BACKGROUND Infectious diseases are still one of the greatest threats to human health,and the etiology of 20%of cases of clinical fever is unknown;therefore,rapid identification of pathogens is highly important.Traditi...BACKGROUND Infectious diseases are still one of the greatest threats to human health,and the etiology of 20%of cases of clinical fever is unknown;therefore,rapid identification of pathogens is highly important.Traditional culture methods are only able to detect a limited number of pathogens and are time-consuming;serologic detection has window periods,false-positive and false-negative problems;and nucleic acid molecular detection methods can detect several known pathogens only once.Three-generation nanopore sequencing technology provides new options for identifying pathogens.CASE SUMMARY Case 1:The patient was admitted to the hospital with abdominal pain for three days and cessation of defecation for five days,accompanied by cough and sputum.Nanopore sequencing of the drainage fluid revealed the presence of orallike bacteria,leading to a clinical diagnosis of bronchopleural fistula.Cefoperazone sodium sulbactam treatment was effective.Case 2:The patient was admitted to the hospital with fever and headache,and CT revealed lung inflammation.Antibiotic treatment for Streptococcus pneumoniae,identified through nanopore sequencing of cerebrospinal fluid,was effective.Case 3:The patient was admitted to our hospital with intermittent fever and an enlarged neck mass that had persisted for more than six months.Despite antibacterial treatment,her symptoms worsened.The nanopore sequencing results indicate that voriconazole treatment is effective for Aspergillus brookii.The patient was diagnosed with mixed cell type classical Hodgkin's lymphoma with infection.CONCLUSION Three-generation nanopore sequencing technology allows for rapid and accurate detection of pathogens in human infectious diseases.展开更多
Cobetia marina is a model proteobacteria in researches on marine biofouling. Its taxonomic nomenclature has been revised many times over the past few decades. To better understand the role of the surface-associated li...Cobetia marina is a model proteobacteria in researches on marine biofouling. Its taxonomic nomenclature has been revised many times over the past few decades. To better understand the role of the surface-associated lifestyle of C. marina and the phylogeny of the family Halomonadaceae, we sequenced the entire genome of C. marina JCM 21022T using single molecule real-time sequencing technology (SMRT) and performed comparative genomics and phylogenomics analyses. The circular chromosome was 4 176 300 bp with an average GC content of 62.44% and contained 3 611 predicted coding sequences, 72 tRNA genes, and 21 rRNA genes. The C. marina JCM 2102U genome contained a set of crucial genes involved in surface colonization processes. The comparative genome analysis indicated the significant differences between C. marina JCM 21022T and Cobetia amphilecti KMM 296 (formerly named C. marina KMM 296) resulted from sequence insertions or deletions and chromosomal recombination. Despite these differences, pan and core genome analysis showed similar gene functions between the two strains. The phylogenomic study of the family Halomonadaceae is relationships were well resolved among every genera Cobetia, Kushneria, Zymobacter, and Halotalea. reported here for the first time. We found that the tested, including Chromohalobacter, Halomonas,展开更多
The gut microbiota of Mongolian hosts has distinctive characteristics due to their meat- and dairyoriented daily diets and unique genotype.The aim of the present study was to investigate the effect of switching from t...The gut microbiota of Mongolian hosts has distinctive characteristics due to their meat- and dairyoriented daily diets and unique genotype.The aim of the present study was to investigate the effect of switching from the typical high protein and fat Mongolian diets to carbohydrate-rich meals composed principally of wheat,rice and naked oats on the host gut microbiota within 3 weeks.Our study took the advantage of the long sequence reads produced by the Pac Bio single molecule real-time sequencing technology to enable the profiling of subjects' gut microbiota communities along the diet intervention to the species precision.We found that the bacterial richness and diversity decreased apparently along the diet intervention.During the diet intervention,the gut microbiota composition displayed no significant difference at phylum level(with major phyla of Firmicutes,Bacteroidetes,Tenericutes and Proteobacteria).The relative abundances of some genera such as Bacteroidetes,Faecalibacterium,Roseburia,Alistipes,Streptococcus,and Oscillospira were significantly altered after the diet switching started.Notably,significant changes were also observed in the proportions of the species Bacteroides dorei,Bacteroides fragilis,Bacteroides thetaiotaomicron,Ruminococcus albus,Ruminococcus faecis,Roseburia faecis and Eubacterium ventriosum.These results have demonstrated that diet and host gut microbiota is closely linked.展开更多
1975年,第一个cDNA的基因组--噬菌体фX174基因组的测序完成,标志着测序时代的开始。1996年以来人类基因组计划的发起极大地推动了测序技术的应用和发展,第二代测序技术也被称为高通量测序技术。而单分子DNA测序技术是最近10年内发展起...1975年,第一个cDNA的基因组--噬菌体фX174基因组的测序完成,标志着测序时代的开始。1996年以来人类基因组计划的发起极大地推动了测序技术的应用和发展,第二代测序技术也被称为高通量测序技术。而单分子DNA测序技术是最近10年内发展起来的新一代的测序技术,称为第三代测序技术,其中包括单分子实时测序(Single molecule real time sequencing,SMRT)、真正单分子测序和单分子纳米孔测序等技术。SMRT测序技术有超长读长、测序周期短、不需要模板扩增和直接检测表观修饰位点等特点,为研究人员提供了新的选择。本文综述了SMRT测序技术的基本原理、性能以及它在微生物16S rRNA基因、微生物全基因组以及微生物宏基因组测序等方面的应用,并分析了SMRT测序技术在环境微生物应用中的优势以及存在的问题。展开更多
基金Supported by Research and Development Funding for Medical and Health Institutions,No.2021YL007.
