Blood samples were collected at enrollment and stored at −80 °C. Within 24 h, the blood samples were transferred to BGI Medical Laboratory Institute (Wuhan, China) for mNGS detection. The mNGS detection was conducted according to the manufacturer’s protocol, which is reported in detail in Supplemental File S2 .
With reference to previous literature, we have formulated the following criteria for positive mNGS results [16 (link)].
For bacteria (mycobacteria excluded), fungi, viruses, and parasites: mNGS identified a microbe (genus level) as positive whose reads were 3-fold greater than that of any other microbes. If only one pathogen was detected, it would be regarded as positive.
For mycobacteria: Mycobacterium tuberculosis was considered positive as long as there was more than 1 read mapped to the species or genus level.
Under the condition that the above criteria were met, the patient’s clinical characteristics were taken into consideration. Parvovirus, human herpes virus 5, etc., were appropriately excluded by clinicians and laboratory staff due to the consideration of laboratory contamination. Other results were treated as positive.
With reference to previous literature, we have formulated the following criteria for positive mNGS results [16 (link)].
For bacteria (mycobacteria excluded), fungi, viruses, and parasites: mNGS identified a microbe (genus level) as positive whose reads were 3-fold greater than that of any other microbes. If only one pathogen was detected, it would be regarded as positive.
For mycobacteria: Mycobacterium tuberculosis was considered positive as long as there was more than 1 read mapped to the species or genus level.
Under the condition that the above criteria were met, the patient’s clinical characteristics were taken into consideration. Parvovirus, human herpes virus 5, etc., were appropriately excluded by clinicians and laboratory staff due to the consideration of laboratory contamination. Other results were treated as positive.
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