人肺炎支原体的分离培养及鉴定

Isolation,culture and identification of Mycoplasma pneumoniae

目的分离培养人肺炎支原体(Mycoplasma pneumoniae,MP)菌株,并进行鉴定。方法通过配制添加抗生素的培养基或采用滤膜过滤等几种预处理方法去除大部分真菌和细菌,再对富集培养时间进行摸索,从住院儿童咽拭子及痰样本中分离培养人MP菌株,通过PCR、基因分型、培养特性、生化特性及血清学特性几方面对分离菌株进行鉴定。结果通过依次添加抗生素、滤膜过滤等几种预处理方法传代能去除大部分真菌和细菌,对咽拭子接种6~8 h、痰样本接种18~24 h后进行传代,最终从978株儿童咽拭子及痰样本中分离到42株人MP菌株,所分离菌株从PCR特性、培养特性、生化特性及血清学特性几方面均鉴定为MP,其中39株为P1-Ⅰ型,3株为P1-Ⅱ型。结论建立了从临床标本中分离培养MP菌株的方法,并成功分离到42株MP菌株,对MP的研究和诊断及MP疫苗研发具有重要的应用价值。

Objective To isolate,culture and identify human Mycoplasma pneumoniae（MP）strains.Methods Mycoplasma pneumoniae strains were isolated from throat swabs and sputum samples of hospitalized children by using medium supplemented with antibiotics or filtration with filtering membrane to remove most of the fungi and bacteria,and the time for culture was optimized.The isolates were identified by PCR,genotyping as well as tests for culture characteristics,biochemical characteristics and serological characteristics.Results Most of the fungi and bacteria were removed by several pretreatment methods,such as sequential addition of antibiotics and filtration with filtering membrane.A total of 42 strains of Mycoplasma pneumoniae isolates were obtained from 978 throat swabs and sputum samples by subculture the throat swabs for 6-8 h and the sputum samples for 18-24 h after inoculation,and identified as MP from PCR characteristics,culture characteristics,biochemical characteristics and serological characteristics,of which 39 were P1 typeⅠand 3 were P1 typeⅡ.Conclusion A method for isolation and culture of MP strains from clinical specimens was developed,and 42 MP strains were successfully isolated,which were of an important application value in the research and diagnosis of MP infection and development of MP vaccine.