Intra-laboratory validation of chronic bee paralysis virus quantitation using an accredited standardised real-time quantitative RT-PCR method.
Chronic bee paralysis virus (CBPV) is responsible for chronic bee paralysis, an infectious and contagious disease in adult honey bees (Apis mellifera L.). A real-time RT-PCR assay to quantitate the CBPV load is now available. To propose this assay as a reference method, it was characterised further in an intra-laboratory study during which the reliability and the repeatability of results and the performance of the assay were confirmed. The qPCR assay alone and the whole quantitation method (from sample RNA extraction to analysis) were both assessed following the ISO/IEC 17025 standard and the recent XP U47-600 standard issued by the French Standards Institute. The performance of the qPCR assay and of the overall CBPV quantitation method were validated over a 6 log range from 10(2) to 10(8) with a detection limit of 50 and 100 CBPV RNA copies, respectively, and the protocol of the real-time RT-qPCR assay for CBPV quantitation was approved by the French Accreditation Committee.
French Agency for Food, Environmental and Occupational Health & Safety, Sophia-Antipolis Laboratory, Bee Diseases Unit, BP 111, 06902 Sophia Antipolis, France. philippe.blanchard@anses.fr
Philippe
P
Blanchard
Julie
J
Regnault
Frank
F
Schurr
Eric
E
Dubois
Magali
M
Ribière
D000068:Accreditation; D000818:Animals; D001516:Bees / Q000821:virology*; D002908:Chronic Disease; D007304:Insect Viruses / Q000235:genetics* / Q000302:isolation & purification; D007753:Laboratories / Q000592:standards*; D010243:Paralysis / Q000821:virology; D012328:RNA Viruses / Q000235:genetics* / Q000302:isolation & purification; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D012680:Sensitivity and Specificity
D012367:RNA, Viral