Abstract
BackgroundDried blood spot (DBS) is a reliable blood collection method for storing samples at room temperature and easily transporting them. We have previously validated a Real-Time PCR for detection of Streptococcus pneumoniae in DBS. The objective of this study was to apply this methodology for the diagnosis of S. pneumoniae and Haemophilus influenzae b (Hib) in DBS samples of children with pneumonia admitted to two hospitals in Mozambique and Morocco.Methods Ply and wzg genes of S. pneumoniae and bexA gene of Hib, were used as targets of Real-Time PCR. 329 DBS samples of children hospitalized with clinical diagnosis of pneumonia were tested.ResultsReal-Time PCR in DBS allowed for a significant increase in microbiological diagnosis of S. pneumoniae and Hib. When performing blood bacterial culture, only ten isolates of S. pneumoniae and none of Hib were detected (3·0% positivity rate, IC95% 1·4-5·5%). Real-Time PCR from DBS samples increased the detection yield by 4x fold, as 30 S. pneumoniae and 11 Hib cases were detected (12·4% positivity rate, IC95% 9·0-16·5%; P<0·001).ConclusionReal-Time PCR applied in DBS may be a valuable tool for improving diagnosis and surveillance of pneumonia caused by S. pneumoniae or Hib in developing countries.
Highlights
Pneumonia is the main cause of death in children worldwide
We have previously described that the Dried blood spot (DBS) technique enables reproducible storage of samples for identification and serotyping of S. pneumoniae and that its use is an attractive method for preserving samples at room temperature and transporting them [14]
Dried blood spot (DBS) is a useful and easy-to-use method for preserving and transporting blood samples which may have a particular utility in developing countries, where microbiology laboratories are rare and technical expertise is limited
Summary
Pneumonia is the main cause of death in children worldwide. It is estimated that it kills 1·2 million children under five years every year, accounting for 18% of all deaths in this population group [1]. Streptococcus pneumoniae and Haemophilus influenzae b (Hib) are the two principal causes of bacterial pneumonia [1], and major causes of other invasive bacterial diseases These pathogens can be prevented by immunization or treated with low cost antibiotics but it has been estimated that only 30% of children with bacterial pneumonia receive the antibiotics that they need [2]. The introduction of the Hib conjugate vaccine into national childhood immunization programs in the 1990s has resulted in a marked and sustained reduction in the incidence of invasive Hib disease in many countries [3,4,5]. Conclusion: Real-Time PCR applied in DBS may be a valuable tool for improving diagnosis and surveillance of pneumonia caused by S. pneumoniae or Hib in developing countries
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