Ayhan Nazli, Baronti Cecile, Thrion Laurence, Bongiorno Gioia, Maia Carla, Charrel Remi N
Unite des Virus Emergents (UVE), L Université d Aix-Marseille-Universita di Corsica, IRD 190, Inserm 1207-L Institut de recherche biomédicale des armées (IRBA), 19-21, Bd. Jean Moulin, 13005, Marseille, France.
Centre National de Reference des Arbovirus, Inserm-IRBA, Marseille, France.
Parasit Vectors. 2025 May 12;18(1):173. doi: 10.1186/s13071-025-06785-0.
Sand fly-borne phleboviruses (SbPV) are globally distributed and pose potential public health risks. Despite increased detection in recent decades, detailed knowledge of their ecology, characteristics and clinical relevance remains limited. Many cases of SbPV infection likely go unreported or misdiagnosed due to limited awareness and the lack of standardized screening. The External Quality Assessment (EQA) reported herein was organized within the framework of the European Union CLIMOS (EU Climate Monitoring and Decision Support Framework for Sand Fly-borne Diseases Detection and Mitigation) project. The aim of this EQA was to standardize the detection of phleboviruses in order to provide comparable data to feed mathematical models for the surveillance of the impact of climate changes and environmental parameters on the kinetics and diversity of sand fly species and on sand fly-borne microorganisms.
Nine laboratories from seven countries participated in the EQA. Each laboratory was provided with eight vials, each containing an anonymous sample; two vials of lyophilized primers and probes to be used for the detection of Toscana virus (TOSV) and several Sandfly fever Sicilian virus (SFSV) species with a reverse-transcriptase PCR (RT-PCR) assay; and one vial of lyophilized primers for the detection of generic phleboviruses with a RT-PCR assay along with the standard operating procedure. The laboratories were instructed to submit their results together with details on the techniques employed.
All nine laboratories successfully detected the two TOSV- and the one SFSV-positive samples. Only one laboratory, using a generic phlebovirus assay, detected all of the targeted phleboviruses.
All participating laboratories successfully identified the two TOSV and one SFSV using the proposed RT-qPCR assays, albeit with some variations in cycle threshold values across laboratories. The detection rate of SbPV was lower with the generic Phlebovirus assay than with the specific real-time RT-qPCR assays. This EQA aimed to assess the SbPV detection capabilities of molecular tools and strengthen their use, thereby supporting the involvement of laboratories in virus discovery and surveillance beyond their core expertise.
白蛉传播的静脉病毒（SbPV）在全球范围内分布，对公众健康构成潜在风险。尽管近几十年来检测有所增加，但对其生态学、特征和临床相关性的详细了解仍然有限。由于认识不足和缺乏标准化筛查，许多SbPV感染病例可能未被报告或误诊。本文报告的外部质量评估（EQA）是在欧盟CLIMOS（欧盟白蛉传播疾病检测与缓解气候监测和决策支持框架）项目框架内组织的。该EQA的目的是规范静脉病毒的检测，以便提供可比数据，为数学模型提供信息，用于监测气候变化和环境参数对白蛉物种动力学和多样性以及对白蛉传播微生物的影响。
来自七个国家的九个实验室参加了EQA。每个实验室收到八个小瓶，每个小瓶含有一个匿名样本；两个冻干引物和探针小瓶，用于通过逆转录酶聚合酶链反应（RT-PCR）检测托斯卡纳病毒（TOSV）和几种西西里白蛉热病毒（SFSV）；以及一个冻干引物小瓶，用于通过RT-PCR检测通用静脉病毒以及标准操作程序。要求各实验室提交结果以及所采用技术的详细信息。
所有九个实验室均成功检测出两个TOSV阳性样本和一个SFSV阳性样本。只有一个实验室使用通用静脉病毒检测法检测出了所有目标静脉病毒。
所有参与实验室均使用提议的RT-qPCR检测法成功鉴定出两种TOSV和一种SFSV，尽管各实验室的循环阈值存在一些差异。通用静脉病毒检测法检测SbPV的检出率低于特异性实时RT-qPCR检测法。该EQA旨在评估分子工具检测SbPV的能力并加强其应用，从而支持实验室参与超出其核心专业领域的病毒发现和监测工作。
