[Development of Novel Multiplex Real-time RT-PCR Assays for Detection of MERS-CoV Infection].
Identifieur interne : 001144 ( PubMed/Corpus ); précédent : 001143; suivant : 001145[Development of Novel Multiplex Real-time RT-PCR Assays for Detection of MERS-CoV Infection].
Auteurs : Peihua Niu ; Roujian Lu ; Jiaming Lan ; Gaoshan Liu ; Wenling Wang ; Wenjie TanSource :
- Bing du xue bao = Chinese journal of virology [ 1000-8721 ] ; 2016.
English descriptors
- KwdEn :
- China, Coronavirus Infections (diagnosis), Coronavirus Infections (virology), Humans, Middle East Respiratory Syndrome Coronavirus (classification), Middle East Respiratory Syndrome Coronavirus (genetics), Middle East Respiratory Syndrome Coronavirus (isolation & purification), Multiplex Polymerase Chain Reaction (methods), Reverse Transcriptase Polymerase Chain Reaction (methods), Viral Proteins (genetics).
- MESH :
- chemical , genetics : Viral Proteins.
- geographic : China.
- classification : Middle East Respiratory Syndrome Coronavirus.
- diagnosis : Coronavirus Infections.
- genetics : Middle East Respiratory Syndrome Coronavirus.
- isolation & purification : Middle East Respiratory Syndrome Coronavirus.
- methods : Multiplex Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction.
- virology : Coronavirus Infections.
- Humans.
Abstract
We aimed to develop a novel laboratory assay for the detection of Middle East respiratory syndrome coronavirus (MERS-CoV) infection .Several novel multiplex real-time RT-PCR assays were developed using the upE,ORF1 band N2genes of MERS-CoV as targets; the novel assays were compared with previous monoplex real-time RT-PCR assays. For validation, we tested a MERS-CoV strain (hCoVEMC), clinical specimens from patients with fever in Shanghai, and specimens from the first imported MERS case in China. The detection limit of the novel multiplex real-time RT-PCR assays was 10 PFU of MERS-CoV per ml, the same as that in monoplex real-time RT-PCR assays based on upE or N2. The detection was specific for MERS-CoV. In validation using clinical samples, pharyngeal swabs from Shanghai patients were detected as negative, while swabs from the first imported MERS case in China were detected as positive. Using whole blood samples from a MERS case, better detection results were obtained with N2 as the target than upE. We conclude that all the novel assays established in this study could be used for the detection of MERS-CoV; they show potential for improvement compared with monoplex real-time RT-PCR assay based on upE.
PubMed: 29963823
Links to Exploration step
pubmed:29963823Le document en format XML
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<author><name sortKey="Niu, Peihua" sort="Niu, Peihua" uniqKey="Niu P" first="Peihua" last="Niu">Peihua Niu</name>
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<author><name sortKey="Lu, Roujian" sort="Lu, Roujian" uniqKey="Lu R" first="Roujian" last="Lu">Roujian Lu</name>
</author>
<author><name sortKey="Lan, Jiaming" sort="Lan, Jiaming" uniqKey="Lan J" first="Jiaming" last="Lan">Jiaming Lan</name>
</author>
<author><name sortKey="Liu, Gaoshan" sort="Liu, Gaoshan" uniqKey="Liu G" first="Gaoshan" last="Liu">Gaoshan Liu</name>
</author>
<author><name sortKey="Wang, Wenling" sort="Wang, Wenling" uniqKey="Wang W" first="Wenling" last="Wang">Wenling Wang</name>
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<author><name sortKey="Tan, Wenjie" sort="Tan, Wenjie" uniqKey="Tan W" first="Wenjie" last="Tan">Wenjie Tan</name>
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<author><name sortKey="Niu, Peihua" sort="Niu, Peihua" uniqKey="Niu P" first="Peihua" last="Niu">Peihua Niu</name>
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<author><name sortKey="Lu, Roujian" sort="Lu, Roujian" uniqKey="Lu R" first="Roujian" last="Lu">Roujian Lu</name>
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<author><name sortKey="Lan, Jiaming" sort="Lan, Jiaming" uniqKey="Lan J" first="Jiaming" last="Lan">Jiaming Lan</name>
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<author><name sortKey="Liu, Gaoshan" sort="Liu, Gaoshan" uniqKey="Liu G" first="Gaoshan" last="Liu">Gaoshan Liu</name>
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<author><name sortKey="Wang, Wenling" sort="Wang, Wenling" uniqKey="Wang W" first="Wenling" last="Wang">Wenling Wang</name>
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<author><name sortKey="Tan, Wenjie" sort="Tan, Wenjie" uniqKey="Tan W" first="Wenjie" last="Tan">Wenjie Tan</name>
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<series><title level="j">Bing du xue bao = Chinese journal of virology</title>
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<term>Middle East Respiratory Syndrome Coronavirus (classification)</term>
<term>Middle East Respiratory Syndrome Coronavirus (genetics)</term>
<term>Middle East Respiratory Syndrome Coronavirus (isolation & purification)</term>
<term>Multiplex Polymerase Chain Reaction (methods)</term>
<term>Reverse Transcriptase Polymerase Chain Reaction (methods)</term>
<term>Viral Proteins (genetics)</term>
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<keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Middle East Respiratory Syndrome Coronavirus</term>
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<front><div type="abstract" xml:lang="en">We aimed to develop a novel laboratory assay for the detection of Middle East respiratory syndrome coronavirus (MERS-CoV) infection .Several novel multiplex real-time RT-PCR assays were developed using the upE,ORF1 band N2genes of MERS-CoV as targets; the novel assays were compared with previous monoplex real-time RT-PCR assays. For validation, we tested a MERS-CoV strain (hCoVEMC), clinical specimens from patients with fever in Shanghai, and specimens from the first imported MERS case in China. The detection limit of the novel multiplex real-time RT-PCR assays was 10 PFU of MERS-CoV per ml, the same as that in monoplex real-time RT-PCR assays based on upE or N2. The detection was specific for MERS-CoV. In validation using clinical samples, pharyngeal swabs from Shanghai patients were detected as negative, while swabs from the first imported MERS case in China were detected as positive. Using whole blood samples from a MERS case, better detection results were obtained with N2 as the target than upE. We conclude that all the novel assays established in this study could be used for the detection of MERS-CoV; they show potential for improvement compared with monoplex real-time RT-PCR assay based on upE.</div>
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<Abstract><AbstractText>We aimed to develop a novel laboratory assay for the detection of Middle East respiratory syndrome coronavirus (MERS-CoV) infection .Several novel multiplex real-time RT-PCR assays were developed using the upE,ORF1 band N2genes of MERS-CoV as targets; the novel assays were compared with previous monoplex real-time RT-PCR assays. For validation, we tested a MERS-CoV strain (hCoVEMC), clinical specimens from patients with fever in Shanghai, and specimens from the first imported MERS case in China. The detection limit of the novel multiplex real-time RT-PCR assays was 10 PFU of MERS-CoV per ml, the same as that in monoplex real-time RT-PCR assays based on upE or N2. The detection was specific for MERS-CoV. In validation using clinical samples, pharyngeal swabs from Shanghai patients were detected as negative, while swabs from the first imported MERS case in China were detected as positive. Using whole blood samples from a MERS case, better detection results were obtained with N2 as the target than upE. We conclude that all the novel assays established in this study could be used for the detection of MERS-CoV; they show potential for improvement compared with monoplex real-time RT-PCR assay based on upE.</AbstractText>
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