首页|Comparative quantification of human intestinal bacteria based on cPCR and LDR/LCR

Comparative quantification of human intestinal bacteria based on cPCR and LDR/LCR

扫码查看
原文链接
  • NETL
  • NSTL
  • 万方数据
  • 维普
AIM: To establish a multiple detection method based on comparative polymerase chain reaction (cPCR) and ligase detection reaction (LDR)/ligase chain reaction (LCR) to quantify the intestinal bacterial components.METHODS: Comparative quantification of 16S rDNAs from different intestinal bacterial components was used to quantify multiple intestinal bacteria. The 16S rDNAs of different bacteria were amplified simultaneously by cPCR. The LDR/LCR was examined to actualize the genotyping and quantification. Two beneficial (Bifidobacterium, Lactobacillus) and three conditionally pathogenic bacteria (Enterococcus, Enterobacterium and Eubacterium) were used in this detection. With cloned standard bacterial 16S rDNAs, standard curves were prepared to validate the quantitative relations between the ratio of original concentrations of two templates and the ratio of the fluorescence signals of their final ligation products. The internal controls were added to monitor the whole detection flow. The quantity ratio between two bacteria was tested.RESULTS: cPCR and LDR revealed obvious linear correlations with standard DNAs, but cPCR and LCR did not. In the sample test, the distributions of the quantity ratio between each two bacterial species were obtained. There were significant differences among these distributions in the total samples. But these distributions of quantity ratio of each two bacteria remained stable among groups divided by age or sex.CONCLUSION: The detection method in this study can be used to conduct multiple intestinal bacteria genotyping and quantification, and to monitor the human intestinal health status as well.

16s rDNAComparative quantificationComparative polymerase chain reactionIntestinal bacteriaLigase chain reactionLigase detection reaction

Zhou-Rui Tang、Kai Li、Yu-Xun Zhou、Zhen-Xian Xiao、Jun-Hua Xiao、Rui Huang、Guo-Hao Gu

展开 >

College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620, China

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China

Department of Pre-Clinical Medicine School, Soochow University, Suzhou 215123, Jiangsu Province, China

2012

10.3748/wjg.v18.i3.268

世界胃肠病学杂志(英文版)
太原消化病研治中心

世界胃肠病学杂志(英文版)

SCI
影响因子：1.001
ISSN：1007-9327
年,卷(期)：2012.18(3)
  • 10