Bonney Laura C, Watson Robert J, Afrough Babak, Mullojonova Manija, Dzhuraeva Viktoriya, Tishkova Farida, Hewson Roger
National Infection Service, Public Health England, Porton Down, Salisbury, United Kingdom.
Department of Virology, Tajik Research Institute of Preventive Medicine of the Ministry of Health of the Republic of Tajikistan, Dushanbe, Republic of Tajikistan.
PLoS Negl Trop Dis. 2017 Oct 13;11(10):e0006013. doi: 10.1371/journal.pntd.0006013. eCollection 2017 Oct.
Crimean-Congo Haemorrhagic fever Virus (CCHFV) is a rapidly emerging vector-borne pathogen and the cause of a virulent haemorrhagic fever affecting large parts of Europe, Africa, the Middle East and Asia.
METHODOLOGY/PRINCIPLE FINDINGS: An isothermal recombinase polymerase amplification (RPA) assay was successfully developed for molecular detection of CCHFV. The assay showed rapid (under 10 minutes) detection of viral extracts/synthetic virus RNA of all 7 S-segment clades of CCHFV, with high target specificity. The assay was shown to tolerate the presence of inhibitors in crude preparations of mock field samples, indicating that this assay may be suitable for use in the field with minimal sample preparation. The CCHFV RPA was successfully used to screen and detect CCHFV positives from a panel of clinical samples from Tajikistan.
CONCLUSIONS/SIGNIFICANCE: The assay is a rapid, isothermal, simple-to-perform molecular diagnostic, which can be performed on a light, portable real-time detection device. It is ideally placed therefore for use as a field-diagnostic or in-low resource laboratories, for monitoring of CCHF outbreaks at the point-of-need, such as in remote rural regions in affected countries.
克里米亚-刚果出血热病毒（CCHFV）是一种迅速出现的媒介传播病原体，可导致一种烈性出血热，影响欧洲、非洲、中东和亚洲的大片地区。
方法/主要发现：成功开发了一种等温重组酶聚合酶扩增（RPA）检测方法，用于CCHFV的分子检测。该检测方法能快速（10分钟内）检测CCHFV所有7个S片段分支的病毒提取物/合成病毒RNA，具有高靶标特异性。该检测方法可耐受模拟野外样本粗提物中抑制剂的存在，表明该检测方法在样本制备最少的情况下可能适用于野外检测。CCHFV RPA检测方法成功用于筛选和检测来自塔吉克斯坦的一组临床样本中的CCHFV阳性样本。
结论/意义：该检测方法是一种快速、等温、操作简单的分子诊断方法，可在轻便、便携式实时检测设备上进行。因此，它非常适合用作现场诊断或在资源匮乏的实验室中，在需要时（如在受影响国家的偏远农村地区）监测CCHF疫情。
