宏基因组二代测序与传统病原检测在中性粒细胞减少伴发热白血病患儿中的对比研究

doi:10.12372/jcp.2022.21e1444

Abstract

Abstract:

Objective To investigate the application value of metagenomic next-generation sequencing (mNGS) in leukemia children with febrile neutropenic (FN). Methods The clinical data of leukemia children combined with FN after chemotherapy admitted to the pediatric intensive care unit from January 1, 2018 to July 1, 2021 was retrospectively analyzed. The children were divided into group A (bloodstream infection group) and group B (pulmonary infection group) according to the infection foci, and the efficacy of mNGS and traditional pathogen detection (TPD) in the two groups was compared. Results A total of 56 children (27 boys and 29 girls) were included, with a median age of 4.0 (2.0-7.8) years. There were 39 cases in group A (28 cases of ALL and 11 cases of AML) and 17 cases in group B (10 cases of ALL and 7 cases of AML). The commonest pathogens in group A and B were Aspergillus, Candida and Acinetobacter baumannii. The consistency of positive rate of mNGS and TPD was poor (Kappa=0.039). The mNGS positive rate (80.4%) was significantly higher than TPD (58.9%), and the difference was statistically significant (P=0.023). In group A, the positive rates of mNGS for Aspergillus (35.9% vs. 12.8%), Candida (28.2% vs. 10.3%) and Acinetobacter baumannii (28.2% vs. 7.7%) were higher than those of TPD; in group B, the positive rate of mNGS for Aspergillus (41.2% vs. 5.9%) was higher than that of TPD, and the differences were statistically significant (P<0.05). The mNGS can detect rare pathogens, and the detection rate of mNGS for mixed infection was significantly higher than that of TPD, and the difference was statistically significant (P<0.05). Conclusions In leukemia children with FN who failed in initial antibiotic therapy, the commonest pathogens were Aspergillus, Candida, and Acinetobacter baumannii. As a complementary tool for TPD, mNGS has a higher positive detection rate for common pathogens and outstanding efficacy for mixed infections and rare pathogens.

Key words: metagenomic next-generation sequencing, febrile neutropenic, leukemia, child

KANG Lei, GUO Fang, BAI Xinfeng, XU Meixian. Comparative study of metagenomic next-generation sequencing and traditional pathogen detection in leukemia children with febrile neutropenia[J].Journal of Clinical Pediatrics, 2022, 40(7): 539-544.

