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[爆炸所致严重烧伤患者的微生物学特征及宏基因组学下一代测序技术在致病微生物检测中的应用]

[Microbiological characteristics of patients with severe burns caused by blast and application of meta- genomics next-generation sequencing in the detection of pathogenic microorganisms].

作者信息

Luo R B, Huang M, Hu H, Zhang R, Han C M

机构信息

Department of Surgery Intensive Care Unit, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou 310009, China.

Department of General Intensive Care Unit, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou 310009, China.

出版信息

Zhonghua Shao Shang Za Zhi. 2021 Oct 20;37(10):946-952. doi: 10.3760/cma.j.cn501120-20201017-00440.

Abstract

To analyze the microbiological characteristics of patients with severe burns caused by blast in different periods and explore the application value of metagenomics next-generation sequencing (mNGS) in detecting pathogenic microorganisms. The retrospective observational study was applied. From June 13 to September 13, 2020, twenty-three patients (21 males and 2 females) with severe burns caused by blast who met the inclusion criteria were admitted to the Second Affiliated Hospital of Zhejiang University School of Medicine, with age of (64±5) years and total burn area of (86±14) % total body surface area. Abbreviated burn severity index (ABSI) score, revised Baux score, acute physiology and chronic health status evaluation (APACHE) Ⅱscore, and sequential organ failure assessment (SOFA) score were counted on admission. Within 7, 8-20 and 21-30 d after admission, the complications, infection source and distribution of pathogenic microorganisms in patients were recorded. The detection of pathogenic microorganisms was analyzed, and the difference in detection efficiency between microbial culture method and mNGS was compared. After admission, the infection of overall source distribution of pathogenic microorganisms in patients was analyzed, and the difference in detection efficiency between microbial culture method and mNGS was compared. Data were statistically analyzed with McNemar and Fisher exact probability test. On admission, ABSI score, revised Baux score, APACHE Ⅱ score and SOFA score were (12.6±2.4), (91±22), (26±4), and (10.3±2.3) respectively. Within 7 d after admission, the main complications of patients were inhalation injury, septic shock, and hypoproteinemia. Patients were mainly infected with pathogenic microorganism on wound, blood stream, and lung. Within 8-20 d after admission, the incidence of septic shock was the highest. The incidence of inhalation injury was significantly lower than that of ≤7 d after admission (<0.01), the main source of infection were wound, lung, and blood stream, and the incidence of wound and blood stream infection were significantly lower than that of ≤7 d after admission (<0.01). Within 21-30 d after admission, the incidences of multiple organ failure and acute respiratory distress syndrome were low, the incidence of inhalation injury was significantly lower than that of ≤7 d after admission (<0.01), and the incidence of septic shock was significantly lower than that of ≤7 d after admission (<0.01) and 8-20 d after admission (<0.01). There were only low bloodstream infections, and the incidence of wound infection was significantly lower than that of ≤7 d after admission (<0.01) and 8-20 d after admission (<0.05), and the incidences of lung and blood stream infection were significantly lower than those of ≤7 d after admission (<0.01). Within ≤7 d after admission, gram-positive bacteria were mainly . Gram-negative bacteria were mainly and . The fungi contained only . Within 8-20 d after admission, was mainly the gram-positive bacteria, and the detection rate of was significantly lower than that of ≤7 d after admission (<0.01). and were the main gram-negative bacteria, and their detection rates were significantly lower than those of ≤7 d after admission (<0.01).There was a new detection of . Within 21-30 d after admission, was the mainly gram-positive bacteria, and the detection rates of and were significantly lower than those of ≤7 d after admission (<0.01). and were still the main gram-negative bacteria, and increased with the extension of time after admission. The detection rate of was significantly higher than that of ≤7 d after admission (<0.01) and 8-20 d after admission (<0.01), and the detection rate of was significantly higher than that of ≤7 d after admission (<0.01). The detection rate of was significantly lower than those of ≤7 d after admission (<0.01) and 8-20 d after admission (<0.01). All were detected. Within ≤7 d and 8-20 d, the consistency between mNGS and bacterial culture was high (=0.659, 0.596). Within 21-30 d after admission, the consistency between mNGS and bacterial culture was moderate (=0.407). In different time periods, the positive test rate of mNGS was basically constant, while that of microbial culture showed a decline with the extension time after admission. Five hundred and six strains of pathogenic microorganisms were isolated from wound, blood, sputum, and indwelling catheter, and , , , were the main pathogenic microorganisms. and were the most common in the wound samples, was more often seen in blood samples while and in sputum samples, and in indwelling catheter samples were the most common. The detection rates of in wound and sputum were significantly higher than those of blood (<0.05 or <0.01) and indwelling catheter (<0.01), respectively. The consistency between the overall results of mNGS and microbial culture were moderate (=0.556). The consistency between mNGS and microbial culture was high in samples of blood and indwelling catheter (=0.631, 0.619), but those were moderate in sputum and wound (=0.558, 0.528). The most common infections of patients with severe burn caused by blast injury were wound infection and blood stream infection. With the extension of time after admission, the main pathogenic bacterial strains of patients changed from , , and to and . mNGS showed a higher positive rate of detecting pathogenic microorganisms than conventional microbial culture.

