中国科学院机构知识库网格
Chinese Academy of Sciences Institutional Repositories Grid
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

AuthorMa, Liping1,2,3; Chen, Zhen4; Guan, Wuxiang4; Chen, Quanjiao1; Liu, Di2,3
SourceFrontiers in microbiology
Issued Date2019-03-11
Volume10Pages:10
ISSN1664-302X
KeywordNipah virus Reverse transcription-loop-mediated isothermal amplification Rt-lamp Rapid detection N gene
DOI10.3389/fmicb.2019.00418
Corresponding AuthorChen, quanjiao(chenqj@wh.iov.cn) ; Liu, di(liud@wh.iov.cn)
English AbstractNipah 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.
WOS KeywordHENDRA VIRUS ; ENCEPHALITIS ; BANGLADESH ; TRANSMISSION ; INFECTION ; DIAGNOSIS ; ASSAY ; LAMP
WOS Research AreaMicrobiology
WOS SubjectMicrobiology
Language英语
WOS IDWOS:000460774900001
PublisherFRONTIERS MEDIA SA
URIhttp://www.irgrid.ac.cn/handle/1471x/2373173
Collection武汉病毒研究所
Corresponding AuthorChen, Quanjiao; Liu, Di
Affiliation1.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
Recommended Citation
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.

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来源:武汉病毒研究所

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