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. 2021 Nov 8;106(2):454-456.
doi: 10.4269/ajtmh.21-0814.

Detection of Pathogens of Acute Febrile Illness Using Polymerase Chain Reaction from Dried Blood Spots

Brian Grundy  1 , Ursula Panzner  2   3   4 , Jie Liu  1 , Hyon Jin Jeon  2   5 , Justin Im  2 , Vera von Kalckreuth  2 , Frank Konings  2 , Gi Deok Pak  2 , Ligia Maria Cruz Espinoza  2 , Abdramane Soura Bassiahi  6 , Nagla Gasmelseed  7 , Raphaël Rakotozandrindrainy  8 , Suzanne Stroup  1 , Eric R Houpt  1 , Florian Marks  2   5   8
Affiliations

Detection of Pathogens of Acute Febrile Illness Using Polymerase Chain Reaction from Dried Blood Spots

Brian Grundy et al. Am J Trop Med Hyg. .

Abstract

Quantitative polymerase chain reaction (qPCR) of dried blood spots (DBS) for pathogen detection is a potentially convenient method for infectious disease diagnosis. This study tested 115 DBS samples paired with whole blood specimens of children and adolescent from Burkina Faso, Sudan, and Madagascar by qPCR for a wide range of pathogens, including protozoans, helminths, fungi, bacteria, and viruses. Plasmodium spp. was consistently detected from DBS but yielded a mean cycle threshold (Ct) 5.7 ± 1.6 higher than that from whole blood samples. A DBS qPCR Ct cutoff of 27 yielded 94.1% sensitivity and 95.1% specificity against the whole blood qPCR cutoff of 21 that has been previously suggested for malaria diagnosis. For other pathogens investigated, DBS testing yielded a sensitivity of only 8.5% but a specificity of 98.6% compared with whole blood qPCR. In sum, direct PCR of DBS had reasonable performance for Plasmodium but requires further investigation for the other pathogens assessed in this study.

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Figures

Figure 1.
Figure 1.
Correlation of whole blood specimens paired with dried blood spots by qPCR for Plasmodium spp. Line of best fit (y = 1.02x + 5.41, R2 = 0.889) of Ct-values, excluding negative values (Ct = 40). Ct = cycle threshold; DBS = dried blood spot; qPCR = quantitative polymerase chain reaction.
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