Abstract：
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 in vitro 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 × 1010 to 1 × 1011 copies/mL. The Bioperfectus and Da an kits detected lineages II and III at an RNA concentration of 1 × 109 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. IMPORTANCE 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.
摘要：
拉沙病毒（LASV）是西非出血热流行的病原体。近年来,它已经多次传播到北美，欧洲,和亚洲。标准逆转录(RT)-PCR和实时RT-PCR广泛用于LASV的早期检测。然而,LASV毒株的高核苷酸多样性使适当的诊断试验的发展变得复杂.这里,我们分析了具有地理位置的LASV多样性，并评估了两种标准RT-PCR方法（GPCRT-PCR/1994和2007）和四种商业实时RT-PCR试剂盒（即，大安,马布斯基,Bioperfectus,和ZJ）使用体外合成的RNA模板检测六个代表性的LASV谱系。结果表明,与GPCRT-PCR/1994测定相比,GPCRT-PCR/2007测定具有更好的灵敏度。Mabsky和ZJ试剂盒能够检测六个LASV谱系的所有RNA模板。相反，Bioperfectus和Daan试剂盒未能检测到谱系IV和V/VI。Daan对谱系I的检测限，Bioperfectus,和ZJ试剂盒在RNA浓度为1×1010至1×1011拷贝/mL时显著高于Mabsky试剂盒。Bioperfectus和Daan试剂盒以1×109拷贝/mL的RNA浓度检测到谱系II和III，高于其他套件。总之,基于良好的分析灵敏度和特异性,GPCRT-PCR/2007检测和Mabsky试剂盒是检测LASV毒株的合适检测方法.重要性拉沙病毒(LASV)是引起西非出血热的重要人类病原体。世界各地旅行的增加增加了向其他国家输入病例的风险。聚集在地理位置上的LASV菌株的高核苷酸多样性使适当的诊断测定的开发变得复杂。在这项研究中,我们表明GPC逆转录（RT）-PCR/2007检测和Mabsky试剂盒适用于检测大多数LASV菌株。LASV分子检测的未来测定应基于特定国家/地区以及新的变体。
