Serotype-specific detection of enterovirus 71 in clinical specimens by DNA microchip array

被引:35
作者
Shih, SR
Wang, YW
Chen, GW
Chang, LY
Lin, TY
Tseng, MC
Chiang, C
Tsao, KC
Huang, CG
Shio, MR
Tai, JH
Wang, SH
Kuo, RL
Liu, WT
机构
[1] Chang Gung Univ, Sch Med Technol, Taoyuan 333, Taiwan
[2] Chang Gung Mem Hosp, Clin Virol Lab, Dept Clin Pathol, Taoyuan, Taiwan
[3] Dr Chips Biotechnol Inc, Hsinchu, Taiwan
[4] Natl Hlth Res Inst, Div Biostat & Bioinformat, Taipei, Taiwan
[5] Chang Gung Childrens Hosp, Div Pediat Infect Dis, Dept Pediat, Tao Yuan, Taiwan
[6] Natl Yang Ming Univ, Inst Microbiol & Immunol, Taipei 112, Taiwan
[7] Natl Yang Ming Univ, Inst Biotechnol Med, Taipei 112, Taiwan
关键词
microchip; enterovirus; 71;
D O I
10.1016/S0166-0934(03)00151-4
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Enterovirus 71 is an important pathogen that causes high morbidity and mortality in children in Taiwan. Virus isolation in cell cultures has been the standard method for enterovirus 71 identification in Clinical Virology Laboratories. However, virus isolation takes 5-10 days when using cell culture. A microchip for enterovirus 71 detection was developed as an alternative diagnostic method. The novel approach is based on hybridization of amplified DNA specimens with oligonucleotide DNA probes immobilized on a microchip. Two oligonucletides were used as detection probes, the pan-enterovirus sequence located in the 5'-noncoding region (5'-NCR) and the enterovirus 71-specific sequence located in the VP2 region. The diagnostic procedure takes 6 h. One hundred specimens identified as enteroviruses by viral cultures were tested using this microchip, including 67 enterovirus 71 specimens. The sensitivity of the novel method is 89.6% and its specificity is 90.9%. The enterovirus 71-microchip can detect the amplicon derived from viral RNA corresponding to 1-10 virions in a clinical specimen. Microchip array is a potential diagnostic method for identification of enterovirus in the future. (C) 2003 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:55 / 60
页数:6
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