M‐specific reverse transcription loop‐mediated isothermal amplification for detection of pandemic (H1N1) 2009 virus
Identifieur interne : 000D28 ( Main/Exploration ); précédent : 000D27; suivant : 000D29M‐specific reverse transcription loop‐mediated isothermal amplification for detection of pandemic (H1N1) 2009 virus
Auteurs : Min-Shiuh Lee ; Hung-Chang Shih ; Jang-Jih Lu ; Mei-Chi Su ; Ming-Chung Deng ; Chia-Chen Wu ; Fong-Yuan Lin ; Kuan-Hsun Lin ; Po-Yen Chen [Taïwan] ; Wei-Li HsuSource :
- European Journal of Clinical Investigation [ 0014-2972 ] ; 2011-04.
English descriptors
- Teeft :
- Analytical sensitivity, Animal health research institute, Antigen detection tests, Assay, Avian subtypes, Clin, Clin microbiol, Clin virol, Clinic nasopharyngeal swabs, Clinic samples, Clinical investigation, Clinical investigation journal foundation detection, Clinical nasopharyngeal swabs, Clinical samples, Consensus sequence, Copy number, Cultured pandemic, Eiken chemical, European journal, Gene sequences, Genetic stability, Graduate institute, Human echovirus, Isothermal, Isothermal assay, Kuang road, Laboratory medicine, Lamp assay, Limited equipment, Major drawback, Matrix protein, Mutation trend analysis, Nasopharyngeal, Nasopharyngeal swabs, National chung hsing university, National university, Negative control, Novel pandemic, Other viruses, Pandemic, Pandemic spanish, Present study, Primer, Public health, Rapid antigen test, Rapid detection, Reference strain, Sequence alignment, Serial dilutions, Stichting european society, Subtypes, Swab, Taichung, Taichung veterans, Thermocycling conditions, Transcription, University hospital, Veterinary medicine, Viral, Viral isolation, Virol, Virol methods, Virus, Virus isolation, Virus samples, Virus strains.
Abstract
Eur J Clin Invest 2011; 41 (4): 434–441 Abstract: Background Since the initial outbreak in March 2009, the novel pandemic (H1N1) 2009 virus has affected individuals worldwide and caused over 18 138 deaths. There is an urgent need for the development of an easy, accurate and simple method for the diagnosis of this novel pandemic virus. Design Reverse transcription loop‐mediated isothermal amplification assay (RT‐LAMP) with primers targeting the M segment was established for the rapid differential diagnosis of pandemic (H1N1) 2009 virus. The performance of this assay was characterized using 111 clinic nasopharyngeal swabs, and the diagnosis accuracy was compared with real‐time reverse transcription PCR (RRT‐PCR) and virus isolation, the latter being the reference standard. Results This method successfully detected pandemic (H1N1) 2009 virus with a detection limit of approximately 20 copies of the target RNA per reaction, which is a comparably sensitivity to the RRT‐PCR assay. Furthermore, this assay was able to discriminate pandemic (H1N1) 2009 virus from seasonal influenza viruses, such as H1N1 and H3N2, and other respiratory viruses (parainfluenza type 2 and 3, adenovirus, echovirus 7, and coxsackievirus A10). Based on validation by virus isolation, the specificity and sensitivity of this M‐specific RT‐LAMP assay were 100% and 98·25%, respectively. Moreover, the RT‐LAMP amplification of most positive samples (46 out of 56) was achieved in < 20 min. Conclusions This is an accurate and fast analysis system suitable for general diagnostic laboratories with only limited equipment, e.g. first‐line health care centre. This assay will help clinicians and public health officials to react effectively during an outbreak.
