CovidV2, Main, Exploration, bibRecord, 001364

Development and application of one-step multiplex reverse transcription PCR for simultaneous detection of five diarrheal viruses in adult cattle

Identifieur interne : 001364 ( Main/Exploration ); précédent : 001363; suivant : 001365

Development and application of one-step multiplex reverse transcription PCR for simultaneous detection of five diarrheal viruses in adult cattle

Auteurs : Masaharu Fukuda [Japon] ; Kazufumi Kuga [Japon] ; Ayako Miyazaki [Japon] ; Tohru Suzuki [Japon] ; Keito Tasei [Japon] ; Tsunehiko Aita [Japon] ; Masaji Mase [Japon] ; Makoto Sugiyama [Japon] ; Hiroshi Tsunemitsu [Japon]

Source :

Archives of virology [ 0304-8608 ] ; 2012.

RBID : Pascal:12-0254670

Descripteurs français

Pascal (Inist)
- Bovin, Réaction chaîne polymérase multiplex, Réaction chaîne polymérase RT, Détection, Diarrhée, Vétérinaire.
Wicri :
- topic : Bovin, Vétérinaire.

English descriptors

KwdEn :
- Bovine, Detection, Diarrhea, Multiplex polymerase chain reaction, Reverse transcription polymerase chain reaction, Veterinary.

Abstract

A one-step multiplex reverse transcription (RT)-PCR method was developed for the simultaneous detection of five viruses causing diarrhea in adult cattle: bovine group A rotavirus (GAR), bovine group B rotavirus (GBR), bovine group C rotavirus (GCR), bovine corona-virus (BCV), and bovine torovirus (BToV). The detection limit of the one-step multiplex RT-PCR for GAR, GCR, BCV, and BToV was 10², 10°, 10¹, and 10² TCID₅₀/ml, respectively, and that for GBR was 10⁶ copies/ml. The one-step multiplex RT-PCR with newly designed primers to detect GAR had higher sensitivity than a single RT-PCR with conventional primers, with no false-positive reactions observed for ten other kinds of bovine RNA viruses To assess its field applicability, 59 of 60 fecal samples containing one of these five viruses from all 25 epidemic diarrhea outbreaks in adult cattle were positive in the one-step multiplex RT-PCR assay. Furthermore, using four additional fecal samples containing two viruses (GBR and BCV or BToV), two amplified products of the expected sizes were obtained simultaneously. In contrast, all 80 fecal samples lacking the five target viruses from normal adult cattle were negative in the multiplex assay. Taken together, our results indicate that the one-step multiplex RT-PCR developed here for the detection of GAR, GBR, GCR, BCV, and BToV can be expected to be a useful tool for the rapid and cost-effective diagnosis and surveillance of viral diarrhea in adult cattle.

Affiliations:

Links toward previous steps (curation, corpus...)

