Rapid identification viruses from nasal pharyngeal aspirates in acute viral respiratory infections by RT-PCR and electrospray ionization mass spectrometry

被引:35
作者
Chen, Kuan-Fu [1 ,2 ,3 ]
Rothman, Richard E. [1 ]
Ramachandran, Padmini [1 ]
Blyn, Lawrence [4 ]
Sampath, Rangarajan [4 ]
Ecker, David J. [4 ]
Valsamakis, Alexandra [1 ]
Gaydos, Charlotte A. [1 ]
机构
[1] Johns Hopkins Univ, Div Infect Dis, Dept Med, Baltimore, MD 21205 USA
[2] Chang Gung Univ, Tao Yuan, Taiwan
[3] Chang Gung Mem Hosp, Keelung, Taiwan
[4] Ibis Biosci Inc, Carlsbad, CA USA
关键词
Electrospray ionization mass spectrometry; Respiratory tract infections; Virus; Diagnosis; Surveillance; REVERSE TRANSCRIPTION-PCR; ACID AMPLIFICATION TESTS; DIAGNOSIS; ASSAYS; PATHOGENS;
D O I
10.1016/j.jviromet.2011.01.007
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Diagnosis of the etiologic agent of respiratory viral infection relies traditionally on culture or antigen detection. This pilot evaluation compared performance characteristics of the RT-PCR and electrospray ionization mass spectrometry (RT-PCR/ESI-MS) platform to conventional virologic methods for identifying multiple clinically relevant respiratory viruses in nasopharyngeal aspirates. The RT-PCR/ESI-MS respiratory virus surveillance kit was designed to detect respiratory syncytial virus, influenza A and B. parainfluenza types 1-4, adenoviridae types A-F, coronaviridae, human bocavirus, and human metapneumovirus. Patients (N = 192) attending an emergency department during the 2007-2008 respiratory season consented, and "excess" frozen archived nasopharyngeal aspirates were analysed; 46 were positive by conventional virology and 69 by RT-PCR/ESI-MS, among which there were six samples with multiple viral pathogens detected. The sensitivity and specificity of the assay were 89.1% and 80.3%, respectively. Additional viruses that were not identified by conventional virology assays were detected (4 human bocaviruses and 7 coronaviruses). Samples in which the RT-PCR/ESI-MS results disagreed with conventional virology were sent for analysis by a third method using a commercial RT-PCR-based assay, which can identify viruses not detectable by conventional virologic procedures. Time to first result of RT-PCR/ESI-MS was 8 h. RT-PCR/ESI-MS demonstrated capacity to detect respiratory viruses identifiable and unidentifiable by conventional methods rapidly. (C) 2011 Elsevier B.V. All rights reserved.
引用
收藏
页码:60 / 66
页数:7
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