中国实用儿科杂志

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16SrRNA荧光定量PCR技术检测轮状病毒肠炎患儿肠道菌群变化的临床意义

杨美芬a,黄永坤a,王玉明b,李海林a,甄双平a,戚勤a,张彩营a   

  1. 昆明医学院第一附属医院,a.儿科,b.基因诊断室;云南昆明650032
  • 收稿日期:2006-02-10 修回日期:2006-06-15 出版日期:2006-09-06 发布日期:2006-09-06

Detecting the variation of intestinal microflora in children with rotavirus enteritis by 16SrRNA fluorescent quantificative PCR.

Yang Meifen,Huang YongkunWang Yuming,et al.   

  1. Department of Pediatrics,the First Affiliated Hospital of Kunming Medical College,Kunming 650032,China
  • Received:2006-02-10 Revised:2006-06-15 Online:2006-09-06 Published:2006-09-06

摘要: 目的以轮状病毒(RV)肠炎患儿和健康婴幼儿为研究对象,探讨荧光定量PCR技术在检测肠道菌群变化的实用性和可行性。 方法应用荧光定量PCR技术测定细菌的16SrRNA,对RV肠炎患儿和健康婴幼儿粪便中双歧杆菌、乳酸杆菌和大肠杆菌进行定量检测和分析,并和其他专家用传统方法所获得的结果进行比较。 结果RV肠炎患儿的肠道菌群与健康儿童比较发现肠道中双歧杆菌和乳酸杆菌的数量变化差异有显著性(P<0.05),而大肠杆菌的数量差异无显著性(P>0.05)。其结果与其他文献报道的用细菌培养的方法所得结果一致。 结论RV肠炎患儿肠道中益生菌的数量较正常对照组明显减少。荧光定量PCR技术比传统方法特异性高、敏感性强,更省时和省力,临床上可用此方法对患儿肠道细菌变化进行定量分析。

关键词: 轮状病毒, 肠炎, 菌群, 荧光定量, PCR

Abstract: AbstractObjectiveTo study the variation of intestinal microflora in children with rotavirus enteritis and to understand practicality and feasibility of fluorescent quantificative PCR in the field. MethodsThe bifidobacteria,lactobacillus and escherichia coli in children with rotavirus enteritis and healthy children were detected by 16SrRNAtargeted PCR,and were compared with the result by the conventional method. ResultsCompared with the healthy children,the number of bifidobacteria and lactobacillus appeared significant variation in patients(P<0.05).However,the number of escherichia coli had no significant variation(P>0.05),and the range of bacteria by fluorescent quantificative PCR was nearly equal to the result by the cultivation techniques from other document. ConclusionThe number of probiotics in patients is less than the control.Fluorescent quantificative PCR is a method with high specificity and sensitivity,and it can be applied to the quantification of bacteria in the intestine and saves more time and labor.

Key words: Fluorescent quantificative, PCR , Enteritis, Microflora