昆明市10岁以下儿童大环内酯类耐药肺炎支原体情况
| 作者: |
1王霖,
1宾松涛,
1郝芮,
1吴玉芹,
1谭力,
2袁涛,
1叶冬梅,
1胡晓琴,
1邓东佳,
1张婷,
1殷峥,
3施慧,
1李明
1 昆明市儿童医院呼吸内科，昆明 650000 2 云南省第一人民医院妇产科，昆明 650034 3 中国人民解放军东部战区总医院消化内科，南京 210009 |
| 通讯: |
李明
Email: etyyliming@126.com |
| DOI: | 10.3978/j.issn.2095-6959.2022.05.010 |
| 基金: | 云南省应用基础研究-昆医联合专项[2017FE468(-268)]；云南省高层次人才培养计划-学科带头人(D-2018054)。 |
摘要
目的：探讨昆明市10岁以下儿童大环内酯类耐药肺炎支原体感染情况分析。方法：收集昆明市儿童医院社区获得性呼吸道感染患儿500例，其中男237例，女263例，年龄3~10岁。采用聚合酶链反应(polymerase chain reaction，PCR)扩增并测序肺炎支原体耐药株的23S rRNA基因靶区V区域。聚类分析用于预测耐药支原体的变异与传播。结果：在500例呼吸道感染患儿的痰标本中，成功培养出213株，其中药敏菌株70株，中敏菌株96株，耐药菌株47株，耐药菌株检出率为22.06%。用23S rRNA V区引物进行PCR扩增出47株耐药菌株，8株耐药菌株未能扩增，39个耐药菌株的目标条带成功扩增。测序结果显示：16株菌株与标准菌株M129高度同源，16株耐药菌株中，有4株与标准菌株M129相比无基因位点突变。4株含有A2063G转化，3株含有A2064G转化，5株含有A2064C转化。2064点突变的频率(大肠杆菌中为2059)比2063点突变的频率高得多。人耐药肺炎支原体对5种抗生素的表型耐药模式因突变位置不同而不同。突变株A2063G表现出对红霉素和罗红霉素的抗性，突变株A2064G表现出对红霉素和交叉霉素的抗性，突变株A2064C表现出对红霉素的抗性。结论：该研究有助于更好地筛选抗支原体药物，并在临床上更好地治疗对支原体有耐药性的感染患儿。
关键词：
大环内酯类耐药肺炎；儿童；突变株；聚合酶链反应
Macrolide-resistant Mycoplasma pneumoniae in children under 10 years old in Kunming
CorrespondingAuthor: LI Ming Email: etyyliming@126.com
DOI: 10.3978/j.issn.2095-6959.2022.05.010
Foundation: This work was supported by the Joint Program of Applied Basic Research of Yunnan Provincial Department of Science and Technology - Kunming Medical University [2017FE468(-268)] and High-Level Talents Training Program of Yunnan Province - Academic Leader (D-2018054), China.
Abstract
Objective: To investigate the infection of macrolide-resistant Mycoplasma pneumoniae in children under 10 years old in Kunming. Methods: A total of 500 children with community acquired respiratory tract infection in Kunming Children’s Hospital were collected, including 237 boys and 263 girls, aged 3–10 years. The region V of 23S rRNA gene target region of Mycoplasma pneumoniae resistant strain was amplified and sequenced by polymerase chain reaction (PCR). Cluster analysis was used to predict the variation and spread of drug-resistant mycoplasma. Results: Among 500 sputum samples from children with respiratory tract infection, 213 strains were successfully cultured, including 70 drug-sensitive strains, 96 medium-sensitive strains and 47 drug-resistant strains, and the detection rate of drug-resistant strains was 22.06%. Forty-seven drug-resistant strains were amplified by polymerase chain reaction with 23S rRNA V region primers, 8 drug-resistant strains failed to be amplified, and the target bands of 39 drug-resistant strains were successfully amplified. Sequencing results showed that 16 strains were highly homologous to standard strain M129, and 4 of 16 drug-resistant strains had no gene mutation compared with standard strain M129. There are 4 strains containing A2063G transformation, 3 strains containing A2064G transformation and 5 strains containing A2064C transformation. The frequency of 2064 point mutation (2059 in E.coli) was much higher than that of 2063 point mutation. Phenotypic resistance patterns of drug-resistant Mycoplasma pneumoniae to five antibiotics were different due to different mutation positions. The mutant A2063G showed resistance to erythromycin and roxithromycin, the mutant A2064G showed resistance to erythromycin and cross-mycin, and the mutant A2064C showed resistance to erythromycin. Conclusion: This study is helpful to better screen the anti-mycoplasma drugs and better treat the infected children with mycoplasma resistance in clinic.
Keywords：
macrolide resistant pneumonia; children; mutant; polymerase chain reaction