Molecular detection of Streptococcus pneumoniae on dried blood spots from febrile Nigerian children compared to culture

Pui Ying Iroh Tam, Nelmary Hernandez-Alvarado, Mark R. Schleiss, Fatimah Hassan-Hanga, Chuma Onuchukwu, Dominic Umoru, Stephen K Obaro

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Background: Nigeria has one of the highest burdens of pneumococcal disease in the world, but accurate surveillance is lacking. Molecular detection of infectious pathogens in dried blood spots (DBS) is an ideal method for surveillance of infections in resource-limited settings because of its low cost, minimal blood volumes involved, and ease of storage at ambient temperature. Our study aim was to evaluate a Streptococcus pneumoniae real-time polymerase chain reaction (rt-PCR) assay on DBS from febrile Nigerian children on Whatman 903 and FTA filter papers, compared to the gold standard of culture. Methods: Between September 2011 to May 2015, blood was collected from children 5 years of age or under who presented to six hospital study sites throughout northern and central Nigeria with febrile illness, and inoculated into blood culture bottles or spotted onto Whatman 903 or FTA filter paper. Culture and rt-PCR were performed on all samples. Results: A total of 537 DBS specimens from 535 children were included in the study, of which 15 were culture-positive for S. pneumoniae. The rt-PCR assay detected S. pneumoniae in 12 DBS specimens (2.2%). One positive rt-PCR result was identified in a culture-negative specimen from a high-risk subject, and two positive rt-PCR results were negative on repeat testing. Six culture-confirmed cases of S. pneumoniae bacteremia were missed. Compared to culture, the overall sensitivities of Whatman 903 and FTA DBS for detection of S. pneumoniae were 57.1% (95% CI 18.4-90.1%) and 62.5%(95% CI 24.5-91.5%), respectively. Nonspecific amplification was noted in an additional 22 DBS (4.1%). Among these, six were positive for a non-S. pneumoniae pathogen on culture. Conclusions: Rt-PCR was able to detect S. pneumoniae from clinical DBS specimens, including from a culture-negative specimen. Our findings show promise of this approach as a surveillance diagnostic, but also raise important cautionary questions. Several DBS specimens were detected as S. pneumoniae by rt-PCR despite growth of a non-S. pneumoniae pathogen on culture. A precise definition of what constitutes a positive result is required to avoid falsely over-identifying specimens.

Original languageEnglish (US)
Article numbere0152253
JournalPloS one
Volume11
Issue number3
DOIs
StatePublished - Mar 2016

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Streptococcus pneumoniae
fever
Blood
Fever
Polymerase chain reaction
Real-Time Polymerase Chain Reaction
blood
quantitative polymerase chain reaction
Pathogens
Nigeria
Pneumonia
pneumonia
pathogens
monitoring
Assays
bacteremia
burden of disease
blood volume
Bacteremia
Blood Volume

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Tam, P. Y. I., Hernandez-Alvarado, N., Schleiss, M. R., Hassan-Hanga, F., Onuchukwu, C., Umoru, D., & Obaro, S. K. (2016). Molecular detection of Streptococcus pneumoniae on dried blood spots from febrile Nigerian children compared to culture. PloS one, 11(3), [e0152253]. https://doi.org/10.1371/journal.pone.0152253

Molecular detection of Streptococcus pneumoniae on dried blood spots from febrile Nigerian children compared to culture. / Tam, Pui Ying Iroh; Hernandez-Alvarado, Nelmary; Schleiss, Mark R.; Hassan-Hanga, Fatimah; Onuchukwu, Chuma; Umoru, Dominic; Obaro, Stephen K.

In: PloS one, Vol. 11, No. 3, e0152253, 03.2016.

