Victorian Infectious Diseases Reference Laboratory, Royal Melbourne Hospital, at the Peter Doherty Institute for Infection and Immunity, Vic 3000, Australia.
Victorian Infectious Diseases Reference Laboratory, Royal Melbourne Hospital, at the Peter Doherty Institute for Infection and Immunity, Vic 3000, Australia; Department of Infectious Diseases, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Vic 3000, Australia.
J Clin Virol. 2023 Apr;161:105424. doi: 10.1016/j.jcv.2023.105424. Epub 2023 Mar 17.
The current global mpox virus (MPXV) outbreak has been declared a Public Health Emergency of International Concern by WHO, with more than 80,000 cases confirmed across multiple continents. Diagnosis is confirmed by PCR of viral DNA from vesicle and other swabs.
The aim of this study was to assess commercial RT PCR assays for Orthopoxvirus (OPX) and MPXV for analytical sensitivity, and percent agreements and compare them to primer/probe sets employed at the Victorian Infectious Diseases Reference Laboratory (VIDRL), Centers for Disease Control andPrevention (CDC) and US Army Medical Research Institute of Infectious Diseases (USAMRIID). Limits of detection (LOD), intra-run variability, cross-reactivity and performance on forty clinical samples was assessed on eleven commercial assays and five primer/probe combinations used at VIDRL, CDC and USAMRIID.
All assays were able to detect OPX and MPXV (LOD 57 to 14,495 copies/mL) with intra-run coefficients of variation between Cycle thresholds of 0.58 and 3.44, and there was no unexpected cross-reactivity. All assays demonstrated 100% negative percent agreement with clinical samples and all but one yielded 100% positive percent agreement.
Variations in LOD between assays may be dependent on the platform used and sample type. Despite the overall comparable performance of the assays assessed, it is important that routine laboratories perform in-house validations before implementing RT PCR for OPX and/or MPXV as reliable and accurate laboratory diagnosis of MPXV and isolation is crucial to containing the spread of this current outbreak and informing public health interventions and response.
目前，世界卫生组织已宣布全球猴痘病毒（MPXV）疫情为国际关注的突发公共卫生事件，多个大洲已确认超过 8 万例病例。诊断通过从水疱和其他拭子中提取病毒 DNA 进行 PCR 来确认。
本研究旨在评估商用 RT-PCR 检测试剂盒对正痘病毒（OPX）和 MPXV 的分析灵敏度，并评估其与维多利亚传染病参考实验室（VIDRL）、美国疾病控制与预防中心（CDC）和美国陆军传染病医学研究所（USAMRIID）使用的引物/探针的符合率，并进行比较。在 11 种商业检测试剂盒和 VIDRL、CDC 和 USAMRIID 使用的 5 种引物/探针组合上，评估了 40 份临床样本的检测限（LOD）、批内变异性、交叉反应性和性能。
所有检测试剂盒均能检测到 OPX 和 MPXV（LOD 57 至 14495 拷贝/ml），其循环阈值的批内变异系数在 0.58 至 3.44 之间，且无意外的交叉反应性。所有检测试剂盒与临床样本的阴性符合率均为 100%，除 1 种外，其余检测试剂盒的阳性符合率均为 100%。
检测试剂盒之间 LOD 的差异可能取决于所使用的平台和样本类型。尽管评估的检测试剂盒整体性能相当，但常规实验室在实施 OPX 和/或 MPXV 的 RT-PCR 之前进行内部验证非常重要，因为可靠且准确的 MPXV 实验室诊断和分离对于控制当前疫情的传播以及为公共卫生干预和应对提供信息至关重要。