Figures/Tables 5

References 22

[1]	Bispo JAB, Pinheiro PS, Kobetz EK. Epidemiology and etiology of leukemia and lymphoma[J]. Cold Spring Harb Perspect Med, 2020, 10(6): a034819. doi: 10.1101/cshperspect.a034819
[2]	Lin X, Wang J, Huang X, et al. Global, regional, and national burdens of leukemia from 1990 to 2017: a systematic analysis of the global burden of disease 2017 study[J]. Aging (Albany NY), 2021, 13(7): 10468-10489.
[3]	Mishra K, Kumar S, Ninawe S, et al. The clinical profile, management, and outcome of febrile neutropenia in acute myeloid leukemia from resource constraint settings[J]. Ther Adv Infect Dis, 2021, 8: 20499361211036592.
[4]	Nucci M. How I treat febrile neutropenia mediterr[J]. Mediterr J Hematol Infect Dis, 2021, 13(1): e2021025. doi: 10.4084/mjhid.2021.025
[5]	Martinez-Nadal G, Puerta-Alcalde P, Gudiol C, et al. Inappropriate empirical antibiotic treatment in high-risk neutropenic patients with bacteremia in the eraof multidrug resistance[J]. Clin Infect Dis, 2020, 70(6): 1068-1074. doi: 10.1093/cid/ciz319 pmid: 31321410
[6]	Gu W, Deng X, Lee M, et al. Rapid pathogen detection by metagenomic next-generation sequencing of infected body fluids[J]. Nat Med, 2021, 27(1): 115-124. doi: 10.1038/s41591-020-1105-z
[7]	马彩霞, 陈镜龙, 陆泳, 等. 肺泡灌洗液宏基因组测序在儿童重症肺炎支原体肺炎混合感染中的诊断价值[J]. 临床儿科杂志, 2020, 38(12): 891-895.
[8]	中华医学会检验医学分会临床微生物学组, 中华医学会微生物学与免疫学分会临床微生物学组, 中国医疗保健国际交流促进会临床微生物与感染分会. 宏基因组高通量测序技术应用于感染性疾病病原检测中国专家共识[J]. 中华检验医学杂志, 2021, 44(2): 107-120.
[9]	Tang W, Zhang Y, Luo C, et al. Clinical Application of metagenomic next-generation sequencing for suspected infections in patients with primary immunodeficiency disease[J]. Front Immunol, 2021, 12: 696403. doi: 10.3389/fimmu.2021.696403
[10]	Kanda Y, Kimura SI, Iino M, et al. D-Index-Guided early antifungal therapy versus empiric antifungal therapy for persistent febrile neutropenia: a randomized controlled noninferiority trial[J]. J Clin Oncol, 2020, 38(8): 815-822.
[11]	中华医学会血液学分会, 中国医师协会血液科医师分会. 中国中性粒细胞缺乏伴发热患者抗菌药物临床应用指南(2020年版)[J]. 中华血液学杂志, 2020, 41(12): 969-978.
[12]	Gu W, Deng X, Lee M, et al. Rapid pathogen detection by metagenomic next-generation sequencing of infected body fluids[J]. Nat Med, 2021, 27(1): 115-124. doi: 10.1038/s41591-020-1105-z
[13]	Pagano L, Caira M, Candoni A, et al. The epidemiology of fungal infections in patients with hematologic malignancies: the SEIFEM-2004 study[J]. Haematologica, 2006, 91(8): 1068-1075.
[14]	Lionakis MS, Iliev ID, Hohl TM. Tobias M Hohl. Immunity against fungi[J]. JCI Insight, 2017, 2(11): e93156. doi: 10.1172/jci.insight.93156
[15]	Lionakis MS, Netea MG, Holland SM. Mendelian genetics of human susceptibility to fungal infection[J]. Cold Spring Harb Perspect Med, 2014, 4(6): a019638. doi: 10.1101/cshperspect.a019638
[16]	Lass-Florl C. Diagnosing fungal infections in haematology patients-another case of less is more in the clinical setting?[J]. Clin Microbiol Infect, 2017, 23(12): 896-897. doi: 10.1016/j.cmi.2017.05.022
[17]	El-Mahallawy HA, Hassan SS, El-Wakil M, et al. Increasing antimierobial resistance monitored in surveillance analysis of blood stream infections in febrile neutropenic pediatric oneology patients[J]. Asian Pac J Cancer Prev, 2015, 16(14): 5691-5695. doi: 10.7314/APJCP.2015.16.14.5691
[18]	Kochanek M, Schalk E, von Bergwelt-Baildon M, et al. Management of sepsis in eutropenic cancer patients: 2018 guidelines from the Infectious Diseases Working Party (AGIHO) and Intensive Care Working Party (iCHOP) of the German Society of Hematology and Medical Oncology (DGHO)[J]. Ann Hematol, 2019, 98(5): 1051-1069. doi: 10.1007/s00277-019-03622-0
[19]	Li N, Cai Q, Miao Q, et al. High-throughput metagenomics for identification of pathogens in the clinical settings[J]. Small Methods, 2021, 5(1): 2000792. doi: 10.1002/smtd.202000792
[20]	Zheng Y, Qiu X, Wang T, et al. The Diagnostic value of metagenomic next-generation sequencing in lower respiratory tract infection[J]. Front Cell Infect Microbiol, 2021, 11: 694756. doi: 10.3389/fcimb.2021.694756
[21]	Fang X, Mei Q, Fan X,et al. Diagnostic value of metagenomic next-generation sequencing for the detection of pathogens in bronchoalveolar lavage fluid in ventilator-associated pneumonia patientsl[J]. Front Microbiol, 2020, 11: 599756. doi: 10.3389/fmicb.2020.599756
[22]	Fontana L, Strasfeld L. Respiratory virus infections of the stem cell transplant recipient and the hematologic malignancy patient[J]. Infect Dis Clin North Am, 2019, 33(2): 523-544. doi: 10.1016/j.idc.2019.02.004

