Serotype-specific detection of enterovirus 71 in clinical specimens by DNA microchip array.
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 oligonucleotides 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.
Shih
Shin-Ru
SR
School of Medical Technology, Chang Gung University, 259 Wen-Hua 1st Road, Kwei-Shan, Tao-Yuan 333, Taiwan, ROC. srshih@mail.cgu.edu.tw
Wang
Yih-Weng
YW
Chen
Guang-Wu
GW
Chang
Luan-Yin
LY
Lin
Tzou-Yien
TY
Tseng
Mei-Chen
MC
Chiang
Chiayn
C
Tsao
Kuo-Chien
KC
Huang
Chung Guei
CG
Shio
Mei-Ren
MR
Tai
Jui-Hung
JH
Wang
Shin-Hwan
SH
Kuo
Rei-Lin
RL
Liu
Wu-Tse
WT
D020121:5' Untranslated Regions / Q000235:genetics; D017931:DNA Primers; D018076:DNA, Complementary / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification*; D004769:Enterovirus Infections / Q000175:diagnosis; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D012680:Sensitivity and Specificity; D012703:Serotyping; D014764:Viral Proteins / Q000235:genetics
D020121:5' Untranslated Regions; D017931:DNA Primers; D018076:DNA, Complementary; D004279:DNA, Viral; D014764:Viral Proteins