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developed and compared with rapid antigen enzyme immunoassay (AgEIA) and enhanced virus culture for rapid detection of influenza A viruses in stored and prospectively collected respiratory specimens. Specific hybridization probes were used for simultaneous detection and differentiation between H1N1 and H3N2 subtypes. The sensitivity of the RT-PCR for influenza A H1N1 was 120 copies and H3N2 350 copies of in vitro transcribed RNA. A specimen was considered positive for influenza A when it was culture positive or at least two methods yielded a positive test result. Using these criteria, with stored samples, the RT-PCR sensitivity, specificity, positive and negative predictive values were 82.9, 95.5, 98.9 and 52.5%, respectively. In specimens collected prospectively the RT-PCR sensitivity, specificity, positive and negative predictive values were 100, 87.9, 82.8 and 100%, respectively. There was complete concordance with subtype differentiation by hybridization probe melting temperature analysis and haemagglutination inhibition assay.","publication_date":"2004,,","publication_name":"Journal of Virological Methods","grobid_abstract_attachment_id":"43214109"},"document_type":"paper","pre_hit_view_count_baseline":null,"quality":"high","language":"en","title":"Rapid detection and simultaneous subtype differentiation of influenza A viruses by real time PCR","broadcastable":true,"draft":null,"has_indexable_attachment":true,"indexable":true}}["work"]; window.loswp.workCoauthors = [44074767]; window.loswp.locale = "en"; window.loswp.countryCode = "SG"; window.loswp.cwvAbTestBucket = ""; window.loswp.designVariant = "ds_vanilla"; window.loswp.fullPageMobileSutdModalVariant = "control"; window.loswp.useOptimizedScribd4genScript = false; window.loswp.appleClientId = 'edu.academia.applesignon';</script><script defer="" src="https://accounts.google.com/gsi/client"></script><div 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translate="no">download</span><p>Download Free PDF</p></div><div class="ds-work-cover--ribbon-container">Download Free PDF</div><div class="ds-work-cover--ribbon-triangle"></div></button></div></div></div><div class="ds-work-card--work-information"><h1 class="ds-work-card--work-title">Rapid detection and simultaneous subtype differentiation of influenza A viruses by real time PCR</h1><div class="ds-work-card--work-authors ds-work-card--detail"><a class="ds-work-card--author js-wsj-grid-card-author ds2-5-body-md ds2-5-body-link" data-author-id="44074767" href="https://independent.academia.edu/BurrowsJulie"><img alt="Profile image of Julie Burrows" class="ds-work-card--author-avatar" src="//a.academia-assets.com/images/s65_no_pic.png" />Julie Burrows</a></div><div class="ds-work-card--detail"><p class="ds-work-card--detail ds2-5-body-sm">2004, Journal of Virological Methods</p></div><div class="ds-work-card--button-container"><button class="ds2-5-button js-swp-download-button" 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data-landing_url="https://www.academia.edu/22615508/Rapid_detection_and_simultaneous_subtype_differentiation_of_influenza_A_viruses_by_real_time_PCR" data-login_uri="https://www.academia.edu/registrations/google_one_tap" data-moment_callback="onGoogleOneTapEvent" id="g_id_onload"></div><div class="ds-top-related-works--grid-container"><div class="ds-related-content--container ds-top-related-works--container"><h2 class="ds-related-content--heading">Related papers</h2><div class="ds-related-work--container js-wsj-grid-card" data-collection-position="0" data-entity-id="16572978" data-sort-order="default"><a class="ds-related-work--title js-wsj-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/16572978/Comparison_of_three_non_nested_RT_PCR_for_the_detection_of_influenza_A_viruses">Comparison of three non-nested RT-PCR for the detection of influenza A viruses</a><div class="ds-related-work--metadata"><a class="js-wsj-grid-card-author ds2-5-body-sm 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kim</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Journal of microbiology and biotechnology, 2008</p><p class="ds-related-work--abstract ds2-5-body-sm">We developed multiplex RT-PCR assays that can detect and identify 12 hemagglutinin (H1-H12) and 9 neuraminidase (N1-N9) subtypes that are commonly isolated from avian, swine, and human influenza A viruses. 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Furthermore, the multiplex RT-PCR methods efficiently detected mixed infections with at least two different subtypes...