文摘BACKGROUND Infectious diseases are still one of the greatest threats to human health,and the etiology of 20%of cases of clinical fever is unknown;therefore,rapid identification of pathogens is highly important.Traditional culture methods are only able to detect a limited number of pathogens and are time-consuming;serologic detection has window periods,false-positive and false-negative problems;and nucleic acid molecular detection methods can detect several known pathogens only once.Three-generation nanopore sequencing technology provides new options for identifying pathogens.CASE SUMMARY Case 1:The patient was admitted to the hospital with abdominal pain for three days and cessation of defecation for five days,accompanied by cough and sputum.Nanopore sequencing of the drainage fluid revealed the presence of orallike bacteria,leading to a clinical diagnosis of bronchopleural fistula.Cefoperazone sodium sulbactam treatment was effective.Case 2:The patient was admitted to the hospital with fever and headache,and CT revealed lung inflammation.Antibiotic treatment for Streptococcus pneumoniae,identified through nanopore sequencing of cerebrospinal fluid,was effective.Case 3:The patient was admitted to our hospital with intermittent fever and an enlarged neck mass that had persisted for more than six months.Despite antibacterial treatment,her symptoms worsened.The nanopore sequencing results indicate that voriconazole treatment is effective for Aspergillus brookii.The patient was diagnosed with mixed cell type classical Hodgkin's lymphoma with infection.CONCLUSION Three-generation nanopore sequencing technology allows for rapid and accurate detection of pathogens in human infectious diseases.
基金Supported by the National Natural Science Foundation of China(Nos.41006082,31372517)
文摘Cobetia marina is a model proteobacteria in researches on marine biofouling. Its taxonomic nomenclature has been revised many times over the past few decades. To better understand the role of the surface-associated lifestyle of C. marina and the phylogeny of the family Halomonadaceae, we sequenced the entire genome of C. marina JCM 21022T using single molecule real-time sequencing technology (SMRT) and performed comparative genomics and phylogenomics analyses. The circular chromosome was 4 176 300 bp with an average GC content of 62.44% and contained 3 611 predicted coding sequences, 72 tRNA genes, and 21 rRNA genes. The C. marina JCM 2102U genome contained a set of crucial genes involved in surface colonization processes. The comparative genome analysis indicated the significant differences between C. marina JCM 21022T and Cobetia amphilecti KMM 296 (formerly named C. marina KMM 296) resulted from sequence insertions or deletions and chromosomal recombination. Despite these differences, pan and core genome analysis showed similar gene functions between the two strains. The phylogenomic study of the family Halomonadaceae is relationships were well resolved among every genera Cobetia, Kushneria, Zymobacter, and Halotalea. reported here for the first time. We found that the tested, including Chromohalobacter, Halomonas,
文摘The gut microbiota of Mongolian hosts has distinctive characteristics due to their meat- and dairyoriented daily diets and unique genotype.The aim of the present study was to investigate the effect of switching from the typical high protein and fat Mongolian diets to carbohydrate-rich meals composed principally of wheat,rice and naked oats on the host gut microbiota within 3 weeks.Our study took the advantage of the long sequence reads produced by the Pac Bio single molecule real-time sequencing technology to enable the profiling of subjects' gut microbiota communities along the diet intervention to the species precision.We found that the bacterial richness and diversity decreased apparently along the diet intervention.During the diet intervention,the gut microbiota composition displayed no significant difference at phylum level(with major phyla of Firmicutes,Bacteroidetes,Tenericutes and Proteobacteria).The relative abundances of some genera such as Bacteroidetes,Faecalibacterium,Roseburia,Alistipes,Streptococcus,and Oscillospira were significantly altered after the diet switching started.Notably,significant changes were also observed in the proportions of the species Bacteroides dorei,Bacteroides fragilis,Bacteroides thetaiotaomicron,Ruminococcus albus,Ruminococcus faecis,Roseburia faecis and Eubacterium ventriosum.These results have demonstrated that diet and host gut microbiota is closely linked.
文摘1975年,第一个cDNA的基因组--噬菌体фX174基因组的测序完成,标志着测序时代的开始。1996年以来人类基因组计划的发起极大地推动了测序技术的应用和发展,第二代测序技术也被称为高通量测序技术。而单分子DNA测序技术是最近10年内发展起来的新一代的测序技术,称为第三代测序技术,其中包括单分子实时测序(Single molecule real time sequencing,SMRT)、真正单分子测序和单分子纳米孔测序等技术。SMRT测序技术有超长读长、测序周期短、不需要模板扩增和直接检测表观修饰位点等特点,为研究人员提供了新的选择。本文综述了SMRT测序技术的基本原理、性能以及它在微生物16S rRNA基因、微生物全基因组以及微生物宏基因组测序等方面的应用,并分析了SMRT测序技术在环境微生物应用中的优势以及存在的问题。