北京市朝阳区百日咳鲍特菌分离株药物敏感性和抗原基因型研究

庄吉; 李臻; 侯悦; 贾哲; 胡莹雪; 李阳; 刘洁

doi:10.16766/j.cnki.issn.1674-4152.004363

北京市朝阳区百日咳鲍特菌分离株药物敏感性和抗原基因型研究

doi: 10.16766/j.cnki.issn.1674-4152.004363

庄吉¹,
李臻²,
侯悦³,
贾哲¹,
胡莹雪²,
李阳²,
刘洁^2, ,

1.
包头医学院公共卫生学院，内蒙古包头 014040
2.
北京市朝阳区疾病预防控制中心，北京 100021
3.
首都医科大学附属北京胸科医院转化医学研究中心，北京 101149

基金项目:

北京市自然科学基金-海淀原始创新联合基金资助项目 L202036

详细信息

通讯作者:
刘洁，E-mail：jieliu82@126.com

中图分类号: R378.42
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- 文章访问数: 4
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- 被引次数: 0
出版历程
- 收稿日期: 2025-02-09
- 网络出版日期: 2026-04-11

Study on drug sensitivity and antigenic genotypes of Bordetella pertussis isolates in Chaoyang District, Beijing

ZHUANG Ji¹,
LI Zhen²,
HOU Yue³,
JIA Zhe¹,
HU Yingxue²,
LI Yang²,
LIU Jie^{2
, ,}

1.
Baotou Medical College, School of Public Health, Baotou, Inner Mongolia 014040, China

摘要

摘要: 目的收集百日咳疑似病例的鼻咽拭子，分离并培养百日咳鲍特菌(Bordetella pertussis，Bp)进行药物敏感性和抗原基因型分析，了解2018—2023年北京市朝阳区百日咳鲍特菌分离株的药物敏感性及抗原基因型变化。方法收集百日咳疑似病例鼻咽拭子样本1 200份，利用实时荧光定量PCR系统对鼻咽拭子进行检测，使用E-test试纸法评估Bp对红霉素、阿奇霉素等11种抗菌药物的最低抑菌浓度(MIC)。提取Bp基因组DNA，使用二代测序平台进行全基因组测序，获得抗原基因型和耐药相关位点序列。结果成功培养60株Bp；流行病学调查显示，年龄 < 1岁的患者有29例(48.3%)。59株对红霉素、克林霉素、阿奇霉素、克拉霉素均表现为耐药(MIC≥256 mg/L)，1株对阿奇霉素敏感。此外，60株Bp的抗原序列型别共有2种，为prn1/ptxP1/ptxA1/fim2-1/fim3-1/tcfA2 (5/60，8.3%)、prn2/ptxP3/ptxA1/fim2-1/fim3-1/tcfA2 (55/60，91.7%)。所有菌株的多位点序列分型(MLST)均为ST-2型。所有Bp均发生了23S rRNA A2047G突变。结论 Bp菌株的药物敏感性和抗原基因型均在发生变化，应增加对Bp菌株变异的研究。
- 百日咳 /
- 百日咳鲍特菌 /
- 抗原基因型 /
- 药物敏感性 /
- 红霉素 /
- 大环内酯类
Abstract: Objective To collect nasopharyngeal swabs from suspected pertussis cases and isolate and culture Bordetella pertussis (Bp) for drug sensitivity and antigen genotype studies, thereby elucidating changes in drug sensitivity and antigen genotypes of Bp isolates in Chaoyang District, Beijing, from 2018 to 2023. Methods A total of 1 200 nasopharyngeal swab samples were collected from patients with suspected pertussis. Real-time fluorescent quantitative PCR was used for pathogen detection. Additionally, the E-test strip method was employed to evaluate the minimum inhibitory concentration (MIC) of Bp against 11 antimicrobial agents, including erythromycin and azithromycin. The genomic DNA of Bp isolates was extracted, and whole-genome sequencing was performed using a next-generation sequencing platform to obtain sequences of antigen genotypes and drug resistance-related loci. Results Sixty strains of Bp were successfully cultured. Epidemiological investigation revealed that 29 patients were under 1 year of age, accounting for 48.3%. Of the isolates, 59 strains exhibited resistance to erythromycin, clindamycin, azithromycin, and clarithromycin (MIC≥256 mg/L), while 1 strain was sensitive to azithromycin. The antigen genotypes of the 60 Bp strains were classified into two types: prn1/ptxP1/ptxA1/fim2-1/fim3-1/tcfA2 (5/60, 8.3%) and prn2/ptxP3/ptxA1/fim2-1/fim3-1/tcfA2 (55/60, 91.7%). All strains belonged to multi-locus sequence typing (MLST) type ST-2. A mutation of A2047G in the 23S rRNA gene was identified in all Bp strains. Conclusion The drug sensitivity and antigen genotype of Bp strains are undergoing notable changes, indicating the need for increased research on Bp strain variations.
- Pertussis /
- Bordetella pertussis /
- Antigenic genotype /
- Drug sensitivity /
- Erythromycin /
- Macrolides

