Abstract：
BACKGROUND: Metagenomic next-generation sequencing (mNGS) of circulating cell-free DNA from plasma is a hypothesis-independent broadband diagnostic method for identification of potential pathogens. So far, it has only been investigated in special risk populations (e.g. patients with neutropenic fever).
OBJECTIVE: To investigate the extent to which mNGS (DISQVER® platform) can be used in routine clinical practice.
METHODS: We collected whole blood specimens for mNGS testing, blood cultures (BC), and pathogen-specific PCR diagnostics. Clinical data and pathogen diagnostics were retrospectively reviewed by an infectious disease expert panel regarding the adjustment of anti-infective therapy.
RESULTS: In 55 selected patients (median age 53 years, 67% male) with heterogeneous diagnoses, a total of 66 different microorganisms and viruses were detected using mNGS (51% viruses, 38% bacteria, 8% fungi, 3% parasites). The overall positivity rate of mNGS was 53% (29/55). Fifty-two out of 66 (79%) potential pathogens detected by mNGS were found in patients with primary or secondary immunodeficiency. The concordance rates of BC and pathogen-specific PCR diagnostics with mNGS testing were 14% (4/28) and 36% (10/28), respectively (p < 0.001). An additional bacterial pathogen (Streptococcus agalactiae) could only be detected by BC. Therapeutic consequences regarding anti-infective therapy were drawn from 23 pathogens (35% of detections), with 18 of these detections occurring in patients with immunodeficiency.
CONCLUSIONS: We conclude that mNGS is a useful diagnostic tool, but should only be performed selectively in addition to routine diagnostics of infectious diseases. The limited number of patients and the retrospective study design do not allow any further conclusions.
OBJECTIVE: To investigate the extent to which mNGS (DISQVER® platform) can be used in routine clinical practice.
METHODS: We collected whole blood specimens for mNGS testing, blood cultures (BC), and pathogen-specific PCR diagnostics. Clinical data and pathogen diagnostics were retrospectively reviewed by an infectious disease expert panel regarding the adjustment of anti-infective therapy.
RESULTS: In 55 selected patients (median age 53 years, 67% male) with heterogeneous diagnoses, a total of 66 different microorganisms and viruses were detected using mNGS (51% viruses, 38% bacteria, 8% fungi, 3% parasites). The overall positivity rate of mNGS was 53% (29/55). Fifty-two out of 66 (79%) potential pathogens detected by mNGS were found in patients with primary or secondary immunodeficiency. The concordance rates of BC and pathogen-specific PCR diagnostics with mNGS testing were 14% (4/28) and 36% (10/28), respectively (p < 0.001). An additional bacterial pathogen (Streptococcus agalactiae) could only be detected by BC. Therapeutic consequences regarding anti-infective therapy were drawn from 23 pathogens (35% of detections), with 18 of these detections occurring in patients with immunodeficiency.
CONCLUSIONS: We conclude that mNGS is a useful diagnostic tool, but should only be performed selectively in addition to routine diagnostics of infectious diseases. The limited number of patients and the retrospective study design do not allow any further conclusions.
摘要：
背景：来自血浆的循环无细胞DNA的宏基因组下一代测序（mNGS）是一种与假设无关的宽带诊断方法，用于鉴定潜在病原体。到目前为止,仅在特殊风险人群中进行了调查（例如中性粒细胞减少症患者）。
目的：研究mNGS（DISQVER®平台）可用于常规临床实践的程度。
方法：我们收集全血标本进行mNGS检测，血培养（BC），和病原体特异性PCR诊断。传染病专家小组就抗感染治疗的调整对临床数据和病原体诊断进行了回顾性审查。
结果：在55名选定的患者中（中位年龄53岁，67%男性)诊断不同，使用mNGS共检测到66种不同的微生物和病毒(51%的病毒，38%的细菌，8%的真菌，3%的寄生虫)。mNGS的总阳性率为53%(29/55)。在原发性或继发性免疫缺陷患者中发现了mNGS检测到的66种潜在病原体中的52种（79％）。BC和病原体特异性PCR诊断与mNGS检测的一致率为14％（4/28）和36％（10/28），分别(p<0.001)。另一种细菌病原体（无乳链球菌）只能通过BC检测到。关于抗感染治疗的治疗后果来自23种病原体（35％的检测），这些检测中有18例发生在免疫缺陷患者中。
结论：我们得出结论，mNGS是一种有用的诊断工具，但除了传染病的常规诊断外，还应选择性地进行。有限的患者数量和回顾性研究设计不允许任何进一步的结论。
目的：研究mNGS（DISQVER®平台）可用于常规临床实践的程度。
方法：我们收集全血标本进行mNGS检测，血培养（BC），和病原体特异性PCR诊断。传染病专家小组就抗感染治疗的调整对临床数据和病原体诊断进行了回顾性审查。
结果：在55名选定的患者中（中位年龄53岁，67%男性)诊断不同，使用mNGS共检测到66种不同的微生物和病毒(51%的病毒，38%的细菌，8%的真菌，3%的寄生虫)。mNGS的总阳性率为53%(29/55)。在原发性或继发性免疫缺陷患者中发现了mNGS检测到的66种潜在病原体中的52种（79％）。BC和病原体特异性PCR诊断与mNGS检测的一致率为14％（4/28）和36％（10/28），分别(p<0.001)。另一种细菌病原体（无乳链球菌）只能通过BC检测到。关于抗感染治疗的治疗后果来自23种病原体（35％的检测），这些检测中有18例发生在免疫缺陷患者中。
结论：我们得出结论，mNGS是一种有用的诊断工具，但除了传染病的常规诊断外，还应选择性地进行。有限的患者数量和回顾性研究设计不允许任何进一步的结论。
