Differential detection of turkey coronavirus, infectious bronchitis virus, and bovine coronavirus by a multiplex polymerase chain reaction.
Identifieur interne : 003F22 ( Main/Exploration ); précédent : 003F21; suivant : 003F23Differential detection of turkey coronavirus, infectious bronchitis virus, and bovine coronavirus by a multiplex polymerase chain reaction.
Auteurs : C C Loa [États-Unis] ; T L Lin ; C C Wu ; T A Bryan ; T A Hooper ; D L SchraderSource :
- Journal of virological methods [ 0166-0934 ] ; 2006.
Descripteurs français
- KwdFr :
- ARN viral (génétique), Amorces ADN, Animaux, Coronavirus bovin (), Coronavirus bovin (génétique), Coronavirus de la dinde (), Coronavirus de la dinde (génétique), Dindons, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (génétique), Gènes viraux, Infections à coronavirus (diagnostic), Protéines de l'enveloppe virale (génétique), Protéines nucléocapside (génétique), Réaction de polymérisation en chaîne (), Sensibilité et spécificité, Spécificité d'espèce, Virus de la bronchite infectieuse (), Virus de la bronchite infectieuse (génétique).
- MESH :
- diagnostic : Infections à coronavirus.
- génétique : ARN viral, Coronavirus bovin, Coronavirus de la dinde, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines nucléocapside, Virus de la bronchite infectieuse.
- Amorces ADN, Animaux, Coronavirus bovin, Coronavirus de la dinde, Dindons, Glycoprotéine de spicule des coronavirus, Gènes viraux, Réaction de polymérisation en chaîne, Sensibilité et spécificité, Spécificité d'espèce, Virus de la bronchite infectieuse.
English descriptors
- KwdEn :
- Animals, Coronavirus Infections (diagnosis), Coronavirus, Bovine (chemistry), Coronavirus, Bovine (genetics), Coronavirus, Turkey (chemistry), Coronavirus, Turkey (genetics), DNA Primers, Genes, Viral, Infectious bronchitis virus (chemistry), Infectious bronchitis virus (genetics), Membrane Glycoproteins (genetics), Nucleocapsid Proteins (genetics), Polymerase Chain Reaction (methods), RNA, Viral (genetics), Sensitivity and Specificity, Species Specificity, Spike Glycoprotein, Coronavirus, Turkeys, Viral Envelope Proteins (genetics).
- MESH :
- chemical , genetics : Membrane Glycoproteins, Nucleocapsid Proteins, RNA, Viral, Viral Envelope Proteins.
- chemical : DNA Primers, Spike Glycoprotein, Coronavirus.
- chemistry : Coronavirus, Bovine, Coronavirus, Turkey, Infectious bronchitis virus.
- diagnosis : Coronavirus Infections.
- genetics : Coronavirus, Bovine, Coronavirus, Turkey, Infectious bronchitis virus.
- methods : Polymerase Chain Reaction.
- Animals, Genes, Viral, Sensitivity and Specificity, Species Specificity, Turkeys.
Abstract
The objective of the present study was to develop a multiplex polymerase chain reaction (PCR) method for differential detection of turkey coronavirus (TCoV), infectious bronchitis coronavirus (IBV), and bovine coronavirus (BCoV). Primers were designed from conserved or variable regions of nucleocapsid (N) or spike (S) protein gene among TCoV, IBV, and BCoV and used in the same PCR reaction. Reverse transcription followed by the PCR reaction was used to amplify a portion of N or S gene of the corresponding coronaviruses. The PCR products were detected on agarose gel stained with ethidium bromide. Two PCR products, a 356-bp band corresponding to N gene and a 727-bp band corresponding to S gene, were obtained for TCoV isolates. In contrast, one PCR product of 356 bp corresponding to a fragment of N gene was obtained for IBV strains and one PCR product of 568 bp corresponding to a fragment of S gene was obtained for BCoV. There were no PCR products with the same primers for Newcastle disease virus, Marek's disease virus, turkey pox virus, pigeon pox virus, fowl pox virus, reovirus, infectious bursal disease virus, enterovirus, astrovirus, Salmonella enterica, Escherichia coli, and Mycoplasma gallisepticum. Performance of the assay with serially diluted RNA demonstrated that the multiplex PCR could detect 4.8x10(-3) microg of TCoV RNA, 4.6x10(-4) microg of IBV RNA, and 8.0x10(-2) microg of BCoV RNA. These results indicated that the multiplex PCR as established in the present study is a rapid, sensitive, and specific method for differential detection of TCoV, IBV, and BCoV in a single PCR reaction.