HTML全文

图 1 60株Bp分离株的23S rRNA序列对比

注：↓23S rRNA序列中2 047位碱基位点突变；CS为中国Bp疫苗株，Tohama Ⅰ为国际Bp疫苗株。

Figure 1. Comparison of the 23S rRNA sequences of 60 Bp isolates

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表 1 分离菌株药物敏感性实验结果

Table 1. The results of the drug susceptibility testing for the isolated strains

抗生素	药物敏感性实验结果		MIC(μg/mL)
抗生素	敏感率(%)	耐药率(%)	Min~Max	MIC50	MIC90
红霉素	0	100.0(60/60)	≥256	≥256	≥256
阿奇霉素	1.7(1/60)	98.3(59/60)	3~≥256	≥256	≥256
克拉霉素	0	100.0(60/60)	≥256	≥256	≥256
克林霉素	0	100.0(60/60)	≥256	≥256	≥256
头孢呋辛	100.0(60/60)	0	1~32	12	24
头孢曲松	100.0(60/60)	0	0.064~0.250	0.047	0.094
磺胺甲噁唑	100.0(60/60)	0	0.002~6.000	0.094	0.380
美罗培南	100.0(60/60)	0	0.032~0.380	0.094	0.190
哌拉西林	100.0(60/60)	0	＜0.016	＜0.016	＜0.016
氨苄西林	100.0(60/60)	0	0.094~0.500	0.250	0.380
左氧氟沙星	100.0(60/60)	0	0.094~0.750	0.250	0.500

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参考文献(23)

