One-tube nested RT-PCR enabled by using a plastic film and its application for the rapid detection of SARS-virus
Identifieur interne : 000804 ( PascalFrancis/Checkpoint ); précédent : 000803; suivant : 000805One-tube nested RT-PCR enabled by using a plastic film and its application for the rapid detection of SARS-virus
Auteurs : Sheng-Ce Tao [République populaire de Chine] ; DI JIANG [République populaire de Chine] ; Hong-Li Lu [République populaire de Chine] ; Wan-Li Xing [République populaire de Chine] ; Yu-Xiang Zhou [République populaire de Chine] ; JING CHENG [République populaire de Chine]Source :
- Biotechnology letters [ 0141-5492 ] ; 2004.
Descripteurs français
- Pascal (Inist)
English descriptors
Abstract
A general strategy based on the use of a plastic film that allows reverse transcription and nested-PCR in a single closed-tube has been developed. The reaction mixture for the second PCR amplification is quarantined in the cap of the reaction tube during the first round amplification by a piece of plastic film, and later introduced into the PCR amplicons from the first round reaction by centrifugation without opening the reaction tube. The main advantages of our method are its high sensitivity, specificity, simplicity, cost effectiveness, low risk of contamination and the ease in establishment of conditions for nested-PCR. The method has been successfully applied to the detection of the genomic RNA from SARS-CoV, the detection limit of this method is comparable to that of the conventional two-tube nested PCR system. This method could be easily adapted to the detection of other targets.
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<front><div type="abstract" xml:lang="en">A general strategy based on the use of a plastic film that allows reverse transcription and nested-PCR in a single closed-tube has been developed. The reaction mixture for the second PCR amplification is quarantined in the cap of the reaction tube during the first round amplification by a piece of plastic film, and later introduced into the PCR amplicons from the first round reaction by centrifugation without opening the reaction tube. The main advantages of our method are its high sensitivity, specificity, simplicity, cost effectiveness, low risk of contamination and the ease in establishment of conditions for nested-PCR. The method has been successfully applied to the detection of the genomic RNA from SARS-CoV, the detection limit of this method is comparable to that of the conventional two-tube nested PCR system. This method could be easily adapted to the detection of other targets.</div>
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<s5>10</s5>
</fC03>
<fC03 i1="03" i2="X" l="ENG"><s0>Rapid technique</s0>
<s5>10</s5>
</fC03>
<fC03 i1="03" i2="X" l="SPA"><s0>Técnica rápida</s0>
<s5>10</s5>
</fC03>
<fC03 i1="04" i2="X" l="FRE"><s0>Détection</s0>
<s5>11</s5>
</fC03>
<fC03 i1="04" i2="X" l="ENG"><s0>Detection</s0>
<s5>11</s5>
</fC03>
<fC03 i1="04" i2="X" l="SPA"><s0>Detección</s0>
<s5>11</s5>
</fC03>
<fC03 i1="05" i2="X" l="FRE"><s0>Virus</s0>
<s2>NW</s2>
<s5>13</s5>
</fC03>
<fC03 i1="05" i2="X" l="ENG"><s0>Virus</s0>
<s2>NW</s2>
<s5>13</s5>
</fC03>
<fC03 i1="05" i2="X" l="SPA"><s0>Virus</s0>
<s2>NW</s2>
<s5>13</s5>
</fC03>
<fN21><s1>166</s1>
</fN21>
<fN44 i1="01"><s1>OTO</s1>
</fN44>
<fN82><s1>OTO</s1>
</fN82>
</pA>
</standard>
</inist>
<affiliations><list><country><li>République populaire de Chine</li>
</country>
<settlement><li>Pékin</li>
</settlement>
</list>
<tree><country name="République populaire de Chine"><noRegion><name sortKey="Tao, Sheng Ce" sort="Tao, Sheng Ce" uniqKey="Tao S" first="Sheng-Ce" last="Tao">Sheng-Ce Tao</name>
</noRegion>
<name sortKey="Di Jiang" sort="Di Jiang" uniqKey="Di Jiang" last="Di Jiang">DI JIANG</name>
<name sortKey="Jing Cheng" sort="Jing Cheng" uniqKey="Jing Cheng" last="Jing Cheng">JING CHENG</name>
<name sortKey="Jing Cheng" sort="Jing Cheng" uniqKey="Jing Cheng" last="Jing Cheng">JING CHENG</name>
<name sortKey="Jing Cheng" sort="Jing Cheng" uniqKey="Jing Cheng" last="Jing Cheng">JING CHENG</name>
<name sortKey="Lu, Hong Li" sort="Lu, Hong Li" uniqKey="Lu H" first="Hong-Li" last="Lu">Hong-Li Lu</name>
<name sortKey="Tao, Sheng Ce" sort="Tao, Sheng Ce" uniqKey="Tao S" first="Sheng-Ce" last="Tao">Sheng-Ce Tao</name>
<name sortKey="Xing, Wan Li" sort="Xing, Wan Li" uniqKey="Xing W" first="Wan-Li" last="Xing">Wan-Li Xing</name>
<name sortKey="Xing, Wan Li" sort="Xing, Wan Li" uniqKey="Xing W" first="Wan-Li" last="Xing">Wan-Li Xing</name>
<name sortKey="Zhou, Yu Xiang" sort="Zhou, Yu Xiang" uniqKey="Zhou Y" first="Yu-Xiang" last="Zhou">Yu-Xiang Zhou</name>
<name sortKey="Zhou, Yu Xiang" sort="Zhou, Yu Xiang" uniqKey="Zhou Y" first="Yu-Xiang" last="Zhou">Yu-Xiang Zhou</name>
<name sortKey="Zhou, Yu Xiang" sort="Zhou, Yu Xiang" uniqKey="Zhou Y" first="Yu-Xiang" last="Zhou">Yu-Xiang Zhou</name>
</country>
</tree>
</affiliations>
</record>
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