</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"Development of multiplex rt-PCR assays for rapid detection and subtyping of influenza type A viruses from clinical specimens","attachmentId":91016025,"attachmentType":"pdf","work_url":"https://www.academia.edu/86601677/Development_of_multiplex_rt_PCR_assays_for_rapid_detection_and_subtyping_of_influenza_type_A_viruses_from_clinical_specimens","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-wsj-grid-card-view-pdf" href="https://www.academia.edu/86601677/Development_of_multiplex_rt_PCR_assays_for_rapid_detection_and_subtyping_of_influenza_type_A_viruses_from_clinical_specimens"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div><div class="ds-related-work--container js-wsj-grid-card" data-collection-position="2" data-entity-id="14063231" data-sort-order="default"><a class="ds-related-work--title js-wsj-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/14063231/Detection_of_Influenza_Virus_Types_A_and_B_and_Type_A_Subtypes_H1_H3_and_H5_by_Multiplex_Polymerase_Chain_Reaction">Detection of Influenza Virus Types A and B and Type A Subtypes (H1, H3, and H5) by Multiplex Polymerase Chain Reaction</a><div class="ds-related-work--metadata"><a class="js-wsj-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="33080744" href="https://independent.academia.edu/ApiradeeTheamboonlers">Apiradee Theamboonlers</a></div><p class="ds-related-work--metadata ds2-5-body-xs">The Tohoku Journal of Experimental Medicine, 2008</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"Detection of Influenza Virus Types A and B and Type A Subtypes (H1, H3, and H5) by Multiplex Polymerase Chain Reaction","attachmentId":44645579,"attachmentType":"pdf","work_url":"https://www.academia.edu/14063231/Detection_of_Influenza_Virus_Types_A_and_B_and_Type_A_Subtypes_H1_H3_and_H5_by_Multiplex_Polymerase_Chain_Reaction","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-wsj-grid-card-view-pdf" href="https://www.academia.edu/14063231/Detection_of_Influenza_Virus_Types_A_and_B_and_Type_A_Subtypes_H1_H3_and_H5_by_Multiplex_Polymerase_Chain_Reaction"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div><div class="ds-related-work--container js-wsj-grid-card" data-collection-position="3" data-entity-id="125566292" data-sort-order="default"><a class="ds-related-work--title js-wsj-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/125566292/Comparison_of_reverse_transcription_PCR_with_tissue_culture_and_other_rapid_diagnostic_assays_for_detection_of_type_A_influenza_virus">Comparison of reverse transcription-PCR with tissue culture and other rapid diagnostic assays for detection of type A influenza virus</a><div class="ds-related-work--metadata"><a class="js-wsj-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="302796858" href="https://independent.academia.edu/EdwardDominguez15">Edward Dominguez</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Journal of Clinical Microbiology, 1996</p><p class="ds-related-work--abstract ds2-5-body-sm">We applied a reverse transcription (RT)-PCR assay for influenza A virus to combined nasal wash-throat swab specimens previously obtained from an outpatient pediatric population with acute respiratory illness during concurrent epidemics of influenza A virus and respiratory syncytial virus. The results of the RT-PCR assay were compared with those previously reported with virus cultivation and commercially available rapid diagnostic kits (E.A. Dominguez, L.H. Taber, and R.B. Couch, J. Clin. Microbiol. 31:2286-2290, 1993). With virus cultivation as the &quot;gold standard&quot;, the RT-PCR assay had a sensitivity, specificity, and efficiency of 95, 98, and 97%, respectively, compared with 75, 100, and 93%, respectively, for the best diagnostic kit (Becton Dickinson Directigen). 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There is need for a sensitive, rapid and cost-effective assay to detect, type and sub-type influenza viruses, as cell culture has a long turnaround time. Nasopharyngeal swabs were collected from patients presenting with influenza-like illness (ILI) at AIIMS OPD and Primary Health Centre Ballabhgarh (Haryana). From June 2007 to January 2009 and then from September to November 2009, of 1567 specimens collected, 544 were randomly selected and were tested by virus culture using Madin-Darby Canine Kidney (MDCK) cells and by reverse transcription polymerase chain reaction (RT-PCR) for influenza A using primers for matrix gene and for influenza B using non-structural gene (NS) primers. All influenza A positives were sub-typed using primers for HA and NA genes of A/H1, A/H3. 