Sepsis caused great clinical burden all over the world. This study clarified the value of plasma metagenomic next-generation sequencing (p-mNGS) and blood cell mNGS (bc-mNGS) in sepsis diagnosis and evaluation.

One hundred and fourty-seven blood samples were collected from sepsis patients who met sepsis 3.0 criteria. Blood culture (BC), qPCR, p-mNGS, bc-mNGS and necessary routine assays were conducted. Taking BC and qPCR as reference, diagnosis performance of p-mNGS and bc-mNGS was analyzed. Blood transcriptome was conducted to evaluate the immunological response of patients in groups with different p/bc-mNGS results. Impact of antibiotic use on different methods was also analyzed.

The p-mNGS demonstrated a sensitivity of 100% for bacteria/fungi and 97% for viruses, which was higher than bc-mNGS (88% for bacteria and fungi, 71% for viruses). However, bc-mNGS showed higher concordance with BC results, which indicated that co-mNGS (p-mNGS plus bc-mNGS) protocol increased sensitivity and was helpful to justify viable blood pathogens in sepsis patients. This study showed that p-mNGS(+) & bc-mNGS(+) samples represented more activated immunity response (low expression of interferon-induced genes and high expression of JAK-STAT pathway genes), poorer clinical laboratory indicators (higher Sequential Organ Failure Assessment, higher procalcitonin and higher C-reactive protein) and lower survival rate. This study also proved that the use of broad-spectrum antibiotics affected much less on p/bc-mNGS diagnostic ability than on BC.

This research highlighted the potential value of plasma and blood-cell co-metagenomic sequencing in precise diagnosis and severity evaluation of sepsis patients, which will benefit the management of sepsis patients.