Methods: Children 5 years of age or under who presented to six hospital study sites throughout northern and central Nigeria with an acute febrile illness (Temp >38.5 oC) associated with difficulty breathing or altered consciousness, were prospectively enrolled from September 2011 through June 2015. Blood was obtained for culture using the automated Bactec incubator system and also spotted onto filter paper. DBS DNA extractions were used as a template in rt-PCR assays with the lytA gene, and RNase P was used as a control to assess the extraction of human cellular DNA.
Results: A total of 270 DBS were evaluated, of which 8 were culture-positive for S. pneumoniae, and 8 were positive by PCR (3.0% each). One culture-confirmed case was missed, and PCR identified one culture-negative specimen from a high-risk group.
Conclusion: Rt-PCR detection of S. pneumoniae in DBS led to an equivalent detection of the pathogen compared to conventional blood culture, and was able to detect S. pneumoniae in culture-negative specimens. PCR assays from DBS are a valuable diagnostic method suitable for conducting surveillance and can provide precise assessments of the effect of conjugate vaccines in resource-limited settings.
P. Y. Iroh Tam,
M. R. Schleiss, None
S. K. Obaro, GENDRIVAX: Scientific Advisor , Travel support
GSK: Investigator , Grant recipient
PFIZER: Investigator , Grant recipient