Evaluation of Three Automated Nucleic Acid Extraction Systems for Identification of Respiratory Viruses in Clinical Specimens by Multiplex Real-Time PCR

被引:10
作者
Kim, Yoonjung [1 ]
Han, Mi-Soon [1 ]
Kim, Juwon [2 ]
Kwon, Aerin [3 ]
Lee, Kyung-A [1 ]
机构
[1] Yonsei Univ, Coll Med, Dept Lab Med, Seoul 135720, South Korea
[2] Yonsei Univ, Wonju Coll Med, Dept Lab Med, Wonju, South Korea
[3] Green Cross Labs, Yongin, South Korea
关键词
RT-PCR; SEEPLEX RV15; NUCLISENS EASYMAG; CULTURE; RNA; INFLUENZA; ASSAYS; KIT; DNA;
D O I
10.1155/2014/430650
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
A total of 84 nasopharyngeal swab specimens were collected from 84 patients. Viral nucleic acid was extracted by three automated extraction systems: QIAcube (Qiagen, Germany), EZ1 Advanced XL (Qiagen), and MICROLAB Nimbus IVD (Hamilton, USA). Fourteen RNA viruses and two DNA viruses were detected using the Anyplex II RV16 Detection kit (Seegene, Republic of Korea). The EZ1 Advanced XL system demonstrated the best analytical sensitivity for all the three viral strains. The nucleic acids extracted by EZ1 Advanced XL showed higher positive rates for virus detection than the others. Meanwhile, the MICROLAB Nimbus IVD system was comprised of fully automated steps from nucleic extraction to PCR setup function that could reduce human errors. For the nucleic acids recovered from nasopharyngeal swab specimens, the QIAcube system showed the fewest false negative results and the best concordance rate, and it may be more suitable for detecting various viruses including RNA and DNA virus strains. Each system showed different sensitivity and specificity for detection of certain viral pathogens and demonstrated different characteristics such as turnaround time and sample capacity. Therefore, these factors should be considered when new nucleic acid extraction systems are introduced to the laboratory.
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页数:8
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共 23 条
[1]   Development of three multiplex RT-PCR assays for the detection of 12 respiratory RNA viruses [J].
Bellau-Pujol, S ;
Vabret, A ;
Legrand, L ;
Dina, J ;
Gouarin, S ;
Petitjean-Lecherbonnier, J ;
Pozzetto, B ;
Ginevra, C ;
Freymuth, F .
JOURNAL OF VIROLOGICAL METHODS, 2005, 126 (1-2) :53-63
[2]   Ratification vote on taxonomic proposals to the International Committee on Taxonomy of Viruses (2009) [J].
Carstens, E. B. .
ARCHIVES OF VIROLOGY, 2010, 155 (01) :133-146
[3]   Comparison of the NucliSens easyMAG and Qiagen BioRobot 9604 nucleic acid extraction systems for detection of RNA and DNA respiratory viruses in nasopharyngeal aspirate samples [J].
Chan, Kwok Hung ;
Yam, Wing Cheong ;
Pang, Chiu Mei ;
Chan, Kit Man ;
Lam, Siu Yan ;
Lo, Kam Fai ;
Poon, Leo L. M. ;
Peiris, J. S. Malik .
JOURNAL OF CLINICAL MICROBIOLOGY, 2008, 46 (07) :2195-2199
[4]   Evaluation of the AdvanSure™ real-time RT-PCR compared with culture and Seeplex RV15 for simultaneous detection of respiratory viruses [J].
Cho, Chi Hyun ;
Lee, Chang Kyu ;
Nam, Myung-Hyun ;
Yoon, Soo-Young ;
Lim, Chae Seung ;
Cho, Yunjung ;
Kim, Young Kee .
DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE, 2014, 79 (01) :14-18
[5]   Evaluation of a novel real-time RT-PCR using TOCE technology compared with culture and Seeplex RV15 for simultaneous detection of respiratory viruses [J].
Cho, Chi Hyun ;
Chulten, Bayarjavkhlan ;
Lee, Chang Kyu ;
Nam, Myung Hyun ;
Yoon, Soo Young ;
Lim, Chae Seung ;
Cho, Yunjung ;
Kim, Young Kee .
JOURNAL OF CLINICAL VIROLOGY, 2013, 57 (04) :338-342
[6]   Viral Infection in Patients with Severe Pneumonia Requiring Intensive Care Unit Admission [J].
Choi, Sang-Ho ;
Hong, Sang-Bum ;
Ko, Gwang-Beom ;
Lee, Yumi ;
Park, Hyun Jung ;
Park, So-Youn ;
Moon, Song Mi ;
Cho, Oh-Hyun ;
Park, Ki-Ho ;
Chong, Yong Pil ;
Kim, Sung-Han ;
Huh, Jin Won ;
Sung, Heungsup ;
Do, Kyung-Hyun ;
Lee, Sang-Oh ;
Kim, Mi-Na ;
Jeong, Jin-Yong ;
Lim, Chae-Man ;
Kim, Yang Soo ;
Woo, Jun Hee ;
Koh, Younsuck .
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE, 2012, 186 (04) :325-332
[7]   Optimization of one-step duplex real-time RT-PCR for detection of influenza and respiratory syncytial virus in nasopharyngeal aspirates [J].
de-Paris, Fernanda ;
Beck, Caroline ;
Mombach Pinheiro Machado, Alice Beatriz ;
Paiva, Rodrigo Minuto ;
Menezes, Denise da Silva ;
Nunes, Luciana de Souza ;
Kuchenbecker, Ricardo ;
Barth, Afonso Luis .
JOURNAL OF VIROLOGICAL METHODS, 2012, 186 (1-2) :189-192
[8]   Emerging pathogens: Challenges and successes of molecular diagnostics [J].
Dong, Jianli ;
Olano, Juan P. ;
McBride, Jere W. ;
Walker, David H. .
JOURNAL OF MOLECULAR DIAGNOSTICS, 2008, 10 (03) :185-197
[9]   Epidemiology of viral respiratory tract infections in a prospective cohort of infants and toddlers attending daycare [J].
Fairchok, Mary P. ;
Martin, Emily T. ;
Chambers, Susan ;
Kuypers, Jane ;
Behrens, Melinda ;
Braun, LoRanee E. ;
Englund, Janet A. .
JOURNAL OF CLINICAL VIROLOGY, 2010, 49 (01) :16-20
[10]   Nucleic acid amplification tests for detection of respiratory viruses [J].
Fox, Julie D. .
JOURNAL OF CLINICAL VIROLOGY, 2007, 40 :S15-S23