Detection of acute bee paralysis virus and black queen cell virus from honeybees by reverse transcriptase pcr.
A reverse transcriptase PCR (RT-PCR) assay was developed for the detection of acute bee paralysis virus (ABPV) and black queen cell virus (BQCV), two honeybee viruses. Complete genome sequences were used to design unique PCR primers within a 1-kb region from the 3' end of both genomes to amplify a fragment of 900 bp from ABPV and 700 bp from BQCV. The combined guanidinium thiocyanate and silica membrane method was used to extract total RNA from samples of healthy and laboratory-infected bee pupae. In a blind test, RT-PCR successfully identified the samples containing ABPV and BQCV. Sensitivities were approximately 1,600 genome equivalents of purified ABPV and 130 genome equivalents of BQCV.
Benjeddou
M
M
Department of Microbiology, University of the Western Cape, Bellville 7535, Cape Town, South Africa.
Leat
N
N
Allsopp
M
M
Davison
S
S
D000818:Animals; D001516:Bees / Q000254:growth & development / Q000821:virology*; D007304:Insect Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010849:Picornaviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D012367:RNA, Viral