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PCR detection of St...
PCR detection of Streptococcus pneumoniae and Haemophilus influenzae in pneumonia patients
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- Abdeldaim, Guma M. K. (författare)
- Uppsala universitet,Institutionen för medicinska vetenskaper,Jonas Blomberg
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- Herrmann, Björn, Associate professor (preses)
- Uppsala universitet,Klinisk bakteriologi
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- Blomberg, Jonas, professor (preses)
- Uppsala universitet,Klinisk virologi
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- Strålin, Kristoffer, MD, PhD (preses)
- Department of infectious diseases, Örebro university hospital
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- Henriques, Birgitta, Professor (opponent)
- Swedish Institute for Infectious Disease Control, Department of Microbiology
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(creator_code:org_t)
- ISBN 9789155475987
- Uppsala : Acta Universitatis Upsaliensis, 2009
- Engelska 62 s.
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Serie: Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, 1651-6206 ; 479
- Relaterad länk:
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https://uu.diva-port... (primary) (Raw object)
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- PCR is a rapid, reproducible method for nucleic acid detection. However, this technology displays significant deficiencies when applied in clinical microbiology. This work’s aim was to improve current diagnostics and provide sensitive and quantitative real-time PCRs. Paper I describes the development of a sensitive and specific quantitative real-time PCR for the detection of Streptococcus pneumoniae, based on the Spn9802 DNA fragment. Applied to nasopharyngeal aspirates from 166 pneumonia patients, Spn9802 PCR had a sensitivity of 94% and a specificity of 98%. In Paper II the performance of a ply gene PCR for identification of pneumococcal lower respiratory tract infection (LRTI) was evaluated on bronchoalveloar lavage fluids. At the detection limit 103 genome copies/mL, 89% sensitivity but only 43% specificity was achieved. Paper III shows that S. pneumoniae DNA is detectable in plasma from acutely febrile patients. Sensitivities were low (26-42%) for detection of pneumococcal pneumonia, for bacteraemic pneumococcal pneumonia they were 60-70%. Paper IV describes evaluation of four PCR targets for Haemophilus influenzae detection. A real-time PCR based on the P6 gene was developed and applied to 166 CAP patients, using cut-off of 104 genome copies/mL the assay had a sensitivity of 97% and a specificity of 96%. In paper V, the two real-time PCRs presented in papers I and IV were combined with a PCR for detection of Neisseriae meningitidis. The analytical sensitivity of this multiplex real-time PCR was not affected by using a mixture of reagents and a combined DNA standard (S. pneumoniae/H. influenzae) in single tubes. Applied to 156 LRTI patients, this PCR had sensitivities over 90% for S. pneumoniae and H. influenzae, and specificities of 89% and 96%, respectively. In conclusion, real-time PCR assays are useful for the diagnosis of S. pneumoniae and H. influenzae. They enable detection after antibiotic installation, and quantification increases the etiological specificity of pneumonia.
Nyckelord
- Lower respiratory tract infections
- Pneumonia
- Streptococcus pneumoniae
- Haemophilus influenzae
- real-time PCR
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