Design and application of 60mer oligonucleotide microarray in SARS coronavirus detection.
Identifieur interne : 006033 ( Main/Exploration ); précédent : 006032; suivant : 006034Design and application of 60mer oligonucleotide microarray in SARS coronavirus detection.
Auteurs : Rong Shi [République populaire de Chine] ; Wenli Ma [République populaire de Chine] ; Qinghua Wu [République populaire de Chine] ; Bao Zhang [République populaire de Chine] ; Yanbin Song [République populaire de Chine] ; Qiuye Guo [République populaire de Chine] ; Weiwei Xiao [République populaire de Chine] ; Yan Wang [République populaire de Chine] ; Wenling Zheng [République populaire de Chine]Source :
- Chinese science bulletin = Kexue tongbao [ 1001-6538 ] ; 2003.
Abstract
The 60mer oligonucleotide microarray was designed and applied to detecting of SARS (severe acute respiratory syndrome) coronavirus. Thirty 60mer specific oligos were designed to cover the whole genome of the first submitted coronavirus strain, according to the sequence of TOR2 (GENEBANK Accession: AY274119). These primers were synthesized and printed into a microarray with 12 ×12 spots. RNAs were extracted from the throat swab and gargling fluid of SARS patients and reverse-transcripted into the double strand cDNAs. The cDNAs were prepared as restricted cDNA fragments by the restriction display (RD) technique and labeled by PCR with the Cy5-universal primer. The labeled samples were then applied to the oligo microarray for hybridization. The diagnostic capability of the microarray was evaluated after the washing and scanning steps. The scanning result showed that samples of SARS patients were hybridized with multiple SARS probes on the microarray, and there is no signal on the negative and blank controls. These results indicate that the genome of SARS coronavirus can be detected in parallel by the 60mer oligonucleotide microarray, which can improve the positive ratio of the diagnosis. The oligo microarray can also be used for monitoring the behavior of the virus genes in different stages of the disease status.
DOI: 10.1007/BF03183928
PubMed: 32214700
Affiliations:
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wicri:level="1"><nlm:affiliation>1Institute of Molecular Biology, First Military Medical University, 510515 Guangzhou, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>1Institute of Molecular Biology, First Military Medical University, 510515 Guangzhou</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author><author><name sortKey="Zheng, Wenling" sort="Zheng, Wenling" uniqKey="Zheng W" first="Wenling" last="Zheng">Wenling Zheng</name><affiliation wicri:level="1"><nlm:affiliation>Department of Medical Research, Guangzhou Liu Hua Qiao Hospital, 510010 Guangzhou, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Medical Research, Guangzhou Liu Hua Qiao Hospital, 510010 Guangzhou</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author></analytic><series><title level="j">Chinese science bulletin = Kexue tongbao</title><idno type="ISSN">1001-6538</idno><imprint><date when="2003" type="published">2003</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The 60mer oligonucleotide microarray was designed and applied to detecting of SARS (severe acute respiratory syndrome) coronavirus. Thirty 60mer specific oligos were designed to cover the whole genome of the first submitted coronavirus strain, according to the sequence of TOR2 (GENEBANK Accession: AY274119). These primers were synthesized and printed into a microarray with 12 ×12 spots. RNAs were extracted from the throat swab and gargling fluid of SARS patients and reverse-transcripted into the double strand cDNAs. The cDNAs were prepared as restricted cDNA fragments by the restriction display (RD) technique and labeled by PCR with the Cy5-universal primer. The labeled samples were then applied to the oligo microarray for hybridization. The diagnostic capability of the microarray was evaluated after the washing and scanning steps. The scanning result showed that samples of SARS patients were hybridized with multiple SARS probes on the microarray, and there is no signal on the negative and blank controls. These results indicate that the genome of SARS coronavirus can be detected in parallel by the 60mer oligonucleotide microarray, which can improve the positive ratio of the diagnosis. The oligo microarray can also be used for monitoring the behavior of the virus genes in different stages of the disease status.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><region><li>Guangdong</li></region><settlement><li>Jiangmen</li></settlement></list><tree><country name="République populaire de Chine"><region name="Guangdong"><name sortKey="Shi, Rong" sort="Shi, Rong" uniqKey="Shi R" first="Rong" last="Shi">Rong Shi</name></region><name sortKey="Guo, Qiuye" sort="Guo, Qiuye" uniqKey="Guo Q" first="Qiuye" last="Guo">Qiuye Guo</name><name sortKey="Ma, Wenli" sort="Ma, Wenli" uniqKey="Ma W" first="Wenli" last="Ma">Wenli Ma</name><name sortKey="Song, Yanbin" sort="Song, Yanbin" uniqKey="Song Y" first="Yanbin" last="Song">Yanbin Song</name><name sortKey="Wang, Yan" sort="Wang, Yan" uniqKey="Wang Y" first="Yan" last="Wang">Yan Wang</name><name sortKey="Wu, Qinghua" sort="Wu, Qinghua" uniqKey="Wu Q" first="Qinghua" last="Wu">Qinghua Wu</name><name sortKey="Xiao, Weiwei" sort="Xiao, Weiwei" uniqKey="Xiao W" first="Weiwei" last="Xiao">Weiwei Xiao</name><name sortKey="Zhang, Bao" sort="Zhang, Bao" uniqKey="Zhang B" first="Bao" last="Zhang">Bao Zhang</name><name sortKey="Zheng, Wenling" sort="Zheng, Wenling" uniqKey="Zheng W" first="Wenling" last="Zheng">Wenling Zheng</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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