[1]贾适瑜,王飞燕,杨琳梅,等.宏基因组测序技术检测血流感染样本去宿主干扰效应四种预处理方法的比较[J].现代检验医学杂志,2023,38(04):154-158.[doi:10.3969/j.issn.1671-7414.2023.04.028]
　JIA Shiyu,WANG Feiyan,YANG Linmei,et al.Comparison of the Efficiency of the Four De-host Methods for the Detection of Bacterial Blood Stream Infection Samples with Metagenomic Next Generation Sequencing[J].Journal of Modern Laboratory Medicine,2023,38(04):154-158.[doi:10.3969/j.issn.1671-7414.2023.04.028]
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宏基因组测序技术检测血流感染样本去宿主干扰效应四种预处理方法的比较()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第38卷
期数:
2023年04期
页码:
154-158
栏目:
研究简报·实验技术
出版日期:
2023-07-15

文章信息/Info

Title:
Comparison of the Efficiency of the Four De-host Methods for the Detection of Bacterial Blood Stream Infection Samples with Metagenomic Next Generation Sequencing
文章编号:
1671-7414（2023）04-154-05
作者:
贾适瑜1; 王飞燕1; 杨琳梅1; 耿 洁1; 李 峰1; 高 简2; 周 洲1; 程 军1
（1. 中国医学科学院阜外医院实验诊断中心，北京 100037；2. 云南省阜外心血管病医院检验科，昆明 650102）
Author(s):
JIA Shiyu1;  WANG Feiyan1;  YANG Linmei1;  GENG Jie1;  LI Feng1;  GAO Jian2;  ZHOU Zhou1;  CHENG Jun1
（1.Center of Laboratory Medicine, Fuwai Hospital, Chinese Academy of Medical Sciences, Beijing 100037, China; 2. Department of Clinical Laboratory, Fuwai Cardiovascular Hospital of Yunnan Province, Kunming 650102, China）
关键词:
血流感染; 去宿主方法; 病原微生物; 宏基因组测序
分类号:
Q781；R446
DOI:
10.3969/j.issn.1671-7414.2023.04.028
文献标志码:
A
摘要:
目的　探讨不同去宿主方法对血流感染样本中病原微生物检测的影响。方法　以浓度分别为105，103，102 和10CFU/ml 的模拟金黄色葡萄球菌血流感染样本为研究对象，将检测分为全血组和血浆组，使用宏基因组测序（metagenomic next generation sequencing，mNGS）比较Saponin 法、Wash 法、CD45 法和MolYsis 法四种预处理方法的病原微生物reads 检出数和占比，以评估不同方法的去宿主效率。结果　血流感染病原微生物初始浓度在103CFU/ml以下的微生物检出占比小于0.15%。四种去宿主方法中，Saponin 法reads 检出数最多，病原微生物检出占比最高，血浆组检测结果占比可提升至18% 以上；Wash 法及CD45 法相比直接提取法病原微生物reads 检出数均未显著增加，MolYsis 法的reads 检出数水平有明显增高。血浆组病原微生物reads 检出数多于全血组，去宿主方法可明显提高病原微生物的丰度。结论　对比四种去宿主方法的mNGS 检测结果，Saponin 法可明显去除宿主核酸，提高病原微生物的reads 检出数，尤其血浆组去宿主效果最好。
Abstract:
Objective　To discuss the effects of the different de-host methods for the detection of blood stream infection (BSI) samples. Methods　Staphylococcus aureus was injected into the peripheral BSI samples. And the simulated BSI samples with concentrations of 105, 103, 102, 10CFU/ml, respectively, were as the objects. The detection was divided into the whole blood and the plasma group. Metagenomic next generation sequencing (mNGS) was used to detect the reads and proportion of the nucletc acid of pathogenic microorganism to evaluate the de-host efficiency of four protreatment methods (Saponin method, Wash method, CD45 method and MolYsis method. Results　The detection of pathogenic microorganisms in BSI with an initial concentration below 103CFU/ml was less than 0.15%. Among the four host removal methods, the Saponin method could significantly increase the proportion of microorganisms, especially in the plasma group, which could increase the proportion of pathogen detection results to more than 18%. Compared with the direct extraction method, the Wash method and the CD45 method were not rich pathogenic nucleic acid significantly. The detection level of the reads by MolYsis method was significantly higher than that by direct extraction method. The number of microorganisms detected in the plasma group was higher than that in the whole blood group, and the de-host methods can significantly increase the abundance of pathogenic microorganisms. Conclusion　Comparing the mNGS detection results of the four de-host methods, the Saponin method could significantly remove human nucleic acids and increase the proportion of pathogenic microorganisms, especially in the plasma group, which had the best host removal effect.

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备注/Memo

备注/Memo:
基金项目：云南省心血管病临床医学中心项目（FZX2019-06-01）：二代测序技术检测感染性心内膜炎患者血液中病原菌的应用价值。
作者简介：贾适瑜（1991-），女，硕士研究生，主管技师，研究方向：病原微生物的分子生物学诊断，E-mail：jiashiyu1991@126.com。
通讯作者：程军，E-mail：jackcheng2001@163.com。
更新日期/Last Update: 2023-07-15