
Trueness and precision of the real-time RT-PCR method for quantifying the chronic bee paralysis virus genome in bee homogenates evaluated by a comparative inter-laboratory study
- Author
- Frank Schurr, Nicolas Cougoule, Marie-Pierre Riviere, Magali Ribiere-Chabert, Hamid Achour, Dan Adam, Carlos Castillo, Dirk de Graaf (UGent) , Eva Forsgren, Anna Granato, Sirpa Heinikainen, Julia Jurovcikova, Per Kryger, Christine Manson, Marie-Francoise Menard, Stephane Perennes, Marc O Schaefer, Elena San Miguel Ibanez, Joao Silva, Ivana Tlak Gajger, Victoria Tomkies, Ivan Toplak, Alain Viry, Dagmara Zdanska and Eric Dubois
- Organization
- Abstract
- The Chronic bee paralysis virus (CBPV) is the aetiological agent of chronic bee paralysis, a contagious disease associated with nervous disorders in adult honeybees leading to massive mortalities in front of the hives. Some of the clinical signs frequently reported, such as trembling, may be confused with intoxication syndromes. Therefore, laboratory diagnosis using real-time PCR to quantify CBPV loads is used to confirm disease. Clinical signs of chronic paralysis are usually associated with viral loads higher than 108 copies of CBPV genome copies per bee (8 log(10) CBPV/bee). This threshold is used by the European Union Reference Laboratory for Bee Health to diagnose the disease. In 2015, the accuracy of measurements of three CBPV loads (5, 8 and 9 log(10) CBPV/bee) was assessed through an inter-laboratory study. Twenty-one participants, including 16 European National Reference Laboratories, received 13 homogenates of CBPV-infected bees adjusted to the three loads. Participants were requested to use the method usually employed for routine diagnosis. The quantitative results (n = 270) were analysed according to international standards NF ISO 13528 (2015) and NF ISO 5725-2 (1994). The standard deviations of measurement reproducibility (S-R) were 0.83, 1.06 and 1.16 at viral loads 5, 8 and 9 log(10) CBPV/bee, respectively. The inter-laboratory confidence of viral quantification (+/- 1.96 S-R) at the diagnostic threshold (8 log(10) CBPV/bee) was +/- 2.08 log(10) CBPV/bee. These results highlight the need to take into account the confidence of measurements in epidemiological studies using results from different laboratories. Considering this confidence, viral loads over 6 log(10) CBPV/bee may be considered to indicate probable cases of chronic paralysis.
- Keywords
- quantitative PCR, honeybee, RNA virus, ring test, accuracy, APIS-MELLIFERA L., QUANTITATION, INFECTION
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Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-8636903
- MLA
- Schurr, Frank, et al. “Trueness and Precision of the Real-Time RT-PCR Method for Quantifying the Chronic Bee Paralysis Virus Genome in Bee Homogenates Evaluated by a Comparative Inter-Laboratory Study.” JOURNAL OF VIROLOGICAL METHODS, vol. 248, 2017, pp. 217–25, doi:10.1016/j.jviromet.2017.07.013.
- APA
- Schurr, F., Cougoule, N., Riviere, M.-P., Ribiere-Chabert, M., Achour, H., Adam, D., … Dubois, E. (2017). Trueness and precision of the real-time RT-PCR method for quantifying the chronic bee paralysis virus genome in bee homogenates evaluated by a comparative inter-laboratory study. JOURNAL OF VIROLOGICAL METHODS, 248, 217–225. https://doi.org/10.1016/j.jviromet.2017.07.013
- Chicago author-date
- Schurr, Frank, Nicolas Cougoule, Marie-Pierre Riviere, Magali Ribiere-Chabert, Hamid Achour, Dan Adam, Carlos Castillo, et al. 2017. “Trueness and Precision of the Real-Time RT-PCR Method for Quantifying the Chronic Bee Paralysis Virus Genome in Bee Homogenates Evaluated by a Comparative Inter-Laboratory Study.” JOURNAL OF VIROLOGICAL METHODS 248: 217–25. https://doi.org/10.1016/j.jviromet.2017.07.013.