摘要

分析不同时期爆炸所致重度烧伤患者的微生物学特征,探讨宏基因组二代测序(mNGS)在检测致病微生物中的应用价值。采用回顾性观察性研究。2020年6月13日至9月13日,浙江大学医学院附属第二医院收治符合纳入标准的23例爆炸所致重度烧伤患者(男21例,女2例),年龄(64±5)岁,烧伤总面积(86±14)%体表面积。入院时计算简化烧伤严重程度指数(ABSI)评分、修订版Baux评分、急性生理与慢性健康状况评估(APACHE)Ⅱ评分及序贯器官衰竭评估(SOFA)评分。记录入院后7、8 - 20及21 - 30 d患者的并发症、感染源及致病微生物分布情况。分析致病微生物检测结果,比较微生物培养法与mNGS检测效率的差异。入院后分析患者致病微生物总体来源分布感染情况,比较微生物培养法与mNGS检测效率的差异。数据采用McNemar检验和Fisher确切概率法进行统计学分析。入院时,ABSI评分、修订版Baux评分、APACHEⅡ评分及SOFA评分分别为(12.6±2.4)、(91±22)、(26±4)和(10.3±2.3)。入院后7 d内,患者主要并发症为吸入性损伤、感染性休克及低蛋白血症。患者主要感染部位为创面、血流及肺部。入院后8 - 20 d,感染性休克发生率最高。吸入性损伤发生率显著低于入院后≤7 d(<0.01),主要感染源为创面、肺部及血流,创面及血流感染发生率显著低于入院后≤7 d(<0.01)。入院后21 - 30 d,多器官功能衰竭及急性呼吸窘迫综合征发生率较低,吸入性损伤发生率显著低于入院后≤7 d(<0.01),感染性休克发生率显著低于入院后≤7 d(<0.01)及8 - 20 d(<0.01)。仅血流感染发生率较低,创面感染发生率显著低于入院后≤7 d(<0.01)及8 - 20 d(<0.05),肺部及血流感染发生率显著低于入院后≤7 d(<0.01)。入院后≤7 d,革兰阳性菌主要为 。革兰阴性菌主要为 及 。真菌仅见 。入院后8 - 20 d,主要为革兰阳性菌, 检出率显著低于入院后≤7 d(<0.01)。 及 为主要革兰阴性菌,其检出率显著低于入院后≤7 d(<0.01)。新检出 。入院后21 - 30 d,主要为革兰阳性菌, 及 检出率显著低于入院后≤7 d(<0.01)。 及 仍为主要革兰阴性菌,且随入院后时间延长而增加。 的检出率显著高于入院后≤7 d(<0.01)及8 - 20 d(<0.01), 的检出率显著高于入院后≤7 d(<0.01)。 的检出率显著低于入院后≤7 d(<0.01)及8 - 20 d(<0.01)。所有 均被检出。入院后≤7 d及8 - 20 d,mNGS与细菌培养结果一致性较高( =0.659,0.596)。入院后21 - 30 d,mNGS与细菌培养结果一致性中等( =0.407)。在不同时间段,mNGS阳性检出率基本恒定,而微生物培养阳性检出率随入院后时间延长呈下降趋势。从创面、血液、痰液及留置导管分离出506株致病微生物, 、 、 、 为主要致病微生物。 及 在创面样本中最常见, 在血液样本中较常见, 及 在痰液样本中较常见, 在留置导管样本中最常见。 在创面及痰液中的检出率分别显著高于血液(<0.05或<0.01)及留置导管(<0.01)。mNGS与微生物培养总体结果一致性中等( =0.556)。mNGS与微生物培养在血液及留置导管样本中的一致性较高( =0.631,0.619),但在痰液及创面样本中为中等( =0.558,0.

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