Development of a multiplex RT-PCR for the simultaneous detection of three viruses of the honeybee (Apis mellifera L.)

Acute bee paralysis virus, Black queen cell virus and Sacbrood virus

Elvira Grabensteiner, Tamás Bakonyi, Wolfgang Ritter, Hermann Pechhacker, Norbert Nowotny

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)222-225
Number of pages4
JournalJournal of Invertebrate Pathology
Volume94
Issue number3
DOIs
Publication statusPublished - Mar 2007

Fingerprint

Sacbrood virus
Acute bee paralysis virus
Black queen cell virus
honeybee
Apis mellifera
bee
honey bees
virus
reverse transcriptase polymerase chain reaction
viruses
assays
assay
honey bee colonies
detection
genome

Keywords

  • Acute bee paralysis virus
  • Apis mellifera
  • Black queen cell virus
  • Honeybee
  • Multiplex RT-PCR
  • Sacbrood virus

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Insect Science

Cite this

Development of a multiplex RT-PCR for the simultaneous detection of three viruses of the honeybee (Apis mellifera L.) : Acute bee paralysis virus, Black queen cell virus and Sacbrood virus. / Grabensteiner, Elvira; Bakonyi, Tamás; Ritter, Wolfgang; Pechhacker, Hermann; Nowotny, Norbert.

In: Journal of Invertebrate Pathology, Vol. 94, No. 3, 03.2007, p. 222-225.

Research output: Contribution to journalArticle

@article{00bfc779527746a5b51d708ff8a2bea7,
title = "Development of a multiplex RT-PCR for the simultaneous detection of three viruses of the honeybee (Apis mellifera L.): Acute bee paralysis virus, Black queen cell virus and Sacbrood virus",
abstract = "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.",
keywords = "Acute bee paralysis virus, Apis mellifera, Black queen cell virus, Honeybee, Multiplex RT-PCR, Sacbrood virus",
author = "Elvira Grabensteiner and Tam{\'a}s Bakonyi and Wolfgang Ritter and Hermann Pechhacker and Norbert Nowotny",
year = "2007",
month = "3",
doi = "10.1016/j.jip.2006.11.006",
language = "English",
volume = "94",
pages = "222--225",
journal = "Journal of Invertebrate Pathology",
issn = "0022-2011",
publisher = "Academic Press Inc.",
number = "3",

}

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

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

UR - http://www.scopus.com/inward/record.url?scp=33846965959&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33846965959&partnerID=8YFLogxK

U2 - 10.1016/j.jip.2006.11.006

DO - 10.1016/j.jip.2006.11.006

M3 - Article

VL - 94

SP - 222

EP - 225

JO - Journal of Invertebrate Pathology

JF - Journal of Invertebrate Pathology

SN - 0022-2011

IS - 3

ER -