Rapid identification viruses from nasal pharyngeal aspirates in acute viral respiratory infections by RT-PCR and electrospray ionization mass spectrometry.
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 8h. RT-PCR/ESI-MS demonstrated capacity to detect respiratory viruses identifiable and unidentifiable by conventional methods rapidly.
Johns Hopkins University, Department of Medicine, Baltimore, MD 21205, USA.
Kuan-Fu
KF
Chen
Richard E
RE
Rothman
Padmini
P
Ramachandran
Lawrence
L
Blyn
Rangarajan
R
Sampath
David J
DJ
Ecker
Alexandra
A
Valsamakis
Charlotte A
CA
Gaydos
D006801:Humans; D009305:Nasopharynx / Q000821:virology*; D012141:Respiratory Tract Infections / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014773:Virology / Q000379:methods*; D014777:Virus Diseases / Q000175:diagnosis* / Q000821:virology; D014780:Viruses / Q000145:classification / Q000302:isolation & purification*
PLoS One. 2007 May 30;2(5):e489
17534439
J Infect Dis. 2006 Oct 1;194(7):877-85
16960774
J Clin Microbiol. 2008 Sep;46(9):3056-62
18632902
J Clin Microbiol. 2004 Mar;42(3):1181-4
15004072
J Clin Microbiol. 2009 Mar;47(3):527-32
19129410
Proc Natl Acad Sci U S A. 2005 May 31;102(22):8012-7
15911764
Curr Issues Mol Biol. 2007 Jul;9(2):87-102
17489437
Emerg Infect Dis. 2005 Mar;11(3):373-9
15757550
Clin Lab Med. 2009 Dec;29(4):673-93
19892228
Anal Biochem. 2003 May 1;316(1):50-7
12694726
J Clin Microbiol. 2006 Aug;44(8):2921-32
16891513
Chest. 2003 May;123(5):1664-72
12740288
Clin Microbiol Rev. 2008 Oct;21(4):716-47
18854489
J Clin Microbiol. 2001 Aug;39(8):2779-83
11473991
Clin Chem Lab Med. 2008;46(7):909-18
18605949
Epidemiology. 2005 Sep;16(5):604-12
16135935
J Clin Virol. 2007 Nov;40 Suppl 1:S11-4
18162248
J Clin Microbiol. 2009 Mar;47(3):743-50
19158263
J Clin Virol. 2004 Mar;29(3):179-88
14962787