项目	A组(n=39)		统计量	P	B组(n=17)		统计量	P
项目	ALL(n=28)	AML(n=11)	统计量	P	ALL(n=10)	AML(n=7)	统计量	P
男性[n(%)]	16(57.1)	6(54.5)	χ²=0.00	1.000	3(30.0)	2(28.6)	—	0.686¹⁾
年龄 [M(P₂₅~P₇₅)]/岁	5.0 (3.0~7.0)	10.0 (2.0~12.0)	Z=1.33	0.182	3.5 (2.0~4.0)	0.8 (0.7~3.0)	Z=1.88	0.061
脓毒性休克[n(%)]	8(28.6)	1(9.1)	χ²=0.77	0.380	0(0.0)	0(0.0)	—	—
肺出血[n(%)]	3(10.7)	1(9.1)	—	1.000¹⁾	2(20.0)	0(0.0)	—	0.485¹⁾
ARDS[n(%)]	0(0.0)	0(0.0)	—	—	1(10.0)	2(28.6)	—	0.537¹⁾
存活[n(%)]	20(71.4)	10(90.9)	χ²=0.77	0.380	9(90.0)	6(85.7)	—	1.000¹⁾
SOFA评分 [M(P₂₅~P₇₅)]/分	6.0 (4.0~8.0)	7.0 (5.0~8.0)	Z=0.68	0.498	6(3~8)	5(2~8)	Z=0.45	0.656
ANC [M(P₂₅~P₇₅)]×10⁹/L	0.0 (0.0~0.1)	0.0 (0.0~0.1)	Z=1.21	0.225	0.1 (0.0~0.1)	0.1 (0.0~0.2)	Z=0.30	0.768
CRP [M(P₂₅~P₇₅)]/mg·L^-1	146.5 (57.0~231.0)	111.0 (99.0~195.0)	Z=0.19	0.851	62.5 (40.0~152.0)	102.0 (54.0~161.0)	Z=1.08	0.282
PCT [M(P₂₅~P₇₅)]/μg·L^-1	1.5 (0.2~8.3)	0.3 (0.2~1.6)	Z=1.37	0.169	0.8 (0.4~1.4)	0.2 (0.1~1.2)	Z=1.43	0.143
mNGS延迟天数 [M(P₂₅~P₇₅)]/d	10.0 (5.3~13.8)	9.0 (5.0~12.0)	Z=0.71	0.481	13.5 (4.5~24.3)	12.0 (9.0~13.0)	Z=0.49	0.624

病原	A组(n=39)		合计	B组(n=17)		合计
病原	ALL(n=28)	AML(n=11)	合计	ALL(n=10)	AML(n=7)	合计
革兰阴性菌
鲍曼不动杆菌	9(32.1)	3(27.3)	12	3(30.0)	2(28.6)	5
肺炎克雷伯菌	6(21.4)	1(9.1)	7	0(0.0)	1(14.3)	1
铜绿假单胞菌	4(14.3)	0(0.0)	4	0(0.0)	2(28.6)	2
大肠埃希氏菌	2(7.1)	1(9.1)	3	0(0.0)	0(0.0)	0
嗜麦芽窄食单胞菌	3(10.7)	0(0.0)	3	0(0.0)	0(0.0)	0
其他阴性菌	3(10.7)	1(9.1)	4	1(10.0)	1(14.3)	2
革兰阳性菌
肺炎链球菌	3(10.7)	0(0.0)	3	0(0.0)	3(42.9)	3
屎肠球菌	1(3.6)	0(0.0)	1	0(0.0)	3(42.9)	3
金黄色葡萄球菌	1(3.6)	1(9.1)	2	1(10.0)	0(0.0)	1
其他阳性菌	0(0.0)	0(0.0)	0	1(10.0)	0(0.0)	1
真菌
曲霉菌	11(39.3)	4(36.4)	15	6(60.0)	2(28.6)	8
念珠菌	9(32.1)	3(27.3)	12	2(20.0)	2(28.6)	4
肺孢子菌	0(0.0)	1(9.1)	1	0(0.0)	2(28.6)	2
毛霉菌	2(7.1)	1(9.1)	3	0(0.0)	0(0.0)	0
毛孢子菌	0(0.0)	0(0.0)	0	0(0.0)	1(14.3)	1
病毒
EBV	3(10.7)	0(0.0)	3	0(0.0)	1(14.3)	1
CMV	2(7.1)	0(0.0)	2	0(0.0)	0(0.0)	0
腺病毒	1(3.6)	0(0.0)	1	0(0.0)	0(0.0)	0

