual RT-PCR assays and the combined multiplex assay were optimized for highest sensitivity and specificity. The multiplex RT-PCR assay was tested on field samples collected from Austrian honeybee colonies. All three viruses were detected, and their identity was confirmed by sequencing of the PCR products. The described multiplex RT-PCR proved to be an accurate tool for rapid simultaneous detection of ABPV, BQCV and SBV directly in honeybee specimens.
KW - Acute bee paralysis virus
KW - Apis mellifera
KW - Black queen cell virus
KW - Honeybee
KW - Multiplex RT-PCR
KW - Sacbrood virus
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U2 - 10.1016/j.jip.2006.11.006
DO - 10.1016/j.jip.2006.11.006
M3 - Article
C2 - 17207498
AN - SCOPUS:33846965959
SN - 0022-2011
VL - 94
SP - 222
EP - 225
JO - Journal of Invertebrate Pathology
JF - Journal of Invertebrate Pathology
IS - 3
ER -