Rapid and Specific Detection of All Known Nipah virus Strains' Sequences With Reverse Transcription-Loop-Mediated Isothermal Amplification
文献类型：期刊论文
| 作者 | Ma, Liping1,2,3; Chen, Zhen4; Guan, Wuxiang4; Chen, Quanjiao1; Liu, Di2,3 |
| 刊名 | FRONTIERS IN MICROBIOLOGY
 |
| 出版日期 | 2019-03-11 |
| 卷号 | 10页码:10 |
| ISSN号 | 1664-302X |
| 关键词 | Nipah virus reverse transcription-loop-mediated isothermal amplification RT-LAMP rapid detection N gene |
| DOI | 10.3389/fmicb.2019.00418 |
| 英文摘要 | Nipah virus (NiV) is a zoonotic virus and can be transmitted through contaminated food or directly between people. NiV is classified as a Biosafety Level 4 agent, not only because of its relatively high case fatality rate, but also because there is no vaccine or other medical countermeasures and it appears to be transmitted by fomites/particulates. The development of rapid detection assay for NiV is of great importance because no effective field test is currently available. In this study, an isothermal (65 degrees C) reverse transcription-loop-mediated isothermal amplification (RT-LAMP) method was developed, targeting the nucleocapsid protein (N) gene, for the rapid detection of NiV, and was compared with conventional RT-PCR. Three pseudoviruses of NiV N gene representing all known strains were constructed to replace live NiV. A set of RT-LAMP primers, targeting a highly conserved region of the N gene in the viral genome was designed to identify all known NiV strains. Sensitivity tests indicated that the detection limit of the RT-LAMP assay was approximately 100 pg of total NiV pseudovirus RNA, which is at least 10-fold higher than that of conventional RT-PCR. Specificity tests showed that there was no cross-reactivity with nucleocapsid protein gene of Hendra virus, Newcastle disease virus, Japanese encephalitis virus, or Influenza A virus. The RT-LAMP assay provides results within 45 min, and requires no sophisticated instruments, except an isothermal water bath or metal bath with 1 mu l calcein indicator. An analysis of the clinical samples showed that the assay had good stability. In conclusion, systematic experiments have shown that the RT-LAMP assay developed here effectively detects three NiV pseudoviruses representing all known strains of NiV, with high specificity, sensitivity and stability. |
| 资助项目 | Ministry of Science and Technology of the People's Republic of China Key Research and Development Program[2016YFC1200805] ; Advanced Customer Cultivation Project of Wuhan National Biosafety Laboratory, Chinese Academy of Sciences ; National Science and Technology Major Project[2018ZX10101004] |
| WOS研究方向 | Microbiology |
| 语种 | 英语 |
| 出版者 | FRONTIERS MEDIA SA |
| WOS记录号 | WOS:000460774900001 |
| 源URL | [http://202.127.146.157/handle/2RYDP1HH/6964]  |
| 专题 | 中国科学院武汉植物园 |
| 通讯作者 | Chen, Quanjiao; Liu, Di |
| 作者单位 | 1.Chinese Acad Sci, Wuhan Inst Virol, CAS Key Lab Special Pathogens & Biosafety, Wuhan, Hubei, Peoples R China 2.Chinese Acad Sci, Computat Virol Grp, Wuhan Inst Virol, Wuhan, Hubei, Peoples R China 3.Univ Chinese Acad Sci, Beijing, Peoples R China 4.Chinese Acad Sci, Ctr Emerging Infect Dis, Wuhan Inst Virol, Wuhan, Hubei, Peoples R China |
| 推荐引用方式 GB/T 7714 | Ma, Liping,Chen, Zhen,Guan, Wuxiang,et al. Rapid and Specific Detection of All Known Nipah virus Strains' Sequences With Reverse Transcription-Loop-Mediated Isothermal Amplification[J]. FRONTIERS IN MICROBIOLOGY,2019,10:10. |
| APA | Ma, Liping,Chen, Zhen,Guan, Wuxiang,Chen, Quanjiao,&Liu, Di.(2019).Rapid and Specific Detection of All Known Nipah virus Strains' Sequences With Reverse Transcription-Loop-Mediated Isothermal Amplification.FRONTIERS IN MICROBIOLOGY,10,10. |
| MLA | Ma, Liping,et al."Rapid and Specific Detection of All Known Nipah virus Strains' Sequences With Reverse Transcription-Loop-Mediated Isothermal Amplification".FRONTIERS IN MICROBIOLOGY 10(2019):10. |
入库方式： OAI收割
来源：武汉植物园
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