PCR detection of Streptococcus pneumoniae and Haemophilus influenzae in pneumonia patients

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Abdeldaim, Guma M. K.Uppsala universitet,Institutionen för medicinska vetenskaper,Jonas Blomberg (author)

PCR detection of Streptococcus pneumoniae and Haemophilus influenzae in pneumonia patients

BookEnglish2009

Publisher, publication year, extent ...

Uppsala :Acta Universitatis Upsaliensis,2009
62 s.
electronicrdacarrier

Numbers

LIBRIS-ID:oai:DiVA.org:uu-107931
ISBN:9789155475987
https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-107931URI

Supplementary language notes

Language:English
Summary in:English

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SwePubSwePub

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Subject category:vet swepub-contenttype
Subject category:dok swepub-publicationtype

Series

Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine,1651-6206 ;479

Notes

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.

Subject headings and genre

Lower respiratory tract infections
Pneumonia
Streptococcus pneumoniae
Haemophilus influenzae
real-time PCR

Added entries (persons, corporate bodies, meetings, titles ...)

Herrmann, Björn,Associate professorUppsala universitet,Klinisk bakteriologi (thesis advisor)
Blomberg, Jonas,professorUppsala universitet,Klinisk virologi (thesis advisor)
Strålin, Kristoffer,MD, PhDDepartment of infectious diseases, Örebro university hospital (thesis advisor)
Henriques, Birgitta,ProfessorSwedish Institute for Infectious Disease Control, Department of Microbiology (opponent)
Uppsala universitetInstitutionen för medicinska vetenskaper (creator_code:org_t)

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