Multiplex quantitative PCR for detection of lower respiratory tract infection and meningitis caused by Streptococcus pneumoniae, Haemophilus influenzae and Neisseria meningitidis

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Abdeldaim, Guma,1969-Uppsala universitet,Klinisk bakteriologi,Jonas Blomberg (författare)

Multiplex quantitative PCR for detection of lower respiratory tract infection and meningitis caused by Streptococcus pneumoniae, Haemophilus influenzae and Neisseria meningitidis

Artikel/kapitelEngelska2010

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2010-12-03
Springer Science and Business Media LLC,2010
printrdacarrier

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LIBRIS-ID:oai:DiVA.org:uu-107967
https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-107967URI
https://doi.org/10.1186/1471-2180-10-310DOI
https://gup.ub.gu.se/publication/142313URI

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Språk:engelska
Sammanfattning på:engelska

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Ämneskategori:art swepub-publicationtype

Anmärkningar

Background. Streptococcus pneumoniae and Haemophilus influenzae cause pneumonia and as Neisseria meningitidis they are important agents of meningitis. Although several PCR methods have been described for these bacteria the specificity is an underestimated problem. Here we present a quantitative multiplex real-time PCR (qmPCR) for detection of S. pneumoniae (9802 gene fragment), H. influenzae (omp P6 gene) and N. meningitidis (ctrA gene). The method was evaluated on bronchoalveolar lavage (BAL) samples from 156 adults with lower respiratory tract infection (LRTI) and 31 controls, and on 87 cerebrospinal fluid (CSF) samples from meningitis patients. Results. The analytical sensitivity was not affected by using a combined mixture of reagents and a combined DNA standard (S. pneumoniae/H. influenzae/N. meningitidis) in single tubes. By blood- and BAL-culture and S. pneumoniae urinary antigen test, S. pneumoniae and H. influenzae were aetiological agents in 21 and 31 of the LTRI patients, respectively. These pathogens were identified by qmPCR in 52 and 72 of the cases, respectively, yielding sensitivities and specificities of 95% and 75% for S. pneumoniae, and 90% and 65% for H. influenzae, respectively. When using a cut-off of 105 genome copies/mL for clinical positivity the sensitivities and specificities were 90% and 80% for S. pneumoniae, and 81% and 85% for H. influenzae, respectively. Of 44 culture negative but qmPCR positive for H. influenzae, 41 were confirmed by fucK PCR as H. influenzae. Of the 103 patients who had taken antibiotics prior to sampling, S. pneumoniae and H. influenzae were identified by culture in 6% and 20% of the cases, respectively, and by the qmPCR in 36% and 53% of the cases, respectively. In 87 CSF samples S. pneumoniae and N. meningitidis were identified by culture and/or 16 S rRNA in 14 and 10 samples and by qmPCR in 14 and 10 samples, respectively, giving a sensitivity of 100% and a specificity of 100% for both bacteria. Conclusions. The PCR provides increased sensitivity and the multiplex format facilitates diagnosis of S. pneumoniae, H. influenzae and N. meningitidis and the assay enable detection after antibiotic treatment has been installed. Quantification increases the specificity of the etiology for pneumonia.

Ämnesord och genrebeteckningar

MEDICIN OCH HÄLSOVETENSKAP Medicinska och farmaceutiska grundvetenskaper Mikrobiologi inom det medicinska området hsv//swe
MEDICAL AND HEALTH SCIENCES Basic Medicine Microbiology in the medical area hsv//eng
S. pneumoniae
H. influenzae
N. meningitidis
quantitative multiplex PCR
lower respiratory tract infection
MEDICINE
MEDICIN
Clinical Bacteriology
Klinisk bakteriologi

Biuppslag (personer, institutioner, konferenser, titlar ...)

Strålin, KristofferDepartment of Infectious Diseases, Örebro University Hospital, Örebro (författare)
Korsgaard, JensDepartment of Chest Diseases, Aarhus University Hospital, Aalborg, Denmark (författare)
Blomberg, JonasUppsala universitet,Klinisk virologi(Swepub:uu)jonablom (författare)
Herrmann, BjörnUppsala universitet,Klinisk bakteriologi(Swepub:uu)bjorherr (författare)
Welinder-Olsson, Christina,1959Gothenburg University,Göteborgs universitet,Institutionen för biomedicin, avdelningen för infektionssjukdomar,Institute of Biomedicine, Department of Infectious Medicine(Swepub:gu)xwelch (författare)
Uppsala universitetKlinisk bakteriologi (creator_code:org_t)

Sammanhörande titlar

Ingår i:BMC Microbiology: Springer Science and Business Media LLC10, s. 310-1471-2180

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Av författaren/redakt...: Abdeldaim, Guma, ...; Strålin, Kristof ...; Korsgaard, Jens; Blomberg, Jonas; Herrmann, Björn; Welinder-Olsson, ...

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MEDICIN OCH HÄLSOVETENSKAP: MEDICIN OCH HÄLS ...; och Medicinska och f ...; och Mikrobiologi ino ...

Artiklar i publikationen: BMC Microbiology

Av lärosätet: Uppsala universitet; Göteborgs universitet

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