Abstract
Bloodstream infections (BSIs) are often life-threatening, and rapid identification is critical. Here, we developed a TaqMan array card (TAC) assay to detect pathogens in BSI specimens. The TAC included 30 primer/probe pairs targeting 27 species and 3 controls. Reverse transcription and 0.1% blue dextran 2000 increased the TAC assay efficiency. The primer/probe pairs had a limit of detection of 100–102 CFU/mL and a specificity of 100%. For whole blood specimens, the TAC assay showed a sensitivity and specificity of 79.4% and 99.69%, respectively. For blood culture samples, the TAC assay showed a sensitivity and specificity of 100% and 99.67%, respectively. The TAC assay could be a promising method for early detection of bloodstream infection.
Highlights
Severe infectious diseases such as sepsis cause serious morbidity and mortality[1]
Fifty percent Tween 20/Triton X-100 and 0.1% saponin resulted in similar yields of pathogen cells (Fig. 1A), and for easy preparation, 0.1% saponin was used for total nucleic acid (TNA) extraction
Kits 1, 2, 3, and 4 and the benzyl alcohol-guanidine hydrochloride TNA extraction method were tested on blood culture samples showing positive results for S. aureus (G+), E. coli (G−), or C. albicans
Summary
Severe infectious diseases such as sepsis cause serious morbidity and mortality[1]. Immunosuppressants, invasive diagnostic and therapeutic procedures, aging, and multidrug resistance development in pathogenic organisms could increase the incidence of bloodstream infections (BSIs). The current gold standard diagnostic method for BSIs, a blood culture test, is time-consuming and has poor sensitivity for slow growing, intracellular and fastidious microorganisms and antimicrobial-treated patients. Nucleic acid-based methods could shorten the turnaround times, and multiplex base PCR could simultaneously detect multiple pathogens. It has its own limitations, such as the presence of PCR inhibitors and background DNA, low bacterial load, insufficient sensitivity and difficulty in establishing an assay capable of detecting a wide range of pathogens, especially when emergence of a new type or subtype is present. Targeted pathogens S. aureus S. epidermidis S. hominis S. haemolyticus K. oxytoca E. faecalis E. faecium S. pneumoniae S. agalactiae S. pyogenes C. perfringens S. marcescens L. monocytogenes E. coli K. pneumoniae Ab S. maltophilia P. aeruginosa A. xylosoxidans B. cepacia N. meningitides H. influenzae C. neoformans C. glabrata C. tropicalis C. albicans C. parapsilosis
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