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href="https://www.academia.edu/91694383/Comparison_of_multiplex_RT_PCR_with_virus_isolation_for_detection_typing_and_sub_typing_of_influenza_virus_from_influenza_like_illness_cases"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div><div class="ds-related-work--container js-wsj-grid-card" data-collection-position="5" data-entity-id="24922431" data-sort-order="default"><a class="ds-related-work--title js-wsj-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/24922431/Comparison_of_RT_PCR_with_other_diagnostic_assays_for_rapid_detection_of_influenza_viruses">Comparison of RT-PCR with other diagnostic assays for rapid detection of influenza viruses</a><div class="ds-related-work--metadata"><a class="js-wsj-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="48042402" href="https://independent.academia.edu/FabrizioPregliasco">Fabrizio Pregliasco</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Journal of Medical Virology, 1998</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"Comparison of RT-PCR with other diagnostic assays for rapid detection of influenza viruses","attachmentId":45251055,"attachmentType":"pdf","work_url":"https://www.academia.edu/24922431/Comparison_of_RT_PCR_with_other_diagnostic_assays_for_rapid_detection_of_influenza_viruses","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-wsj-grid-card-view-pdf" href="https://www.academia.edu/24922431/Comparison_of_RT_PCR_with_other_diagnostic_assays_for_rapid_detection_of_influenza_viruses"><span 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data-sort-order="default"><a class="ds-related-work--title js-related-work-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/67339712/Simultaneous_Detection_of_Influenza_Viruses_A_and_B_Using_Real_Time_Quantitative_PCR">Simultaneous Detection of Influenza Viruses A and B Using Real-Time Quantitative PCR</a><div class="ds-related-work--metadata"><a class="js-related-work-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="34181292" href="https://umcu.academia.edu/MoniqueNijhuis">Monique Nijhuis</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Journal of Clinical Microbiology, 2001</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"Simultaneous Detection of Influenza Viruses A and B Using Real-Time Quantitative 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detection tests for distinguishing between influenza virus type A and B among children less than 5 years, using direct immunofluorescence assay and RT-PCR techniques</a><div class="ds-related-work--metadata"><a class="js-related-work-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="247943411" href="https://independent.academia.edu/hassannaif2">hassan naif</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Journal of Biology Agriculture and Healthcare, 2015</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"Rapid detection tests for distinguishing between influenza virus type A and B among children less than 5 years, using direct immunofluorescence assay and RT-PCR 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class="ds-related-work--metadata ds2-5-body-xs">2005</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"Evaluation of a Multiplex Real-Time Reverse Transcriptase PCR Assay for Detection and Differentiation of Influenza Viruses A and B during the 2001-2002 Influenza Season in Israel","attachmentId":31509497,"attachmentType":"pdf","work_url":"https://www.academia.edu/3882248/Evaluation_of_a_Multiplex_Real_Time_Reverse_Transcriptase_PCR_Assay_for_Detection_and_Differentiation_of_Influenza_Viruses_A_and_B_during_the_2001_2002_Influenza_Season_in_Israel","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-related-work-grid-card-view-pdf" href="https://www.academia.edu/3882248/Evaluation_of_a_Multiplex_Real_Time_Reverse_Transcriptase_PCR_Assay_for_Detection_and_Differentiation_of_Influenza_Viruses_A_and_B_during_the_2001_2002_Influenza_Season_in_Israel"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div><div class="ds-related-work--container js-related-work-sidebar-card" data-collection-position="12" data-entity-id="104460015" data-sort-order="default"><a class="ds-related-work--title js-related-work-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/104460015/Influenza_diagnosis_From_dark_isolation_into_the_molecular_light">Influenza diagnosis: From dark isolation into the molecular light</a><div class="ds-related-work--metadata"><a class="js-related-work-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="145936398" href="https://independent.academia.edu/Jamesdouglas65">James douglas</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Journal of Infection, 1999</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"Influenza diagnosis: From dark isolation into the molecular light","attachmentId":104183865,"attachmentType":"pdf","work_url":"https://www.academia.edu/104460015/Influenza_diagnosis_From_dark_isolation_into_the_molecular_light","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-related-work-grid-card-view-pdf" href="https://www.academia.edu/104460015/Influenza_diagnosis_From_dark_isolation_into_the_molecular_light"><span 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