DOI: 10.1016/j.jviromet.2005.07.007
PubMed: 16137773
Affiliations:
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Le document en format XML
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xml:lang="fr"><term>ARN viral (génétique)</term><term>Amorces ADN</term><term>Animaux</term><term>Coronavirus bovin ()</term><term>Coronavirus bovin (génétique)</term><term>Coronavirus de la dinde ()</term><term>Coronavirus de la dinde (génétique)</term><term>Dindons</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires (génétique)</term><term>Gènes viraux</term><term>Infections à coronavirus (diagnostic)</term><term>Protéines de l'enveloppe virale (génétique)</term><term>Protéines nucléocapside (génétique)</term><term>Réaction de polymérisation en chaîne ()</term><term>Sensibilité et spécificité</term><term>Spécificité d'espèce</term><term>Virus de la bronchite infectieuse ()</term><term>Virus de la bronchite infectieuse (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Membrane Glycoproteins</term><term>Nucleocapsid Proteins</term><term>RNA, Viral</term><term>Viral Envelope 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detection of turkey coronavirus (TCoV), infectious bronchitis coronavirus (IBV), and bovine coronavirus (BCoV). Primers were designed from conserved or variable regions of nucleocapsid (N) or spike (S) protein gene among TCoV, IBV, and BCoV and used in the same PCR reaction. Reverse transcription followed by the PCR reaction was used to amplify a portion of N or S gene of the corresponding coronaviruses. The PCR products were detected on agarose gel stained with ethidium bromide. Two PCR products, a 356-bp band corresponding to N gene and a 727-bp band corresponding to S gene, were obtained for TCoV isolates. In contrast, one PCR product of 356 bp corresponding to a fragment of N gene was obtained for IBV strains and one PCR product of 568 bp corresponding to a fragment of S gene was obtained for BCoV. There were no PCR products with the same primers for Newcastle disease virus, Marek's disease virus, turkey pox virus, pigeon pox virus, fowl pox virus, reovirus, infectious bursal disease virus, enterovirus, astrovirus, Salmonella enterica, Escherichia coli, and Mycoplasma gallisepticum. Performance of the assay with serially diluted RNA demonstrated that the multiplex PCR could detect 4.8x10(-3) microg of TCoV RNA, 4.6x10(-4) microg of IBV RNA, and 8.0x10(-2) microg of BCoV RNA. These results indicated that the multiplex PCR as established in the present study is a rapid, sensitive, and specific method for differential detection of TCoV, IBV, and BCoV in a single PCR reaction.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Indiana</li></region></list><tree><noCountry><name sortKey="Bryan, T A" sort="Bryan, T A" uniqKey="Bryan T" first="T A" last="Bryan">T A Bryan</name><name sortKey="Hooper, T A" sort="Hooper, T A" uniqKey="Hooper T" first="T A" last="Hooper">T A Hooper</name><name sortKey="Lin, T L" sort="Lin, T L" uniqKey="Lin T" first="T L" last="Lin">T L Lin</name><name sortKey="Schrader, D L" sort="Schrader, D L" uniqKey="Schrader D" first="D L" last="Schrader">D L Schrader</name><name sortKey="Wu, C C" sort="Wu, C C" uniqKey="Wu C" first="C C" last="Wu">C C Wu</name></noCountry><country name="États-Unis"><region name="Indiana"><name sortKey="Loa, C C" sort="Loa, C C" uniqKey="Loa C" first="C C" last="Loa">C C Loa</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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