RT-PCR assay for detection of Lassa virus and related Old World arenaviruses targeting the L gene
RT-PCR assay for detection of Lassa virus and related Old World arenaviruses targeting the L gene
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DOI：
10.1016/j.trstmh.2005.03.018
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发表时间：
2007-12-01
影响因子：
2.2
通讯作者：
Guenther, Stephan
中科院分区：
文献类型：
--
作者：
Vieth, Simon;Drosten, Christian;Guenther, Stephan
本研究描述了一种针对沙粒病毒L RNA片段的RT-PCR检测方法。根据已发表的拉沙病毒、淋巴细胞脉络丛脑膜炎病毒（LCMV）、皮丘德病毒和Tacaribe病毒的序列，以及本研究确定的拉沙病毒、LCMV、Ippy病毒、Mobala病毒和Mopeia病毒的15个新序列，确定了L基因聚合酶结构域的保守区域。以这些区域为靶点，开发并优化了检测所有已知旧大陆沙粒病毒的PCR检测方法。在一组重复试验（95%检测限）中产生95%阳性结果的浓度确定为每ml血清4290份拉沙病毒L RNA，相当于每次反应30份。利用体外转录的RNA、受感染细胞培养物的材料以及来自拉沙热患者和lcmv相关的富营养性肝炎猴子的样本，证明了该检测方法检测各种旧大陆沙粒病毒的能力。L基因PCR检测可能适用于：(i)作为拉沙病毒和LCMV的补充诊断检测；（二）查明怀疑为疾病病原的未知旧大陆沙粒病毒；（iii）筛选未知旧大陆沙粒病毒的潜在宿主。(C) 2007年皇家热带医学与卫生学会。Elsevier Ltd.出版。版权所有。
This study describes an RT-PCR assay targeting the L RNA segment of arenaviruses. Conserved regions were identified in the polymerase domain of the L gene on the basis of published sequences for Lassa virus, lymphocytic choriomeningitis virus (LCMV), Pichinde virus and Tacaribe virus, as well as 15 novel sequences for Lassa virus, LCMV, Ippy virus, Mobala virus and Mopeia virus determined in this study. Using these regions as target sites, a PCR assay for detection of all known Old World arenaviruses was developed and optimized. The concentration that yields 95% positive results in a set of replicate tests (95% detection Limit) was determined to be 4290 copies of Lassa virus L RNA per ml of serum, corresponding to 30 copies per reaction. The ability of the assay to detect various Old World arenaviruses was demonstrated with in vitro transcribed RNA, material from infected cell cultures and samples from patients with Lassa fever and monkeys with LCMV-associated callitrichid hepatitis. The L gene PCR assay may be applicable: (i) as a complementary diagnostic test for Lassa virus and LCMV; (ii) to identify unknown Old World arenaviruses suspected as aetiological agents of disease; and (iii) for screening of potential reservoir hosts for unknown Old World arenaviruses. (C) 2007 Royal Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. All rights reserved.