Rapid detection and differentiation of human noroviruses using RT-PCR coupled to electrospray ionization mass spectrometry

被引：2
|
作者：
Hellberg, Rosalee S. [1 ]
Li, Feng [2 ]
Sampath, Rangarajan [2 ]
Yasuda, Irene J. [2 ]
Carolan, Heather E. [2 ]
Wolfe, Julia M. [3 ]
Brown, Michael K. [3 ]
Alexander, Richard C. [3 ]
Williams-Hill, Donna M. [4 ]
Martin, William B. [4 ]
机构：
[1] Chapman Univ, Schmid Coll Sci & Technol, Orange, CA 92866 USA
[2] Abbott, Ibis Biosci, Carlsbad, CA 92008 USA
[3] Orange Cty Publ Hlth Lab, Santa Ana, CA 92706 USA
[4] USDA, Off Regulatory Affairs, Pacific Reg Lab Southwest, Irvine, CA 92612 USA
来源：
FOOD MICROBIOLOGY | 2014年 / 44卷
关键词：
Norovirus; Hepatitis A virus; RT-PCR; Electrospray ionization mass spectrometry; Genotyping; UNITED-STATES; ESCHERICHIA-COLI; IDENTIFICATION; OUTBREAKS; EPIDEMIOLOGY; STRAIN; GASTROENTERITIS; SURVEILLANCE; INFECTION; PATHOGENS;
D O I：
10.1016/j.fm.2014.05.017
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
The goal of this study was to develop an assay for the detection and differentiation of noroviruses using RT-PCR followed by electrospray ionization mass spectrometry (ESI-MS). Detection of hepatitis A virus was also considered. Thirteen primer pairs were designed for use in this assay and a reference database was created using GenBank sequences and reference norovirus samples. The assay was tested for inclusivity and exclusivity using 160 clinical norovirus samples, 3 samples of hepatitis A virus and 3 other closely related viral strains. Results showed that the assay was able to detect norovirus with a sensitivity of 92% and a specificity of 100%. Norovirus identification at the genogroup level was correct for 98% of samples detected by the assay and for 75% of a subset of samples (n = 32) compared at the genotype level. Identification of norovirus genotypes is expected to improve as more reference samples are added to the database. The assay was also capable of detecting and genotyping hepatitis A virus in all 3 samples tested. Overall, the assay developed here allows for detection and differentiation of noroviruses within one working day and may be used as a tool in surveillance efforts or outbreak investigations. (C) 2014 Elsevier Ltd. All rights reserved.
引用
收藏
页码：71 / 80
页数：10
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