Robardet Emmanuelle, Demerson Jean-Michel, Andrieu Sabrina, Cliquet Florence
French Agency for Food, Environmental and Occupational Health & Safety (ANSES), Nancy Laboratory for Rabies and Wildlife, World Health Organization Collaborating Centre for Research and Management in Zoonoses Control, Technopôle Agricole et Vétérinaire, BP 40009, 54220 Malzéville cedex, France.
J Wildl Dis. 2012 Oct;48(4):858-68. doi: 10.7589/2011-07-205.
The first European interlaboratory comparison of tetracycline and age determination with red fox (Vulpes vulpes) tooth samples was organized by the European Union Reference Laboratory for rabies. Performance and procedures implemented by member states were compared. These techniques are widely used to monitor bait uptake in European oral rabies vaccination campaigns. A panel of five red fox half-mandibles comprising one weak positive juvenile sample, two positive adult samples, one negative juvenile sample, and one negative adult sample were sent, along with a technical questionnaire, to 12 laboratories participating on a voluntary basis. The results of only three laboratories (25%) were 100% correct. False-negative results were more frequently seen in weak positive juvenile samples (58%) but were infrequent in positive adult samples (4%), probably due to differences in the ease of reading the two groups of teeth. Four laboratories (44%) had correct results for age determination on all samples. Ages were incorrectly identified in both adult and juvenile samples, with 11 and 17% of discordant results, respectively. Analysis of the technical questionnaires in parallel with test results suggested that all laboratories cutting mandible sections between the canine and first premolar obtained false results. All the laboratories using longitudinal rather than transverse sections and those not using a mounting medium also produced false results. Section thickness appeared to affect the results; no mistakes were found in laboratories using sections <150 μm thick. Factors having a potential impact on the success of laboratories were discussed, and recommendations proposed. Such interlaboratory trials underline the importance of using standardized procedures for biomarker detection in oral rabies vaccination campaigns. Several changes can be made to improve analysis quality and increase the comparability of bait uptake frequencies among member states.
欧洲狂犬病参考实验室组织了首次针对赤狐（Vulpes vulpes）牙齿样本进行四环素和年龄测定的欧洲实验室间比对。对成员国实施的操作性能和程序进行了比较。这些技术被广泛用于监测欧洲口服狂犬病疫苗接种运动中的诱饵摄取情况。一组包含一个弱阳性幼年样本、两个阳性成年样本、一个阴性幼年样本和一个阴性成年样本的五只赤狐半下颌骨，连同一份技术问卷，被发送给了12个自愿参与的实验室。只有三个实验室（25%）的结果100%正确。在弱阳性幼年样本中更常出现假阴性结果（58%），但在阳性成年样本中较少见（4%），这可能是由于两组牙齿在读取难度上存在差异。四个实验室（44%）对所有样本的年龄测定结果正确。成年和幼年样本的年龄都被错误识别，不一致结果分别为11%和17%。将技术问卷分析与测试结果并行分析表明，所有在犬齿和第一前磨牙之间切割下颌骨切片的实验室都得到了错误结果。所有使用纵向而非横向切片以及未使用固定介质的实验室也都产生了错误结果。切片厚度似乎会影响结果；在使用厚度<150μm切片的实验室中未发现错误。讨论了对实验室成功有潜在影响的因素，并提出了建议。此类实验室间试验强调了在口服狂犬病疫苗接种运动中使用标准化程序进行生物标志物检测的重要性。可以做出一些改变以提高分析质量并增加成员国之间诱饵摄取频率的可比性。
