Luo Xue-Lian, Zhang Xiu-Dan, Li Bei-Jie, Qin Tian, Cao Zhi-Jie, Fan Qian-Jin, Yang Jing, Jin Dong, Lu Shan, Zheng Ya-Yun, Xu Xue-Fang, Pu Ji, Xu Jianguo
State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Changping, Beijing, China.
Department of Epidemiology, School of Public Health, Shanxi Medical University, Taiyuan, Shanxi Province, China.
Microbiol Spectr. 2023 Mar 28;11(2):e0501122. doi: 10.1128/spectrum.05011-22.
Lassa virus (LASV) is a causative agent of hemorrhagic fever epidemic in West Africa. In recent years, it has been transmitted several times to North America, Europe, and Asia. Standard reverse transcription (RT)-PCR and real-time RT-PCR are extensively used for early detection of LASV. However, the high nucleotide diversity of LASV strains complicates the development of appropriate diagnostic assays. Here, we analyzed LASV diversity clustered with geographic location and evaluated the specificity and sensitivity of two standard RT-PCR methods (GPC RT-PCR/1994 and 2007) and four commercial real-time RT-PCR kits (namely, Da an, Mabsky, Bioperfectus, and ZJ) to detect six representative LASV lineages using synthesized RNA templates. The results showed that the GPC RT-PCR/2007 assay had better sensitivity compared to the GPC RT-PCR/1994 assay. The Mabsky and ZJ kits were able to detect all RNA templates of six LASV lineages. Contrastingly, the Bioperfectus and Da an kits failed to detect lineages IV and V/VI. The limit of detection for lineage I with the Da an, Bioperfectus, and ZJ kits were significantly higher than that of the Mabsky kit at an RNA concentration of 1 × 10 to 1 × 10 copies/mL. The Bioperfectus and Da an kits detected lineages II and III at an RNA concentration of 1 × 10 copies/mL, higher than that of the other kits. In conclusion, the GPC RT-PCR/2007 assay and the Mabsky kit were suitable assays for the detection of LASV strains based on good analytical sensitivity and specificity. Lassa virus (LASV) is a significant human pathogen causing hemorrhagic fever in West Africa. Increased traveling around the world raises the risk of imported cases to other countries. The high nucleotide diversity of LASV strains clustered with geographic location complicates the development of appropriate diagnostic assays. In this study, we showed that the GPC reverse transcription (RT)-PCR/2007 assay and the Mabsky kit are suitable for detecting most LASV strains. Future assays for molecular detection of LASV should be based on specific countries/regions along with new variants.
拉沙病毒（LASV）是西非出血热疫情的病原体。近年来，它已多次传播至北美、欧洲和亚洲。标准逆转录（RT）-PCR和实时RT-PCR被广泛用于拉沙病毒的早期检测。然而，拉沙病毒毒株的高核苷酸多样性使得开发合适的诊断检测方法变得复杂。在此，我们分析了与地理位置相关的拉沙病毒多样性，并使用合成RNA模板评估了两种标准RT-PCR方法（GPC RT-PCR/1994和2007）以及四种商业实时RT-PCR试剂盒（即达安、Mabsky、BioPerfectus和ZJ）检测六种代表性拉沙病毒谱系的特异性和灵敏度。结果表明，与GPC RT-PCR/1994检测方法相比，GPC RT-PCR/2007检测方法具有更高的灵敏度。Mabsky和ZJ试剂盒能够检测六种拉沙病毒谱系的所有RNA模板。相比之下，BioPerfectus和达安试剂盒未能检测到谱系IV和V/VI。在RNA浓度为1×10至1×10拷贝/毫升时，达安、BioPerfectus和ZJ试剂盒对谱系I的检测限显著高于Mabsky试剂盒。BioPerfectus和达安试剂盒在RNA浓度为1×10拷贝/毫升时检测到谱系II和III，高于其他试剂盒。总之，基于良好的分析灵敏度和特异性，GPC RT-PCR/2007检测方法和Mabsky试剂盒是检测拉沙病毒毒株的合适检测方法。拉沙病毒（LASV）是导致西非出血热的重要人类病原体。全球旅行增加使其他国家出现输入性病例的风险上升。与地理位置相关的拉沙病毒毒株的高核苷酸多样性使得开发合适的诊断检测方法变得复杂。在本研究中，我们表明GPC逆转录（RT）-PCR/2007检测方法和Mabsky试剂盒适用于检测大多数拉沙病毒毒株。未来用于拉沙病毒分子检测的检测方法应基于特定国家/地区以及新的病毒变种。
