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DC FieldValueLanguage
dc.contributorFaculty of Health and Social Sciencesen_US
dc.creatorYuen, Suk-han-
dc.identifier.urihttps://theses.lib.polyu.edu.hk/handle/200/5688-
dc.languageEnglishen_US
dc.publisherHong Kong Polytechnic University-
dc.rightsAll rights reserveden_US
dc.titleMultiplex real-time reverse transcription PCR for detection of influenza virusesen_US
dcterms.abstractInfluenza viral infection is a global concern due to its high morbidity and mortality rate. Anti-viral drugs for influenza virus infections should be initiated within 48 hours of onset symptoms. Viral culture is time-consuming for diagnostic purposes and a method that will rapidly detect viruses is essential for management of patients with upper respiratory tract infections. Reverse transcription polymerase chain reaction (RT-PCR) is a useful tool for detection of influenza viruses. This study developed a two-step multiplex RT-PCR assay for detection of influenza A. Species of influenza A and its subtypes (H1, H3, H5, and H1 human swine) were identified using SYTO9 and Roche LightCycler 480. Five sets of primer pairs (M gene, H1, H3, H5, and H1 human swine) were designed for these strains and were optimized. Each primer pair generated a unique amplicon, evaluated by melting temperature due to different sequences of nucleotide bases. Due to non-specific signals found in melting curve analysis, two sets of primer pairs (M gene and H3) failed to be optimized. The duplex assay for seasonal H1 influenza A & human swine influenza (HSI) was evaluated using 86 nasopharyngeal aspirate samples, collected from patients with respiratory viruses in 2009. The seasonal H1 and HSI-specific primer sets had a detection limit approximately equivalent to 2.47 x 10⁹ and 2.18 x 10⁹ genome copies/ul, respectively. Sensitivity and specificity of duplex seasonal H1 and HSI specific real-time RT-PCR were 62% and 98%, respectively.en_US
dcterms.extentxi, 78 leaves : ill. ; 30 cm.en_US
dcterms.isPartOfPolyU Electronic Thesesen_US
dcterms.issued2010en_US
dcterms.educationalLevelAll Masteren_US
dcterms.educationalLevelM.Sc.en_US
dcterms.LCSHHong Kong Polytechnic University -- Dissertationsen_US
dcterms.LCSHInfluenza viruses -- China -- Hong Kongen_US
dcterms.LCSHPolymerase chain reaction -- Diagnostic useen_US
dcterms.accessRightsrestricted accessen_US

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Please use this identifier to cite or link to this item: https://theses.lib.polyu.edu.hk/handle/200/5688