- Chicago author-date (all authors)
- Schurr, Frank, Nicolas Cougoule, Marie-Pierre Riviere, Magali Ribiere-Chabert, Hamid Achour, Dan Adam, Carlos Castillo, Dirk de Graaf, Eva Forsgren, Anna Granato, Sirpa Heinikainen, Julia Jurovcikova, Per Kryger, Christine Manson, Marie-Francoise Menard, Stephane Perennes, Marc O Schaefer, Elena San Miguel Ibanez, Joao Silva, Ivana Tlak Gajger, Victoria Tomkies, Ivan Toplak, Alain Viry, Dagmara Zdanska, and Eric Dubois. 2017. “Trueness and Precision of the Real-Time RT-PCR Method for Quantifying the Chronic Bee Paralysis Virus Genome in Bee Homogenates Evaluated by a Comparative Inter-Laboratory Study.” JOURNAL OF VIROLOGICAL METHODS 248: 217–225. doi:10.1016/j.jviromet.2017.07.013.
- Vancouver
- 1.Schurr F, Cougoule N, Riviere M-P, Ribiere-Chabert M, Achour H, Adam D, et al. Trueness and precision of the real-time RT-PCR method for quantifying the chronic bee paralysis virus genome in bee homogenates evaluated by a comparative inter-laboratory study. JOURNAL OF VIROLOGICAL METHODS. 2017;248:217–25.
- IEEE
- [1]F. Schurr et al., “Trueness and precision of the real-time RT-PCR method for quantifying the chronic bee paralysis virus genome in bee homogenates evaluated by a comparative inter-laboratory study,” JOURNAL OF VIROLOGICAL METHODS, vol. 248, pp. 217–225, 2017.
@article{8636903, abstract = {{The Chronic bee paralysis virus (CBPV) is the aetiological agent of chronic bee paralysis, a contagious disease associated with nervous disorders in adult honeybees leading to massive mortalities in front of the hives. Some of the clinical signs frequently reported, such as trembling, may be confused with intoxication syndromes. Therefore, laboratory diagnosis using real-time PCR to quantify CBPV loads is used to confirm disease. Clinical signs of chronic paralysis are usually associated with viral loads higher than 108 copies of CBPV genome copies per bee (8 log(10) CBPV/bee). This threshold is used by the European Union Reference Laboratory for Bee Health to diagnose the disease. In 2015, the accuracy of measurements of three CBPV loads (5, 8 and 9 log(10) CBPV/bee) was assessed through an inter-laboratory study. Twenty-one participants, including 16 European National Reference Laboratories, received 13 homogenates of CBPV-infected bees adjusted to the three loads. Participants were requested to use the method usually employed for routine diagnosis. The quantitative results (n = 270) were analysed according to international standards NF ISO 13528 (2015) and NF ISO 5725-2 (1994). The standard deviations of measurement reproducibility (S-R) were 0.83, 1.06 and 1.16 at viral loads 5, 8 and 9 log(10) CBPV/bee, respectively. The inter-laboratory confidence of viral quantification (+/- 1.96 S-R) at the diagnostic threshold (8 log(10) CBPV/bee) was +/- 2.08 log(10) CBPV/bee. These results highlight the need to take into account the confidence of measurements in epidemiological studies using results from different laboratories. Considering this confidence, viral loads over 6 log(10) CBPV/bee may be considered to indicate probable cases of chronic paralysis.}}, author = {{Schurr, Frank and Cougoule, Nicolas and Riviere, Marie-Pierre and Ribiere-Chabert, Magali and Achour, Hamid and Adam, Dan and Castillo, Carlos and de Graaf, Dirk and Forsgren, Eva and Granato, Anna and Heinikainen, Sirpa and Jurovcikova, Julia and Kryger, Per and Manson, Christine and Menard, Marie-Francoise and Perennes, Stephane and Schaefer, Marc O and San Miguel Ibanez, Elena and Silva, Joao and Gajger, Ivana Tlak and Tomkies, Victoria and Toplak, Ivan and Viry, Alain and Zdanska, Dagmara and Dubois, Eric}}, issn = {{0166-0934}}, journal = {{JOURNAL OF VIROLOGICAL METHODS}}, keywords = {{quantitative PCR,honeybee,RNA virus,ring test,accuracy,APIS-MELLIFERA L.,QUANTITATION,INFECTION}}, language = {{eng}}, pages = {{217--225}}, title = {{Trueness and precision of the real-time RT-PCR method for quantifying the chronic bee paralysis virus genome in bee homogenates evaluated by a comparative inter-laboratory study}}, url = {{http://doi.org/10.1016/j.jviromet.2017.07.013}}, volume = {{248}}, year = {{2017}}, }
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