Abstract
Bloodstream infection (BSI) is a life-threatening condition characterized by the presence of pathogens in the blood. It is associated with increased morbidity and mortality, and has to be treated promptly as mortality increases with every hour of delayed treatment. Therefore, rapid and sensitive diagnosis of BSI is essential. The routine diagnostic method for BSI is blood culture, which can only detect culturable pathogens and takes several days to obtain results. The 16S rRNA gene is present in all bacteria and is commonly used as a target for universal bacterial detection in rapid molecular assays such as PCR. However, molecular detection of the 16S gene is hampered by the large amount of human DNA found in blood samples, making diagnostic results aspecific and less sensitive. We have optimized the selection of PCR primers targeting the 16S rRNA gene to avoid cross-reaction with human DNA background. The developed method increases specificity and sensitivity for pathogen diagnosis, and provides rapid and accurate pathogen detection for rare bacterial DNA in the presence of abundant host DNA.
Highlights
CAG CMG CCG CGG TAA TWC GRG TTT GAT CCT GGC TCA G GCT AAS SGG ATT AGA TAC CC TGG CTC AGR WYG AAC GCT RG GCT CAG ATT GAA CGC TGG CG ACG GCT ACC TTG TTA CGA CTT CTG CTG CCT YCC GTA GAA TTG ACG GGG RCC C ACC GCG GCC KGC TGG C CGA CAG CCA TGC ANC ACC T ACC GCT TGT GCG GGC CC CAA CGC GCA GAA CCT TAC C CGT CAT CCM CAC CTT CCT C CGA CAR CCA TGC ASC ACC T TAC GGR AGG CAG CAG AGA GTT TGA TYM TGG CTC AG AAC GCG AAG AAC CTT AC CGA CGG CCA TGC ANC ACC T AGA GTT TGA TCM TGG C ACC GCG GCK GCT GGC TGC TGC CTC CCG TAG GAG T GAC GGG CGG TGT GTR CA CGG TGT GTA CAA GAC CC TAC GGY TAC CTT GTT ACG ACT T TAC GGT TAC CTT GTT ACG AC AGR GTT YGA TYM TGG CTC AG GGA GGC AGC AGT RRG GAA T GCC AGC AGC CGC GGT AA CCA GCA GCC GCG GTA AT ATT ACC GCG GCT GCT GG TGC CAG CAG CCG CGG TAA TTA CCG CGG CTG CTG GCA C ATT ACC GCG GCT GCT GGC CAK AAA GGA GGT GAT CC CAG GCC TAA CAC ATG CAA GTC AGG ATT AGA TAC CCT AGG GTA TCT AAT CCT GAC TAC CAG GGT ATC TAA T GGA TCC AGA CTT TGA TYM TGG CTC AG AGA GTT TGA TCC TGG CTC AG CCG TCA ATT CCT TTR AGT TT CAA CGC GAA GAA CCT TAC C.
GGT AAG GTT CTT CGC GTT G GAA ACT TAA AKG AAT TG ATG GCT GTC GTC AGC T CCT ACG GGA GGC AGC AG CTA CCA GGG TAT CTA ATC C CAA CGC GAA AAA CCT TAC C TCC TAC GGG AGG CAG CAG T CCT ACG GGN GGC WGC AG GAA GAG TTT GAT CAT GGC TCA G
Summary
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