常见病原	mNGS	TPD	双阳性	双阴性	Kappa	P
A组
曲霉菌	10	1	4	24	0.286	0.012
念珠菌	8	1	3	27	0.294	0.039
鲍曼不动杆菌	9	1	2	27	0.188	0.021
B组
曲霉菌	7	0	1	9	0.131	0.016
鲍曼不动杆菌	1	3	1	12	0.209	0.625

病原体种类	mNGS	TPD	Kappa	P
0	11（19.6）	23（41.1）	0.039	0.023
1	16（28.6）	21（37.5）	0.000	0.424
≥2	29（51.8）	12（21.4）	0.055	0.002

Comparative study of metagenomic next-generation sequencing and traditional pathogen detection in leukemia children with febrile neutropenia

RichHTML

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 5

References 22

Related Articles 15

Metrics

Comments

Recommended 0

[1]	LIN Jinfeng, CHEN Yi, DENG Yan, FENG Ai, HUANG Juan, CHEN Lizhu, XIE Jingqi, NIE Xiaojing. A comparative study on the detection of anti-Nephrin antibodies in children's nephrotic syndrome by chemiluminescence method and cell-based assay [J]. Journal of Clinical Pediatrics, 2026, 44(8): 682-689.
[2]	ZHENG Fangyuan, WANG Miao, DING Mingming, LU Aidong, JIA Yueping, ZENG Huimin, ZHANG Leping. Next-generation sequencing identified gene variants and their prognostic impact in pediatric Philadelphia chromosome positive acute lymphoblastic leukemia [J]. Journal of Clinical Pediatrics, 2026, 44(8): 690-697.
[3]	ZHANG Xiao, FANG Hongjun, KUANG Xiaojun, WANG Lijuan, WU Zhao, JIANG Sha, LI Pei, WU Liwen. Analysis of the clinical and electrophysiological characteristics of atonic seizures in children [J]. Journal of Clinical Pediatrics, 2026, 44(8): 698-704.
[4]	WANG Mengqian, YANG Hui. Case series analysis of 13 intestinal Behcet’s syndrome in children [J]. Journal of Clinical Pediatrics, 2026, 44(8): 705-711.
[5]	WU Xirong, GAO Qi, GUO Siyuan, YIN Qingqin, QIN Qiang, YIN Ju, JIAO Anxia, CHEN Chenghao, WU Di, WANG Bei, DUAN Xiaomin, PENG Yun, XU Baoping, GAO Liwei. Clinical characteristics and prognosis of pulmonary mucormycosis in children with diabetes mellitus [J]. Journal of Clinical Pediatrics, 2026, 44(8): 712-720.
[6]	HAN Yanan, GE Lanlan, CUI Jieyuan. A case report of branchio-oto-renal syndrome caused by EYA1 gene variant with a 6-year follow-up [J]. Journal of Clinical Pediatrics, 2026, 44(8): 721-726.
[7]	SHI Jiayi, GAO Chunlin, JIA Lili, LU Yunyun, SUN Tao, ZHU Xiaodong, XIA Zhengkun, ZHANG Pei. Phenotypic characteristics and prognostic analysis of a pedigree with childhood isolated nephrotic syndrome caused by compound heterozygous mutations in the COQ2 gene [J]. Journal of Clinical Pediatrics, 2026, 44(8): 727-732.
