In the UK, research on hazard group 4 (HG4) pathogens requires specialised Containment Level 4 (CL4) facilities. These differ from Biosafety Level 4 (BSL4) conditions in that work is conducted in class III microbiological safety cabinets for primary containment instead of using positive pressure suits. This presents unique challenges associated with the physical restrictions of working in a limited space, and prohibits the use of many techniques and specialist equipment. In consequence, detailed studies on the biology of HG4 pathogens and in particular their immunological relationships with the host are understudied in the UK; for example, the majority of immunological assays with which the immune system is interrogated require specialist equipment that is unsuitable for CL4. Multiplexing to simultaneously measure multiple analytes is increasingly being used in immunological studies. This assay is attractive for CL4 work because it reduces the time spent in the laboratory whilst maximising the use of valuable sample volume. The Luminex microsphere approach allows for the determination of many cytokines and chemokines, however, the detection system uses fixed aligned lasers and integrated computer systems which are unsuitable for use at CL4. Therefore, we have developed an approach in which the Luminex assay is conducted within the CL4 laboratory and a formalin-fixation stage is introduced to allow for analysis to be undertaken outside of containment. Quality control preparations allow the assay characteristics to be monitored and analysis of assay performance to be evaluated. Our data demonstrate that Luminex is an applicable tool for use at CL4 and that assays can be run reliably to generate reproducible standardised data across different plates and individual experiments.

中文翻译：

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多重微珠检测 (Luminex) 在 4 级遏制级别的使用和可靠性

在英国，对危险组 4 (HG4) 病原体的研究需要专门的遏制级别 4 (CL4) 设施。这些不同于生物安全 4 级 (BSL4) 条件，因为工作是在 III 级微生物安全柜中进行的，而不是使用正压防护服。这带来了与在有限空间内工作的物理限制相关的独特挑战，并禁止使用许多技术和专业设备。因此，英国对 HG4 病原体的生物学，特别是它们与宿主的免疫关系的详细研究没有得到充分研究。例如，用于询问免疫系统的大多数免疫学分析需要不适合 CL4 的专业设备。同时测量多种分析物的多重检测越来越多地用于免疫学研究。这种检测对于 CL4 工作很有吸引力，因为它减少了在实验室中花费的时间，同时最大限度地利用了有价值的样品量。Luminex 微球方法允许测定许多细胞因子和趋化因子，但是，检测系统使用不适合用于 CL4 的固定对准激光器和集成计算机系统。因此，我们开发了一种方法，其中 Luminex 检测在 CL4 实验室内进行，并引入了福尔马林固定阶段，以便在安全壳外进行分析。质量控制准备允许监测化验特性并评估化验性能的分析。