Url:
DOI: 10.1111/j.1365-2362.2010.02427.x
Affiliations:
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Shih</name><affiliation><wicri:noCountry code="subField">Hospital</wicri:noCountry></affiliation></author><author><name sortKey="Lu, Jang Ih" sort="Lu, Jang Ih" uniqKey="Lu J" first="Jang-Jih" last="Lu">Jang-Jih Lu</name><affiliation><wicri:noCountry code="subField">Hospital</wicri:noCountry></affiliation></author><author><name sortKey="Su, Mei Hi" sort="Su, Mei Hi" uniqKey="Su M" first="Mei-Chi" last="Su">Mei-Chi Su</name><affiliation><wicri:noCountry code="subField">Hospital</wicri:noCountry></affiliation></author><author><name sortKey="Deng, Ming Hung" sort="Deng, Ming Hung" uniqKey="Deng M" first="Ming-Chung" last="Deng">Ming-Chung Deng</name><affiliation><wicri:noCountry code="subField">Taipei</wicri:noCountry></affiliation></author><author><name sortKey="Wu, Chia Hen" sort="Wu, Chia Hen" uniqKey="Wu C" first="Chia-Chen" last="Wu">Chia-Chen Wu</name><affiliation><wicri:noCountry code="subField">University</wicri:noCountry></affiliation></author><author><name sortKey="Lin, Fong Uan" sort="Lin, Fong Uan" uniqKey="Lin F" first="Fong-Yuan" last="Lin">Fong-Yuan Lin</name><affiliation><wicri:noCountry code="subField">University</wicri:noCountry></affiliation></author><author><name sortKey="Lin, Kuan Sun" sort="Lin, Kuan Sun" uniqKey="Lin K" first="Kuan-Hsun" last="Lin">Kuan-Hsun Lin</name><affiliation><wicri:noCountry code="subField">University</wicri:noCountry></affiliation></author><author><name sortKey="Chen, Po En" sort="Chen, Po En" uniqKey="Chen P" first="Po-Yen" last="Chen">Po-Yen Chen</name><affiliation wicri:level="1"><country xml:lang="fr">Taïwan</country><wicri:regionArea>Department of Pediatrics, Taichung Veterans General Hospital, Taichung</wicri:regionArea><wicri:noRegion>Taichung</wicri:noRegion></affiliation></author><author><name sortKey="Hsu, Wei I" sort="Hsu, Wei I" uniqKey="Hsu W" first="Wei-Li" last="Hsu">Wei-Li Hsu</name><affiliation><wicri:noCountry code="subField">University</wicri:noCountry></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">European Journal of Clinical Investigation</title><title level="j" type="alt">EUROPEAN JOURNAL OF CLINICAL INVESTIGATION</title><idno type="ISSN">0014-2972</idno><idno type="eISSN">1365-2362</idno><imprint><biblScope unit="vol">41</biblScope><biblScope unit="issue">4</biblScope><biblScope unit="page" from="434">434</biblScope><biblScope unit="page" to="441">441</biblScope><biblScope unit="page-count">8</biblScope><publisher>Blackwell Publishing Ltd</publisher><pubPlace>Oxford, UK</pubPlace><date type="published" when="2011-04">2011-04</date></imprint><idno type="ISSN">0014-2972</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0014-2972</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Analytical sensitivity</term><term>Animal health research institute</term><term>Antigen detection tests</term><term>Assay</term><term>Avian subtypes</term><term>Clin</term><term>Clin microbiol</term><term>Clin virol</term><term>Clinic nasopharyngeal swabs</term><term>Clinic samples</term><term>Clinical investigation</term><term>Clinical investigation journal foundation detection</term><term>Clinical nasopharyngeal swabs</term><term>Clinical samples</term><term>Consensus sequence</term><term>Copy number</term><term>Cultured pandemic</term><term>Eiken chemical</term><term>European journal</term><term>Gene sequences</term><term>Genetic stability</term><term>Graduate institute</term><term>Human echovirus</term><term>Isothermal</term><term>Isothermal assay</term><term>Kuang road</term><term>Laboratory medicine</term><term>Lamp assay</term><term>Limited equipment</term><term>Major drawback</term><term>Matrix protein</term><term>Mutation trend analysis</term><term>Nasopharyngeal</term><term>Nasopharyngeal swabs</term><term>National chung hsing university</term><term>National university</term><term>Negative