Research output: Contribution to journalArticle

Tam, PYI, Hernandez-Alvarado, N, Schleiss, MR, Hassan-Hanga, F, Onuchukwu, C, Umoru, D & Obaro, SK 2016, 'Molecular detection of Streptococcus pneumoniae on dried blood spots from febrile Nigerian children compared to culture', PloS one, vol. 11, no. 3, e0152253. https://doi.org/10.1371/journal.pone.0152253
Tam PYI, Hernandez-Alvarado N, Schleiss MR, Hassan-Hanga F, Onuchukwu C, Umoru D et al. Molecular detection of Streptococcus pneumoniae on dried blood spots from febrile Nigerian children compared to culture. PloS one. 2016 Mar;11(3). e0152253. https://doi.org/10.1371/journal.pone.0152253
Tam, Pui Ying Iroh ; Hernandez-Alvarado, Nelmary ; Schleiss, Mark R. ; Hassan-Hanga, Fatimah ; Onuchukwu, Chuma ; Umoru, Dominic ; Obaro, Stephen K. / Molecular detection of Streptococcus pneumoniae on dried blood spots from febrile Nigerian children compared to culture. In: PloS one. 2016 ; Vol. 11, No. 3.
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abstract = "Background: Nigeria has one of the highest burdens of pneumococcal disease in the world, but accurate surveillance is lacking. Molecular detection of infectious pathogens in dried blood spots (DBS) is an ideal method for surveillance of infections in resource-limited settings because of its low cost, minimal blood volumes involved, and ease of storage at ambient temperature. Our study aim was to evaluate a Streptococcus pneumoniae real-time polymerase chain reaction (rt-PCR) assay on DBS from febrile Nigerian children on Whatman 903 and FTA filter papers, compared to the gold standard of culture. Methods: Between September 2011 to May 2015, blood was collected from children 5 years of age or under who presented to six hospital study sites throughout northern and central Nigeria with febrile illness, and inoculated into blood culture bottles or spotted onto Whatman 903 or FTA filter paper. Culture and rt-PCR were performed on all samples. Results: A total of 537 DBS specimens from 535 children were included in the study, of which 15 were culture-positive for S. pneumoniae. The rt-PCR assay detected S. pneumoniae in 12 DBS specimens (2.2{\%}). One positive rt-PCR result was identified in a culture-negative specimen from a high-risk subject, and two positive rt-PCR results were negative on repeat testing. Six culture-confirmed cases of S. pneumoniae bacteremia were missed. Compared to culture, the overall sensitivities of Whatman 903 and FTA DBS for detection of S. pneumoniae were 57.1{\%} (95{\%} CI 18.4-90.1{\%}) and 62.5{\%}(95{\%} CI 24.5-91.5{\%}), respectively. Nonspecific amplification was noted in an additional 22 DBS (4.1{\%}). Among these, six were positive for a non-S. pneumoniae pathogen on culture. Conclusions: Rt-PCR was able to detect S. pneumoniae from clinical DBS specimens, including from a culture-negative specimen. Our findings show promise of this approach as a surveillance diagnostic, but also raise important cautionary questions. Several DBS specimens were detected as S. pneumoniae by rt-PCR despite growth of a non-S. pneumoniae pathogen on culture. A precise definition of what constitutes a positive result is required to avoid falsely over-identifying specimens.",
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N2 - Background: Nigeria has one of the highest burdens of pneumococcal disease in the world, but accurate surveillance is lacking. Molecular detection of infectious pathogens in dried blood spots (DBS) is an ideal method for surveillance of infections in resource-limited settings because of its low cost, minimal blood volumes involved, and ease of storage at ambient temperature. Our study aim was to evaluate a Streptococcus pneumoniae real-time polymerase chain reaction (rt-PCR) assay on DBS from febrile Nigerian children on Whatman 903 and FTA filter papers, compared to the gold standard of culture. Methods: Between September 2011 to May 2015, blood was collected from children 5 years of age or under who presented to six hospital study sites throughout northern and central Nigeria with febrile illness, and inoculated into blood culture bottles or spotted onto Whatman 903 or FTA filter paper. Culture and rt-PCR were performed on all samples. Results: A total of 537 DBS specimens from 535 children were included in the study, of which 15 were culture-positive for S. pneumoniae. The rt-PCR assay detected S. pneumoniae in 12 DBS specimens (2.2%). One positive rt-PCR result was identified in a culture-negative specimen from a high-risk subject, and two positive rt-PCR results were negative on repeat testing. Six culture-confirmed cases of S. pneumoniae bacteremia were missed. Compared to culture, the overall sensitivities of Whatman 903 and FTA DBS for detection of S. pneumoniae were 57.1% (95% CI 18.4-90.1%) and 62.5%(95% CI 24.5-91.5%), respectively. Nonspecific amplification was noted in an additional 22 DBS (4.1%). Among these, six were positive for a non-S. pneumoniae pathogen on culture. Conclusions: Rt-PCR was able to detect S. pneumoniae from clinical DBS specimens, including from a culture-negative specimen. Our findings show promise of this approach as a surveillance diagnostic, but also raise important cautionary questions. Several DBS specimens were detected as S. pneumoniae by rt-PCR despite growth of a non-S. pneumoniae pathogen on culture. A precise definition of what constitutes a positive result is required to avoid falsely over-identifying specimens.

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