[KSCI] Korea Science Citation Index Service

http://dx.doi.org/10.5352/JLS.2008.18.3.374

Comparison of the Real-Time Nucleic Acid Sequence-Based Amplification (NASBA) Assay, Reverse Transcription-PCR (RT-PCR) and Virus Isolation for the Detection of Enterovirus RNA.

Na, Young-Ran (Division of Epidemiology, Busan Institute of Health and Environment)
Joe, Hyeon-Cheol (Division of Epidemiology, Busan Institute of Health and Environment)
Lee, Young-Suk (Division of Epidemiology, Busan Institute of Health and Environment)
Bin, Jae-Hun (Division of Epidemiology, Busan Institute of Health and Environment)
Cheigh, Hong-Sik (Division of Epidemiology, Busan Institute of Health and Environment)
Min, Sang-Kee (Division of Epidemiology, Busan Institute of Health and Environment)

Publication Information

Journal of Life Science / v.18, no.3, 2008 , pp. 374-380 More about this Journal

Abstract

Rapid detection of enterovirus (EVs) is important in the management of aseptic meningitis. We examined the relative efficiency and specificity of the real-time nucleic acid sequence-based amplification (NASBA) comparing with the established reverse transcription polymerase chain reaction (RT-PCR) and viral culture method which were used for the detection of enterovirus RNA in clinical specimens. Of the total 292 samples, 145 were found to be positive to enterovirus RNA by real-time NASBA, 101 were positive by viral culture, and 86 were positive by RT-PCR. 147 samples and 46 samples were determined to be negative and positive by all methods respectively, but 4 samples were positive only by real-time NASBA. To compare the specificity of each method, various clinical samples which were diagnosed for herpes simplex virus (HSV)-1, HSV-2, adenovirus, mumps, and rhinovirus were applied. Except one rhinovirus sample which was false positive to enterovirus RNA by RT-PCR, the other different samples were negative to all three methods. The real-time NASBA procedure can be completed within 5 hours in contrast with 9 hours for the RT-PCR and 3-14 days for the viral culture. From this study, it was suggested that the real-time NASBA assay could be a standardized, rapid, specific, and sensitive procedure for the detection of enterovirus RNA.

Keywords

Enterovirus; real-time NASBA; RT-PCR; viral culture;