control</term><term>Novel pandemic</term><term>Other viruses</term><term>Pandemic</term><term>Pandemic spanish</term><term>Present study</term><term>Primer</term><term>Public health</term><term>Rapid antigen test</term><term>Rapid detection</term><term>Reference strain</term><term>Sequence alignment</term><term>Serial dilutions</term><term>Stichting european society</term><term>Subtypes</term><term>Swab</term><term>Taichung</term><term>Taichung veterans</term><term>Thermocycling conditions</term><term>Transcription</term><term>University hospital</term><term>Veterinary medicine</term><term>Viral</term><term>Viral isolation</term><term>Virol</term><term>Virol methods</term><term>Virus</term><term>Virus isolation</term><term>Virus samples</term><term>Virus strains</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Eur J Clin Invest 2011; 41 (4): 434–441 Abstract: Background Since the initial outbreak in March 2009, the novel pandemic (H1N1) 2009 virus has affected individuals worldwide and caused over 18 138 deaths. There is an urgent need for the development of an easy, accurate and simple method for the diagnosis of this novel pandemic virus. Design Reverse transcription loop‐mediated isothermal amplification assay (RT‐LAMP) with primers targeting the M segment was established for the rapid differential diagnosis of pandemic (H1N1) 2009 virus. The performance of this assay was characterized using 111 clinic nasopharyngeal swabs, and the diagnosis accuracy was compared with real‐time reverse transcription PCR (RRT‐PCR) and virus isolation, the latter being the reference standard. Results This method successfully detected pandemic (H1N1) 2009 virus with a detection limit of approximately 20 copies of the target RNA per reaction, which is a comparably sensitivity to the RRT‐PCR assay. Furthermore, this assay was able to discriminate pandemic (H1N1) 2009 virus from seasonal influenza viruses, such as H1N1 and H3N2, and other respiratory viruses (parainfluenza type 2 and 3, adenovirus, echovirus 7, and coxsackievirus A10). Based on validation by virus isolation, the specificity and sensitivity of this M‐specific RT‐LAMP assay were 100% and 98·25%, respectively. Moreover, the RT‐LAMP amplification of most positive samples (46 out of 56) was achieved in < 20 min. Conclusions This is an accurate and fast analysis system suitable for general diagnostic laboratories with only limited equipment, e.g. first‐line health care centre. This assay will help clinicians and public health officials to react effectively during an outbreak.</div></front></TEI><affiliations><list><country><li>Taïwan</li></country></list><tree><noCountry><name sortKey="Deng, Ming Hung" sort="Deng, Ming Hung" uniqKey="Deng M" first="Ming-Chung" last="Deng">Ming-Chung Deng</name><name sortKey="Hsu, Wei I" sort="Hsu, Wei I" uniqKey="Hsu W" first="Wei-Li" last="Hsu">Wei-Li Hsu</name><name sortKey="Lee, Min Hiuh" sort="Lee, Min Hiuh" uniqKey="Lee M" first="Min-Shiuh" last="Lee">Min-Shiuh Lee</name><name sortKey="Lin, Fong Uan" sort="Lin, Fong Uan" uniqKey="Lin F" first="Fong-Yuan" last="Lin">Fong-Yuan Lin</name><name sortKey="Lin, Kuan Sun" sort="Lin, Kuan Sun" uniqKey="Lin K" first="Kuan-Hsun" last="Lin">Kuan-Hsun Lin</name><name sortKey="Lu, Jang Ih" sort="Lu, Jang Ih" uniqKey="Lu J" first="Jang-Jih" last="Lu">Jang-Jih Lu</name><name sortKey="Shih, Hung Hang" sort="Shih, Hung Hang" uniqKey="Shih H" first="Hung-Chang" last="Shih">Hung-Chang Shih</name><name sortKey="Su, Mei Hi" sort="Su, Mei Hi" uniqKey="Su M" first="Mei-Chi" last="Su">Mei-Chi Su</name><name sortKey="Wu, Chia Hen" sort="Wu, Chia Hen" uniqKey="Wu C" first="Chia-Chen" last="Wu">Chia-Chen Wu</name></noCountry><country name="Taïwan"><noRegion><name sortKey="Chen, Po En" sort="Chen, Po En" uniqKey="Chen P" first="Po-Yen" last="Chen">Po-Yen Chen</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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