[8]	Rachel Rosen, Osvaldo Borrelli, Christophe Faure, Katja Karrento, Usha Krishnan, Samuel Nurko, Nathalie Rommel, Alan Silverman, Michiel van Wijk, Marc Benninga (Author), Chinese Medical Association Parenteral and Enteral Nutrition Branch Pediatrics Group, The Subspecialty Group of Gastroenterology, The Society of Pediatric, Chinese Medical Association, Gastroenterology Group, Pediatrician Branch, Chinese Medical Doctor Association (Translator). Core interpretation and explanation of Rome Ⅴ diagnostic criteria: upper gastrointestinal disorders of gut-brain interaction in children and adolescents [J]. Journal of Clinical Pediatrics, 2026, 44(7): 595-608.
[9]	Carlo Di Lorenzo, Miguel Saps, Bruno P. Chumpitazi, Shaman Rajindrajith, Annamaria Staiano, Nikhil Thapar, Miranda van Tilburg, Carlos Velasco-Benítez, Arine Vlieger (Author), Chinese Medical Association Parenteral and Enteral Nutrition Branch Pediatrics Group, The Subspecialty Group of Gastroenterology, The Society of Pediatric, Chinese Medical Association, Gastroenterology Group, Pediatrician Branch, Chinese Medical Doctor Association (Translator). Core interpretation and explanation of Rome Ⅴ diagnostic criteria: lower and biliary disorders of gut-brain interaction in children and adolescents [J]. Journal of Clinical Pediatrics, 2026, 44(7): 609-621.
[10]	LI Xufeng, TANG Haoxun, LI Xingjun, ZHANG Lize, GAO Chenqing, ZHAO Shunying, LIU Jianhua. Clinical characteristics, chest CT imaging phenotypes, and their correlation with prognosis in 76 children with Chlamydia pneumoniae pneumonia [J]. Journal of Clinical Pediatrics, 2026, 44(7): 622-627.
[11]	WANG Zhen, SHAO Jingbo, ZHANG Na, XIA Min, ZHU Jiashi, DU Chengkan, LI Hong. Factors influencing long-term recurrence in pediatric acute lymphoblastic leukemia patients with low-risk disease at initial diagnosis: a five-year follow-up analysis of 254 cases from a single center [J]. Journal of Clinical Pediatrics, 2026, 44(7): 628-635.
[12]	GUO Xuemei, XUE Yao, WANG Yongren, RONG Liucheng, FANG Yongjun, LIN Rufeng. Clinical features and prognosis of childhood acute myeloid leukemia with CBFA2T3::GLIS2 fusion [J]. Journal of Clinical Pediatrics, 2026, 44(7): 636-643.
[13]	YANG Lijun, FU Dongxia, CUI Yan, YANG Junmei, ZHANG Liming. Clinical manifestation and genetic analysis of CHARGE syndrome caused by CHD7 gene variation [J]. Journal of Clinical Pediatrics, 2026, 44(7): 644-650.
[14]	HU Yan, YANG Fan, LIANG Aimin, MAO Meng. Identification and management of faltering growth in children: insights from the 13th International Conference on Nutrition and Growth [J]. Journal of Clinical Pediatrics, 2026, 44(6): 485-488.
[15]	LIU Ziyao, WANG Congying, WANG Hongmao, ZHANG Mingming, XU Yingjie, LAI Jianming, NIU Wenquan, LI Xiaohui. Early differentiation of Kawasaki disease with joint symptoms and systemic juvenile idiopathic arthritis with Kawasaki disease-like symptoms: a single-center retrospective study [J]. Journal of Clinical Pediatrics, 2026, 44(6): 489-498.