AIM:To implement high-throughput 16S rDNA sequencing to study microbial diversity in the fecal matter of rats with acute lung injury/acute respiratory distress syndrome(ALI/ARDS).METHODS:Intratracheal instillation of ...AIM:To implement high-throughput 16S rDNA sequencing to study microbial diversity in the fecal matter of rats with acute lung injury/acute respiratory distress syndrome(ALI/ARDS).METHODS:Intratracheal instillation of lipopolysaccharide was used to induce ALI,and the pathological changes in the lungs and intestines were observed.D-lactate levels and diamine oxidase(DAO)activities were determined by enzymatic spectrophotometry.The fragments encompassing V4 16S rDNA hypervariable regions were PCR amplified from fecal samples,and the PCR products of V4 were sequenced by Illumina MiSeq.RESULTS:Increased D-lactate levels and DAO activities were observed in the model group(P<0.01).Sequencing results revealed the presence of 3780 and4142 species in the control and model groups,respectively.The percentage of shared species was 18.8419%.Compared with the control group,the model group had a higher diversity index and a lower number of species of Fusobacteria(at the phylum level),Helicobacter and Roseburia(at the genus level)(P<0.01).Differences in species diversity,structure,distribution and composition were found between the control group and early ARDS group.CONCLUSION:The detection of specific bacteria allows early detection and diagnosis of ALI/ARDS.展开更多
基金Grants from the Science and Technology Development Plan of Shandong Province and Taishan Scholar project of Shandong Province
文摘AIM:To implement high-throughput 16S rDNA sequencing to study microbial diversity in the fecal matter of rats with acute lung injury/acute respiratory distress syndrome(ALI/ARDS).METHODS:Intratracheal instillation of lipopolysaccharide was used to induce ALI,and the pathological changes in the lungs and intestines were observed.D-lactate levels and diamine oxidase(DAO)activities were determined by enzymatic spectrophotometry.The fragments encompassing V4 16S rDNA hypervariable regions were PCR amplified from fecal samples,and the PCR products of V4 were sequenced by Illumina MiSeq.RESULTS:Increased D-lactate levels and DAO activities were observed in the model group(P<0.01).Sequencing results revealed the presence of 3780 and4142 species in the control and model groups,respectively.The percentage of shared species was 18.8419%.Compared with the control group,the model group had a higher diversity index and a lower number of species of Fusobacteria(at the phylum level),Helicobacter and Roseburia(at the genus level)(P<0.01).Differences in species diversity,structure,distribution and composition were found between the control group and early ARDS group.CONCLUSION:The detection of specific bacteria allows early detection and diagnosis of ALI/ARDS.
基金This study was supported by grants from the National Natural Science Foundation of China （No. 81372025） and the 2015 Annual Special Cultivation and Development Project for the Technology Innovation Base of the Beijing Key Laboratory of Cardiopulmonary Cerebral Resuscitation （No. Z151100001615056）.
