The invention discloses a novel method for detecting pathogenic microorganisms. The method comprises: a collection step of samples such as
cerebrospinal fluid and
alveolar lavage fluid; a collection steps of
peripheral blood samples;
a DNA extraction step; a
library construction step; a sequencing step; and an analysis step. The
DNA extraction step comprises: taking 400 [mu]l sample for detection,adding reaction reagents of buffer GB,
Lysis Enzyme K,
RNA, Carrier to the sample, conducting reaction at 56 DEG C for 10 minutes to obtain a reaction solution, purifying the reaction solution by
silica gel membrane adsorption column to remove salt ions and organic reagents, and obtaining
DNA with total amount and purity meeting the requirements. The
library construction step comprises: conducting terminal repair,
linker connection,
library amplification and enrichment, purification and
quality control of the
DNA to complete the library construction. The sequencing step comprises: denaturingthe library into a
single strand by NaOH, then binding the library to a sequencing
chip, enriching each
single strand DNA into a cluster by bridge amplification, and obtaining
sequencing data by usingIllumina SBS sequencing method to read the sequence. The analysis step comprises: filtering the
sequencing data, removing the human source and the
linker sequence, and comparing the remaining data with a microbial
reference database.