刘立雍, 张倚玮, 向才鑫, 何利华, 孟凡亮, 刘洁, 张建中, 赵飞. 大环内酯类抗生素耐药肺炎支原体标准菌株的建立与评价[J]. 凯发娱乐加盟代理, 2023, 38(12): 1466-1472. DOI: 10.3784/jbjc.202308070396
引用本文: 刘立雍, 张倚玮, 向才鑫, 何利华, 孟凡亮, 刘洁, 张建中, 赵飞. 大环内酯类抗生素耐药肺炎支原体标准菌株的建立与评价[J]. 凯发娱乐加盟代理, 2023, 38(12): 1466-1472. DOI: 10.3784/jbjc.202308070396
Liu Liyong, Zhang Yiwei, Xiang Caixin, He Lihua, Meng Fanliang, Liu Jie, Zhang Jianzhong, Zhao Fei. Establishment and evaluation of standard strain of macrolide-resistant Mycoplasma pneumonia[J]. Disease Surveillance, 2023, 38(12): 1466-1472. DOI: 10.3784/jbjc.202308070396
Citation: Liu Liyong, Zhang Yiwei, Xiang Caixin, He Lihua, Meng Fanliang, Liu Jie, Zhang Jianzhong, Zhao Fei. Establishment and evaluation of standard strain of macrolide-resistant Mycoplasma pneumonia[J]. Disease Surveillance, 2023, 38(12): 1466-1472. DOI: 10.3784/jbjc.202308070396

大环内酯类抗生素耐药肺炎支原体标准菌株的建立与评价

Establishment and evaluation of standard strain of macrolide-resistant Mycoplasma pneumonia

  • 摘要:
    目的  依据病原微生物菌(毒)种国家标准菌株评价技术标准(WS/T 812—2022)建立大环内酯类抗生素耐药肺炎支原体(Mp)标准菌株,加速我国Mp相关研究进展,推进国家病原微生物资源保藏体系建设。
    方法 通过对菌株进行16S rDNA测序及特异性基因PCR鉴定,确定菌株为Mp菌株。 通过对标准菌株菌落形态观测、基因分型、活性测定、最低抑菌浓度(MIC)值测定、耐药基因检测等明确其生物学特征,并进行菌株稳定性和活性评价。
    结果 标准菌株经16S rDNA测序及特异性基因PCR鉴定为Mp菌种,其连续传代后菌落形态及各种分子生物学特征稳定。 标准菌株对红霉素、克拉霉素和阿奇霉素体外MIC值范围分别为≥256 μg/mL、128 μg/mL~≥256 μg/mL和8 μg/mL~32 μg/mL,在基因水平上表现为23S rDNA基因A2063G突变。 标准菌株在?80 ℃条件下可稳定保存12个月,解冻后需尽快进行增菌培养,避免4 ℃或者室温保存,标准菌株使用时避免超过两次冻融。
    结论 本研究建立并评价了大环内酯类抗生素耐药Mp标准菌株(CHPC 1.13907),为Mp疾病防控、科研以及药物和疫苗的研发等工作提供标准菌株。

     

    Abstract:
    Objective To establish standard strain of macrolide-resistant Mycoplasma pneumoniae (Mp) according to the Technical Requirements for Evaluation of National Standard Strains of Pathogenic Microorganisms(WS/T 812—2022), accelerate the progress of Mp research, and promote the construction of a national pathogenic microbial resource conservation system.
    Methods The Mp standard strain was identified by 16S rDNA sequencing and specific gene PCR detection. The biological characteristics of standard strains were clarified through observation of colony morphology, genotyping, activity determination, MIC value determination and drug resistance gene detection, as well as strain stability and activity evaluation.
    Results The standard strain was identified by 16S rDNA sequencing and specific gene PCR as Mp strain, and its colony morphology and various molecular biological characteristics were stable after successive generations. Minimum inhibitory concentration values of the standard strain in vitro ranged from ≥256 μg/mL, 128 μg/mL to ≥256 μg/mL and 8 μg/mL to 32 μg/mL for erythromycin, clarithromycin and azithromycin, respectively. The standard strain exhibited the 23S rDNA gene A2063G mutation at the genetic level. The standard strain can be stored stably for 12 months at ?80 ℃. After thawing, the standard strain should be incubated as soon as possible, avoiding storage at 4 ℃ or room temperature and avoiding more than two freeze-thaws during use.
    Conclusion This study established and evaluated standard strain of macrolide-resistant Mp(CHPC 1.13907), providing an important material basis for Mp disease prevention and control, scientific research, drug development and vaccine development.

     

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