Citations & Related Records

Reference

1	Gorgioevski-Hrisoho, M., J. D. Schmacher, N. Vilimonovic, D. Germann and L. Matter. 1998. Detection by PCR of enteroviruses in cerebrospinal fluid during a summer outbreak of aseptic meningitis in Switzerland. J. Clin. Microbiol. 36, 2408-2412.
2	Hellen, C. U. and E. Wimmer. 1995. Enterovirus structure and assembly. pp. 155-174, Human enterovirus infection, ASM press, Washington DC.
3	Grandien, M., M. Forsgren and A. Ehrnst. 1995. Enteroviruses diagnostic procedures for viral, rickettsial and chlamydial infection. pp. 279-297, In Lennette E. H. and E. T. Lennette, 7th eds. American Public Health Association, Washington DC.
4	Capaul, S. E. and M. H. Gorgioevski. 2005. Detection of enterovirus RNA in cerebrospinal fluid (CSF) using NucliSens EasyQ enterovirus assay. J. Clin. Virol. 32, 236-240. DOI ScienceOn
5	Compton, J. 1991. Nucleic acid sequence based amplification. Nature 350, 91. DOI ScienceOn
6	Fox, J. D., S. Han, A. Samuelson, Y. Zhang, M. L. Neale and D. Westmoreland. 2002. Development and evaluation of a nucleic acid sequence based amplification (NASBA) for diagnosis of enterovirus infections using the NucliSens Basic kit. J. Clinical Virol. 24, 117-120. DOI ScienceOn
7	Loens, K., D. Ursi, M. Ieven, A. P. Van, P. Sillekens, P. Oudshoorn and H. Goossens. 2002. Detection of Mycoplasma pneumoniae in spiked clinical samples by Nucleic Acid Sequence-Based Amplification. J. Clin. Microbiol. 40, 1339-1345. DOI ScienceOn
8	Landry, M. L., R. Garner and D. Ferguson. 2003. Rapid enterovirus RNA detection in clinical specimens by using nucleic acid sequence based amplification. J. Clin. Microbiol. 41, 346-350. DOI ScienceOn
9	Min, J. and A. J. Baeumer. 2002. Highly sensitive and specific detection of viable Escherichia coli in drinking water. Anal. Biochem. 303, 186-193. DOI ScienceOn
10	Lai, K. K. Y., L. Cook, S. Wendt, L. Conrey and K. R. Jerome. 2003. Evaluation of realtime PCR versus PCR with liquid phase hybridization for detection of enterovirus RNA in cerebrospinal fluid. J. Clin. Microbiol. 41, 3133-3141. DOI ScienceOn
11	Landry, M. L., R. Garner and D. Ferguson. 2003. Comparison of the NuceiSens basic kit (nucleic acid sequence based amplification) and Argene Biosoft enterovirus consensus reverse transcription PCR assay for rapid enterovirus RNA in clinical specimens. J. Clin. Microbiol. 41, 5006-5010. DOI ScienceOn
12	Zoll, G. J., W. J. Melcher, H. Kopecka, G. Jambros and H. J. Vanderpoel. 1992. General primer mediated polymeric chain reaction for detection of enteroviruses; application for diagnostic routine and persistant infections. J. Clin. Microbiol. 30, 160.
13	Rotbart, H. A. 1990. Enzymatic RNA amplification of enteroviruses. J. Clin. Microbiol. 28, 438-442.
14	Wacharapluesadee, S. and T. L. Hemachudha. 2001. Nucleic-acid sequence based amplification in the rapid diagnosis of rabies. J. Clin. Microbiol. 358, 892-893.
15	Oberste, M. S., K. Maher, D. R. Kilpatrick, M. R. Flemister, B. A. Brown and M. A. Pallansch. 1999. Typing of human enteroviruses by partial sequencing of VP1. J. Clin. Microbiol. 37, 1288-1293.
16	Oberste, M. S., K. Maher, D. R. Kilpatrick and A. Pallansch. 1999. Molecular evolution of the human enteroviruses: Correlation of serotype with VP1 sequence and application to picornavirus classification. J. Virol. 73, 1941-1948.
17	Heim, A. and J. Schumann. 2002. Development and evaluation of a nucleic acid sequence based amplification (NASBA) protocol for the detection of enterovirus RNA in cerebrospinal fluid samples. J. Virological Methods 103, 101-107. DOI ScienceOn
18	Hsiung, G. D. 1994. Picornaviridae. pp. 119, 4th eds. Hsiung's diagnostic virology, New Haven, Yale University press.
19	Ginocchion, C., F. Zhang, A. Malhotra, R. Manji, P. Sillekens, F. Helma, M. Overdyk and M. Peeters. 2005. Development technical performance and clinical evaluation of a nucli sens basic kit application for detection of enterovirus RNA cerebrospinal fluid. J. Clin. Microbiol. 2616-2623.
20	Glimarker, M., B. Johasson, P. Olcen, A. Ehrnst and M. Forsgren. 1993. Detection of enterovirus RNA by polymeric chain reaction in cerebrospinal fluid from patients with aseptic meningitis. Scan. J. Infect. Dis. 25, 547-557. DOI ScienceOn
21	Jean, J., B. Blais, A. Darveau and I. Fliss. 2001. Detection of hepatitis a virus by the nucleic acid sequence-based amplification technique and comparison with reverse transcription- PCR. Appl. Environ. Microbiol. 67, 5593-5600. DOI ScienceOn