One-Pot Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP) for Detecting MERS-CoV
Identifieur interne : 000C53 ( Main/Exploration ); précédent : 000C52; suivant : 000C54One-Pot Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP) for Detecting MERS-CoV
Auteurs : Se Hee Lee [Corée du Sud] ; Yun Hee Baek [Corée du Sud] ; Yang-Hoon Kim [Corée du Sud] ; Young-Ki Choi [Corée du Sud] ; Min-Suk Song [Corée du Sud] ; Ji-Young Ahn [Corée du Sud]Source :
- Frontiers in Microbiology [ 1664-302X ] ; 2017.
Abstract
Due to the limitation of rapid development of specific antiviral drug or vaccine for novel emerging viruses, an accurate and rapid diagnosis is a key to manage the virus spread. We developed an efficient and rapid method with high specificity for the Middle East Respiratory Syndrome coronavirus (MERS-CoV), based on one-pot reverse transcription loop-mediated isothermal amplification (one-pot RT-LAMP). A set of six LAMP primers [F3, B3, FIP, BIP, LF (Loop-F), and LB (Loop-B)] were designed using the sequence of nucleocapsid (N) gene with optimized RT-LAMP enzyme conditions: 100 U M-MLV RTase and 4 U
Url:
DOI: 10.3389/fmicb.2016.02166
PubMed: 28119682
PubMed Central: 5220095
Affiliations:
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<front><div type="abstract" xml:lang="en"><p>Due to the limitation of rapid development of specific antiviral drug or vaccine for novel emerging viruses, an accurate and rapid diagnosis is a key to manage the virus spread. We developed an efficient and rapid method with high specificity for the Middle East Respiratory Syndrome coronavirus (MERS-CoV), based on one-pot reverse transcription loop-mediated isothermal amplification (one-pot RT-LAMP). A set of six LAMP primers [F3, B3, FIP, BIP, LF (Loop-F), and LB (Loop-B)] were designed using the sequence of nucleocapsid (N) gene with optimized RT-LAMP enzyme conditions: 100 U M-MLV RTase and 4 U <italic>Bst</italic>
polymerase, implying that the reaction was able to detect four infectious viral genome copies of MERS-CoV within a 60 min reaction time period. Significantly, EvaGreen dye has better signal read-out properties in one-pot RT-LAMP reaction and is more compatible with DNA polymerase than SYBR green I. Isothermally amplified specific N genes were further evaluated using field-deployable microchamber devices, leading to the specific identification of as few as 0.4 infectious viral genome copies, with no cross-reaction to the other acute respiratory disease viruses, including influenza type A (H1N1 and H3N2), type B, human coronavirus 229E, and human metapneumovirus. This sensitive, specific and feasible method provides a large-scale technical support in emergencies, and is also applied as a sample-to-detection module in Point of Care Testing devices.</p>
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</TEI>
<affiliations><list><country><li>Corée du Sud</li>
</country>
</list>
<tree><country name="Corée du Sud"><noRegion><name sortKey="Lee, Se Hee" sort="Lee, Se Hee" uniqKey="Lee S" first="Se Hee" last="Lee">Se Hee Lee</name>
</noRegion>
<name sortKey="Ahn, Ji Young" sort="Ahn, Ji Young" uniqKey="Ahn J" first="Ji-Young" last="Ahn">Ji-Young Ahn</name>
<name sortKey="Baek, Yun Hee" sort="Baek, Yun Hee" uniqKey="Baek Y" first="Yun Hee" last="Baek">Yun Hee Baek</name>
<name sortKey="Choi, Young Ki" sort="Choi, Young Ki" uniqKey="Choi Y" first="Young-Ki" last="Choi">Young-Ki Choi</name>
<name sortKey="Kim, Yang Hoon" sort="Kim, Yang Hoon" uniqKey="Kim Y" first="Yang-Hoon" last="Kim">Yang-Hoon Kim</name>
<name sortKey="Song, Min Suk" sort="Song, Min Suk" uniqKey="Song M" first="Min-Suk" last="Song">Min-Suk Song</name>
</country>
</tree>
</affiliations>
</record>
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