The application value of digital PCR and next-generation sequencing technology in the etiological diagnosis of sepsis patients
10.3760/cma.j.cn112309-20240529-00179
- VernacularTitle:数字PCR和二代测序技术在脓毒血症患者病原学诊断中的应用价值
- Author:
Jian SHI
1
;
Jing LEI
;
Zeshi LIU
;
Chaoliang XIONG
;
Tian LI
;
Yan GENG
Author Information
1. 西安交通大学第二附属医院检验科，西安 710004
- Publication Type:Journal Article
- Keywords:
Sepsis;
Digital PCR;
Next-generation sequencing;
Pathogens;
Etiological diagnosis
- From:
Chinese Journal of Microbiology and Immunology
2025;45(3):256-263
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the value of droplet digital PCR (ddPCR) and next-generation sequencing (NGS) technology in the pathogenetic diagnosis of sepsis patients, and to provide a reference basis for the early diagnosis of sepsis.Methods:A retrospective analysis was used to collect the clinical data of 53 patients with suspected sepsis admitted to the intensive care unit (ICU) of the Second Affiliated Hospital of Xi'an Jiaotong University from February to August 2023, and the blood was collected for blood culture, ddPCR and NGS detection simultaneously.Results:After excluding viral infections, the blood culture positive rate was 18.87%(10/53), the ddPCR positive rate was 47.17% (25/53), and the NGS positive rate was 41.51%(22/53). When using the ddPCR detection range as a reference, the ddPCR positivity rate was 98.11% (52/53), while the NGS positivity rate was 84.91%(45/53). There was a statistically significant difference in the positivity rate between the two groups ( P<0.05). Using blood culture as a reference, the sensitivity (60.00% vs 70.00%) and specificity (65.11% vs 69.77%) of ddPCR and NGS were in good agreement. In terms of pathogen detection, NGS had a wider detection range than ddPCR (34 species vs 21 species), and the difference was statistically significant (χ 2=55.000, P<0.001). In terms of time-consumption, blood culture took 66.93 h on average, while ddPCR was faster than NGS (about 4 h vs 20 h ). In terms of antimicrobial resistance (AMR) gene detection, five resistant strains were detected by both ddPCR and NGS, ddPCR detected two AMR genes, namely blaKPC and mecA, while NGS detected five AMR genes. Among them, except for blaKPC which was detected outside the target range in ddPCR, the other four AMR genes were also detected by ddPCR. Conclusions:Compared with blood culture, ddPCR and NGS have good application value in the etiological diagnosis of sepsis. Specifically, ddPCR has a higher detection rate of pathogens and takes less time. On the other hand, NGS has a wider detection range, especially for the discovery of some rare bacteria or pathogens that are difficult to be cultured routinely.
