16S rRNA/DNA荧光定量多聚酶链反应检测肺炎婴幼儿肠道菌群变化  

Changes of Intestinal Flora of Children with Pneumonia Detected by Fluorescent Quantitative 16S rRNA/DNA Targeted Polymerase Chain Reaction Technique

作　　者：王艳丽[1] 刘华[2] 姚萍[3] 鲁萍[1] 徐静[1] 周丽芳[1] 黄永坤[1] 

机构地区：[1]昆明医学院第一附属医院儿科,昆明650032 [2]昆明医学院第一附属医院检验科基因研究室,昆明650032 [3]云南省第二人民医院儿科,昆明650000

出　　处：《实用儿科临床杂志》2008年第10期742-745,共4页Journal of Applied Clinical Pediatrics

基　　金：云南省中青年学术技术带头人后备人才培养计划项目资助(2006py01-18)

摘　　要：目的了解肺炎婴幼儿肠道菌群的变化,探讨16S rRNA/DNA荧光定量多聚酶链反应(PCR)技术在临床细菌定量分析中的可行性和实用性。方法提取23例健康儿童(健康对照组)和23例肺炎患儿(观察组)治疗后粪便标本的细菌DNA,测量和比较二组标本细菌的吸光度(A)260值;采用16SrRNA/DNA荧光定量PCR技术对二组粪便标本中的乳酸杆菌和大肠杆菌进行定量分析和比较。采用SPSS13.0软件进行统计学分析。结果健康对照组和观察组治疗第1、3、7天的粪便标本中细菌的A260值分别为(3381.2±817.2)mg/L、(1643.5±498.4)mg/L、(859.6±165.2)mg/L、(1263.8±337.3)mg/L;健康对照组和观察组治疗第1、3、7天比较,均有显著性差异(Pa<0.01);观察组患儿治疗第1、3、7天和健康对照组的肠道乳酸杆菌数量(拷贝数/g湿便)的对数值分别为(5.19±0.48)、(6.94±0.66)、(6.05±0.57)和(9.13±0.53);大肠杆菌数量的对数值分别是(6.99±1.17)、(7.38±1.08)、(6.76±1.17)、(7.52±0.44),健康对照组和观察组比较,均有显著性差异(Pa<0.01)。结论肺炎患儿随治疗时间长短菌群数量有差异;16S rRNA/DNA荧光定量PCR技术在临床细菌定量分析中是切实可行的。Objective To study the intestinal flora changes of children with pneumonia, and explore the feasibility and practicability of fluorescent quantitative 16S rRNA/DNA targeted polymerase chain reaction（PCR） in quantitative study of bacterium. Methods The bacterial DNA was extracted in stool between 23 healthy children （control group）and 23 children （pneumonia group）with pneumonia after therapy, A260 of bacteria was detected and compared between 2 groups. 16S rRNA/DNA PCR were applied to analyze and compare the bacterial content of lactobacilli and Escherichia coli in stool between control group and pneumonia group. Results Bacterial A260 of stool were respectively （3 381.2 ± 817.2） mg/L in control group, （ 1 643.5 ± 498.4 ） mg/L on therapy group post - treatment first day, （859.6 ± 165.2 ） mg/L on the third day of post - treatment, （ 1 263.8 ± 337.3 ） mg/L on the 7th day. There were significant differences of bacterial A260 between control group and pneumonia group in post - therapy （ Pa 〈 0.01 ）. The logarithms of lactobacilli of stool were respectively （ 5.19 ± 0.48 ） post - therapy group on first day, （ 6.94 ± 0.66） on the third day of post - therapy, （ 6.05 ± 0.57 ） on the 7^th day and （ 9.13 ±0.53 ） in control group. Escherichia coli were respectively （6.99 ± 1.17 ） post - therapy group on the first day, （7.38 ± 1.08 ） on the third day of post - therapy, （6.76 ± 1.17 ） on the 7^th day and （7.52 ± 0.44 ） in control group. The number of Bacillus bifidus and Escherichia coli in stool between control group and pneumonia group had significant differences （ Pa 〈 0.01 ）. Conclusions The sum of microbial population change with the time among children with pneumonia after the treatment;16S rRNA - targeted PCR technique is practicable in study of quantitative analysis of intestinal bacterial flora.

关 键 词：荧光定量 多聚酶链反应 肺炎 肠道菌群 

分 类 号：R725.6[医药卫生—儿科]