A universal microarray for detection of SARS coronavirus.
Severe acute respiratory syndrome (SARS) is caused by the SARS coronavirus (SARS-CoV). There are many point mutations among SARS-CoV genome sequences. Previous studies suggested that the mutations are correlated closely with the SARS epidemic. It was found that the bases of six nucleotide positions (nt9404, nt9479, nt19838, nt21721, nt22222 and nt27827) with high-mutation rate have an important relationship with the SARS epidemic. For viral detection as well as genotyping, a universal microarray system was developed that combines RT-PCR and ligase detection reaction (LDR). The Zip Codes attached covalently to a slide remain constant and their complementary Zip Codes (cZip Codes) can be used for tagging target sequence, making the microarrays universal. The discriminating oligonucleotides contain on the 5' end "cZip Codes" that are used to direct LDR product to specific Zip Codes attached covalently to a slide. Since Zip Codes have no homology to either the target sequence or to other sequences in the genomes of both human host and SARS-CoV, there was no false signal due to mismatch hybridizations. 20 samples assayed with the universal microarray were confirmed by DNA sequencing, demonstrating that this microarray system is a promising diagnostic tool for detection and genotyping of the SARS-CoV.
Long
Wei-Hong
WH
School of Life Science and Technology, Shanghai JiaoTong University, 1954 Hua-Shan Road, Shanghai 200030, China.
Xiao
Hua-Sheng
HS
Gu
Xiao-Mei
XM
Zhang
Qing-Hua
QH
Yang
Hong-Jun
HJ
Zhao
Guo-Ping
GP
Liu
Jian-Hua
JH
D019943:Amino Acid Substitution; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D022521:Ligase Chain Reaction; D025202:Molecular Diagnostic Techniques; D017720:Molecular Epidemiology; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D017354:Point Mutation; D020133:Reverse Transcriptase Polymerase Chain Reaction; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000302:isolation & purification*; D045169:Severe Acute Respiratory Syndrome / Q000175:diagnosis / Q000821:virology*