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Le document en format XML

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<series><title level="j" type="main">Archives of virology</title>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Bovine</term>
<term>Detection</term>
<term>Diarrhea</term>
<term>Multiplex polymerase chain reaction</term>
<term>Reverse transcription polymerase chain reaction</term>
<term>Veterinary</term>
</keywords>
<keywords scheme="Pascal" xml:lang="fr"><term>Bovin</term>
<term>Réaction chaîne polymérase multiplex</term>
<term>Réaction chaîne polymérase RT</term>
<term>Détection</term>
<term>Diarrhée</term>
<term>Vétérinaire</term>
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</teiHeader>
<front><div type="abstract" xml:lang="en">A one-step multiplex reverse transcription (RT)-PCR method was developed for the simultaneous detection of five viruses causing diarrhea in adult cattle: bovine group A rotavirus (GAR), bovine group B rotavirus (GBR), bovine group C rotavirus (GCR), bovine corona-virus (BCV), and bovine torovirus (BToV). The detection limit of the one-step multiplex RT-PCR for GAR, GCR, BCV, and BToV was 10<sup>2</sup>
, 10°, 10<sup>1</sup>
, and 10<sup>2</sup>
 TCID<sub>50</sub>
/ml, respectively, and that for GBR was 10<sup>6</sup>
 copies/ml. The one-step multiplex RT-PCR with newly designed primers to detect GAR had higher sensitivity than a single RT-PCR with conventional primers, with no false-positive reactions observed for ten other kinds of bovine RNA viruses To assess its field applicability, 59 of 60 fecal samples containing one of these five viruses from all 25 epidemic diarrhea outbreaks in adult cattle were positive in the one-step multiplex RT-PCR assay. Furthermore, using four additional fecal samples containing two viruses (GBR and BCV or BToV), two amplified products of the expected sizes were obtained simultaneously. In contrast, all 80 fecal samples lacking the five target viruses from normal adult cattle were negative in the multiplex assay. Taken together, our results indicate that the one-step multiplex RT-PCR developed here for the detection of GAR, GBR, GCR, BCV, and BToV can be expected to be a useful tool for the rapid and cost-effective diagnosis and surveillance of viral diarrhea in adult cattle.</div>
</front>
</TEI>
<affiliations><list><country><li>Japon</li>
</country>
<region><li>Préfecture de Gifu</li>
<li>Région du Chūbu</li>
</region>
<settlement><li>Gifu (ville)</li>
</settlement>
<orgName><li>Université de Gifu</li>
</orgName>
</list>
<tree><country name="Japon"><noRegion><name sortKey="Fukuda, Masaharu" sort="Fukuda, Masaharu" uniqKey="Fukuda M" first="Masaharu" last="Fukuda">Masaharu Fukuda</name>
</noRegion>
<name sortKey="Aita, Tsunehiko" sort="Aita, Tsunehiko" uniqKey="Aita T" first="Tsunehiko" last="Aita">Tsunehiko Aita</name>
<name sortKey="Fukuda, Masaharu" sort="Fukuda, Masaharu" uniqKey="Fukuda M" first="Masaharu" last="Fukuda">Masaharu Fukuda</name>
<name sortKey="Kuga, Kazufumi" sort="Kuga, Kazufumi" uniqKey="Kuga K" first="Kazufumi" last="Kuga">Kazufumi Kuga</name>
<name sortKey="Kuga, Kazufumi" sort="Kuga, Kazufumi" uniqKey="Kuga K" first="Kazufumi" last="Kuga">Kazufumi Kuga</name>
<name sortKey="Mase, Masaji" sort="Mase, Masaji" uniqKey="Mase M" first="Masaji" last="Mase">Masaji Mase</name>
<name sortKey="Mase, Masaji" sort="Mase, Masaji" uniqKey="Mase M" first="Masaji" last="Mase">Masaji Mase</name>
<name sortKey="Miyazaki, Ayako" sort="Miyazaki, Ayako" uniqKey="Miyazaki A" first="Ayako" last="Miyazaki">Ayako Miyazaki</name>
<name sortKey="Sugiyama, Makoto" sort="Sugiyama, Makoto" uniqKey="Sugiyama M" first="Makoto" last="Sugiyama">Makoto Sugiyama</name>
<name sortKey="Suzuki, Tohru" sort="Suzuki, Tohru" uniqKey="Suzuki T" first="Tohru" last="Suzuki">Tohru Suzuki</name>
<name sortKey="Tasei, Keito" sort="Tasei, Keito" uniqKey="Tasei K" first="Keito" last="Tasei">Keito Tasei</name>
<name sortKey="Tsunemitsu, Hiroshi" sort="Tsunemitsu, Hiroshi" uniqKey="Tsunemitsu H" first="Hiroshi" last="Tsunemitsu">Hiroshi Tsunemitsu</name>
<name sortKey="Tsunemitsu, Hiroshi" sort="Tsunemitsu, Hiroshi" uniqKey="Tsunemitsu H" first="Hiroshi" last="Tsunemitsu">Hiroshi Tsunemitsu</name>
</country>
</tree>
</affiliations>
</record>

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Development and application of one-step multiplex reverse transcription PCR for simultaneous detection of five diarrheal viruses in adult cattle

Development and application of one-step multiplex reverse transcription PCR for simultaneous detection of five diarrheal viruses in adult cattle

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