[1]	HASNAIN S, MUNDODAN J, AL BAYAT S, et al. Bordetella pertussis: an agent not to be forgotten in qatar[J]. Qatar Med, 2021, 2021(1): 10. DOI: 10.5339/qmj.2021.10.
[2]	TESSIER E, CAMPBELL H, RIBEIRO S, et al. Impact of the COVID-19 pandemic on bordetella pertussis infections in England[J]. BMC Public Health, 2022, 22(1): 405. DOI: 10.1186/s12889-022-12830-9.
[3]	SZWEJSER-ZAWISLAK E, WILK M M, PISZCZEK P, et al. Evaluation of whole-cell and acellular pertussis vaccines in the context of long-term herd immunity[J]. Vaccines (Basel), 2022, 11(1): 1. DOI: 10.3390/vaccines11010001.
[4]	WANG H, LIU X, CAO X, et al. Global tendency and frontiers of research on pertussis from 2000 to 2023: a bibliometric and visual analysis[J]. Hum Vaccin Immunother, 2024, 20(1): 2392334. DOI: 10.1080/21645515.2024.2392334.
[5]	EVANS N J, ARAKKAL A T, CAVANAUGH J E, et al. The incidence, duration, risk factors, and age-based variation of missed opportunities to diagnose pertussis: a population-based cohort study[J]. Infect Control Hosp Epidemiol, 2023, 44(10): 1629-1636. doi: 10.1017/ice.2023.31
[6]	ANZIANI P, BECKER J, MIGNON C, et al. Deep longitudinal multi-omics analysis of bordetella pertussis cultivated in bioreactors highlights medium starvations and transitory metabolisms, associated to vaccine antigen biosynthesis variations and global virulence regulation[J]. Front Microbiol, 2023, 14: 1036386. DOI: 10.3389/fmicb.2023.1036386.
[7]	《中国感染控制杂志》编辑部. 百日咳诊疗方案(2023年版)[J]. 中国感染控制杂志, 2024, 23(4): 544-546. Editorial Board of the Journal of Infection Control in China. Treatment guidelines for pertussis (2023 edition)[J]. Journal of Infection Control in China, 2024, 23(4): 544-546.
[8]	LINARDOS G, COLTELLA L, RANNO S, et al. Whooping cough cases increase in central italy after COVID-19 pandemic[J]. Antibiotics(Basel), 2024, 13(5): 464. DOI: 10.3390/antibiotics13050464.
[9]	PAYNE M, XU Z, HU D, et al. Genomic epidemiology and multilevel genome typing of bordetella pertussis[J]. Emerg Microbes Infect. 2023, 12(2): 2239945. DOI: 10.1080/22221751.2023.2239945.
[10]	ZHANG J, ZHANG D, WANG X, et al. Macrolide susceptibility and molecular characteristics of bordetella pertussis[J]. J Int Med Res, 2022, 50(2): 3000605221078782. DOI:10.1177/0300060522 1078782.
[11]	PATEL J C, COLE M, RUBIS A B, et al. Differences in pertussis incidence by race and ethnicity in the United States, 2010-2017[J]. Open Forum Infect Dis, 2024, 11(4): ofae177. DOI: 10.1093/ofid/ofae177.
[12]	IVASKA L, BARKOFF A M, MERTSOLA J, et al. Macrolide resistance in bordetella pertussis: current situation and future challenges[J]. Antibiotics (Basel), 2022, 11(11): 1570. DOI: 10.3390/antibiotics11111570.
[13]	FU P, ZHOU J, YANG C, et al. Molecular evolution and increasing macrolide resistance of bordetella pertussis, Shanghai, China, 2016-2022[J]. Emerg Infect Dis, 2023, 30(1): 29-38.
[14]	LI J, LIU L, ZHANG H, et al. Severe problem of macrolides resistance to common pathogens in China[J]. Front Cell Infect Microbiol, 2023, 13: 1181633. DOI: 10.3389/fcimb.2023.1181633.
[15]	LIN X, ZOU J, YAO K, et al. Analysis of antibiotic sensitivity and resistance genes of bordetella pertussis in Chinese children[J]. Medicine (Baltimore), 2021, 100(2): e24090. DOI: 10.1097/MD.0000000000024090.
[16]	王要, 何丽婷, 陈乐园, 等. 复方磺胺甲恶唑治疗儿童百日咳效果观察[J]. 中华全科医学, 2024, 22(4): 614-617. doi: 10.16766/j.cnki.issn.1674-4152.003463 WANG Y, HE L T, CHEN L Y, et al. Effect observation of compound sulfamethoxazole in the treatment of pertussis in children[J]. Chinese Journal of General Practice, 2024, 22(4): 614-617. doi: 10.16766/j.cnki.issn.1674-4152.003463
[17]	中华医学会感染病学分会儿科感染学组. 中国百日咳诊疗与预防指南(2024版)[J]. 中华医学杂志, 2024, 104(15): 1258-1279. Pediatric Infectious Diseases Group of the Chinese Medical Association. Guidelines for diagnosis, treatment, and prevention of pertussis in China (2024 Edition)[J]. Chinese Medical Journal, 2024, 104(15): 1258-1279.
[18]	张娟胜, 张弟强, 林晨, 等. 西安市与上海市百日咳鲍特菌对大环内酯类抗生素的耐药性、耐药相关分子特征及多位点抗原序列分型的比较研究[J]. 西安交通大学学报(医学版), 2022, 43(5): 691-696. ZHANG J S, ZHANG D Q, LIN C, et al. Comparative study on the macrolide antibiotic resistance, resistance-related molecular characteristics, and multilocus antigen sequence typing of bordetella pertussis in Xi ' an and Shanghai[J]. Journal of Xi ' an Jiaotong University(Medical Edition), 2022, 43(5): 691-696.
[19]	李振翠, 马炜森, 王安娜, 等. 广东省两个地区2021—2022年百日咳鲍特菌分离株的抗菌药物敏感性和抗原基因型[J]. 中国疫苗和免疫, 2023, 29(5): 498-502. LI Z C, MA W S, WANG A N, et al. Antimicrobial susceptibility and antigen genotype of bordetella pertussis isolates in two regions of Guangdong province from 2021 to 2022[J]. Chinese Journal of Vaccines and Immunization, 2023, 29(5): 498-502.
[20]	KAMACHI K, KOIDE K, OTSUKA N, et al. Whole-Genome analysis of bordetella pertussis MT27 isolates from school-associated outbreaks: single-nucleotide polymorphism diversity and threshold of the outbreak strains[J]. Microbiol Spectr, 2023, 11(3): e0406522. DOI: 10.1128/spectrum.04065-22.
[21]	MOTALLEBIRAD T, FAZELI H, AZADI D, et al. Determination of capsular serotypes, antibiotic susceptibility pattern, and molecular mechanism of erythromycin resistance among clinical isolates of group b streptococcus in Isfahan, Iran[J]. Adv Biomed Res, 2021, 10: 27. DOI: 10.4103/abr.abr_269_20.
[22]	KOIDE K, YAO S, CHIANG C S, et al. Genotyping and macrolide-resistant mutation of bordetella pertussis in East and South-East Asia[J]. J Glob Antimicrob Resist, 2022, 31: 263-269. doi: 10.1016/j.jgar.2022.10.007
[23]	王院霞, 姚锂凤, 郑巧平, 等. 9株分离自住院患者的CA-MRSA基因型菌株的分子特征及耐药性研究[J]. 诊断学理论与实践, 2021, 20(1): 66-70. WANG Y X, YAO L F, ZHENG Q P, et al. Molecular characteristics and drug resistance of 9 strains of CA-MRSA genotypes isolated from hospitalized patients[J]. Theories and Practices of Diagnostics, 2021, 20(1): 66-70.