Lau Katherine A, Theis Torsten, Gray Joanna, Rawlinson William D
Royal College of Pathologists of Australasia Quality Assurance Programs in Biosecurity, St. Leonards, NSW, Australia.
Serology and Virology Division, South Eastern Area Laboratory Services Microbiology, Prince of Wales Hospital, NSW Health Pathology, Sydney, NSW, Australia
J Clin Microbiol. 2017 Mar;55(3):783-790. doi: 10.1128/JCM.02173-16. Epub 2016 Dec 14.
The unprecedented 2015 Ebolavirus (EBOV) outbreak in West Africa was declared a public health emergency, making diagnosis and quality of testing a global issue. The accuracy of laboratory diagnostic capacity for EBOV was assessed in 2014 to 2016 using a proficiency testing (PT) strategy developed by the Royal College of Pathologists of Australasia Quality Assurance Programs (RCPAQAP) in Biosecurity. Following a literature search, EBOV-specific gene targets were ranked according to the frequency of their use in published methods. The most commonly used gene regions (nucleoprotein [NP], glycoprotein [GP], and RNA-dependent RNA polymerase [L]) were selected for the design of RNA transcripts to be included in the simulated EBOV specimens used for EBOV detection with PCR-based assays. Specimens were tested for stability and found to be stable on long-term storage (1 year) at -80°C and on shorter-term storage in lyophilized form (1 week at ambient temperature and a subsequent week at -80°C). These specimens were used in three EBOV PTs offered from April 2014 to March 2016. In the first and third PTs, all laboratories (3/3 and 9/9, respectively) correctly identified specimens containing EBOV RNA transcripts, while in the second PT, all but one laboratory (5/6) correctly confirmed the presence of EBOV. The EBOV PT panel was useful for ensuring the competency of laboratories in detecting EBOV in the absence of readily available clinical samples. The simulated EBOV specimen was safe, stable, and reliable and can be used in lyophilized form for future EBOV PT programs, allowing simplicity of transport.
2015年在西非爆发的前所未有的埃博拉病毒（EBOV）疫情被宣布为突发公共卫生事件，这使得诊断和检测质量成为一个全球性问题。2014年至2016年期间，使用澳大利亚皇家病理学家学院生物安全质量保证项目（RCPAQAP）制定的能力验证（PT）策略，对EBOV实验室诊断能力的准确性进行了评估。在文献检索之后，根据EBOV特异性基因靶点在已发表方法中的使用频率进行了排名。选择了最常用的基因区域（核蛋白[NP]、糖蛋白[GP]和RNA依赖性RNA聚合酶[L]）来设计RNA转录本，将其纳入用于基于PCR检测的模拟EBOV标本中。对标本进行了稳定性测试，发现其在-80°C下长期保存（1年）以及在冻干形式下短期保存（室温1周，随后在-80°C下保存1周）时均稳定。这些标本被用于2014年4月至2016年3月期间提供的三次EBOV PT测试。在第一次和第三次PT测试中,所有实验室（分别为3/3和9/9）均正确鉴定出含有EBOV RNA转录本的标本，而在第二次PT测试中，除一个实验室外（5/6），所有实验室均正确确认了EBOV的存在。EBOV PT测试组有助于确保实验室在缺乏现成临床样本的情况下检测EBOV的能力。模拟EBOV标本安全、稳定且可靠，可冻干形式用于未来的EBOV PT测试项目，便于运输。
