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目的:了解沈阳地区的肺炎支原体(Mycoplasma pnemoniae,MP)23SrRNA基因的突变现状,为进一步认识突变MP所致肺炎支原体肺炎(MPP)的临床表现及其治疗打下基础。方法:对180例疑似MPP住院患儿咽拭子标本提取MP-DNA应用荧光探针定量PCR(FQ-PCR)技术进行分子鉴定,并于患儿病程7天后采血,应用颗粒凝胶法(PA)进行肺炎支原体抗体测定;对两者均阳性标本应用巢式PCR扩增23SrRNA基因并进行电泳检测及DNA测序分析,将测得序列与基因库中MP标准株基因序列进行比较。结果:180例临床标本荧光探针定量和肺炎支原体抗体均确定阳性52例,应用巢式PCR技术扩增MP-23SrRNA基因,并对扩增产物进行测序,测出45例,45例23SrRNA中均存在2063位A→G的点突变,其中2例为2063位点A、G碱基共存,考虑为敏感株与耐药株共生,突变率100%。结论:沈阳地区MP23SrRNA基因中普遍存在点突变,突变MP与患儿的临床特点有何联系有待于进一步研究。
Objective: To understand the status of 23SrRNA gene mutation in Mycoplasma pnemoniae (MP) in Shenyang and lay a foundation for further understanding of the clinical manifestations and treatment of Mycoplasma pneumoniae pneumonia (MPP) caused by mutant MP. Methods: 180 cases of suspected MPP inpatients with throat swab specimens extracted MP-DNA using fluorescent probe quantitative PCR (FQ-PCR) technology for molecular identification, and 7 days after the course of disease in children with blood, the application of particle gel method (PA ) Were used to detect Mycoplasma pneumoniae antibody. 23S rRNA gene was amplified by nested PCR and tested by electrophoresis and DNA sequencing. The sequence of the 23S rRNA gene was compared with that of the MP standard gene in the gene bank. Results: Quantitative fluorescent probes and antibodies to Mycoplasma pneumoniae were determined in 180 cases of clinical samples. The MP-23S rRNA gene was amplified by nested PCR and the amplified products were sequenced. Among the 45 cases, 45 cases of 23S rRNA There are 2063 A → G point mutations, including 2 cases of 2063 A, G base coexistence, consider the sensitive strains and drug-resistant strains symbiotic mutation rate of 100%. Conclusion: There is a common point mutation in MP23SrRNA gene in Shenyang area. The relationship between MP and clinical features of children needs to be further studied.