Development of loop-mediated isothermal amplification assay for detection of human coronavirus-NL63

被引：40
作者：
Pyrc, Krzysztof [1 ]
Milewska, Aleksandra [1 ]
Potempa, Jan [1 ,2 ]
机构：
[1] Jagiellonian Univ, Fac Biochem Biophys & Biotechnol, Dept Microbiol, PL-30387 Krakow, Poland
[2] Univ Louisville, Sch Dent, Oral Hlth & Syst Dis Res Grp, Louisville, KY 40292 USA
来源：
JOURNAL OF VIROLOGICAL METHODS | 2011年 / 175卷 / 01期
关键词：
Coronavirus; HCoV-NL63; LAMP; Loop-mediated isothermal amplification; RESPIRATORY-TRACT INFECTIONS; ACUTE KAWASAKI-DISEASE; NL63; INFECTION; CHILDREN; ASSOCIATION; VIRUS; CROUP; IDENTIFICATION; HKU1;
D O I：
10.1016/j.jviromet.2011.04.024
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Human coronavirus NL63 was identified in 2004 in the Netherlands. Due to the high prevalence and world-wide distribution of this pathogen, it is essential to develop a sensitive and specific detection assay suitable for use in a routine diagnostic laboratory. Techniques based on PCR or real-time PCR are laborious and expensive. Detailed analysis of the HCoV-NL63 genome permitted the identification of a conserved nucleic acid sequential motif, which was sufficient for the design of a loop-mediated isothermal amplification (LAMP) assay. Evaluation of the method showed that the test is specific to HCoV-NL63 and that it does not cross-react with other respiratory viruses. The detection limit was found to be 1 copy of RNA template per reaction in cell culture supernatants and clinical specimens. (C) 2011 Elsevier B.V. All rights reserved.
引用
收藏
页码：133 / 136
页数：4
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