Group B Streptococcus Resistance Gene Analysis And Aspergillus Nidulans Genome Annotation

A revolution in sequencing technologies has led to the reduced cost of genome sequencing.Genome analysis for clinical pathogens using high-throughput sequencing technology produced a comprehensive understanding of the microbial origin,virulence evolution,and pathogenic mechanisms.In this research,we performed genome sequencing for clinical bacterial and fungal pathogens.Combined with genome analysis and phenotypic test,revealed the microbial genetic characterize,anti-resistance and virulence trait.Genome analysis of multi-drug resistance group B StreptococcusObjective Group B Streptococcus(GBS)is a predominant pathogen in neonates and infants.GBS has been intensely investigated in Western countries;however,only a few studies have been carried out in China.In this study,we examined the serotype and antibiotic resistome of clinical strains from China through genome analysis,our results basis for clinical neonatal GBS infection prevention and medical treat.Contents We performed WGS for clinical GBS strains and collected GBS sequencing data from the public database.We assembled 102 GBS genomes and identified the capsular type for all strains.We performed anti-resistant gene screening,and phenotype test revealed the anti-resistant trait and geographical distribution of resistant strains.Methods We obtained the GBS whole-genome sequence through WGS.We assembled sequencing data of 102 strains.We identified the capsular type in molecular methods.We extracted the core-genome SNP data through genome alignment,built population structure through phylogenetic analysis.We screened the resistant genes and compared the structure of resistant element through sequence alignment.Antimicrobial susceptibility test was used to assess the resistant trait.Results The population structure shows 102 isolates are structured into five main clusters,same as the structure in other countries.GBS neonatal infection was most frequently caused by ST17 strains,which also carried more anti-resistant genes than other sequence types.Further phylogenetic analysis revealed ST17 strains formed two clusters in the evolutionary process.ST17 strains most come from Guangzhou and Shenzhen.Two genetic element carrying antibiotic resistant genes inserted in most of the ST17 strains,these strains are resistant to tetracycline,macrolide,and aminoglycosides antibiotics.Conclusion This research is the first population genome analysis of GBS clinical strains collected in China.We investigated serotype of neonatal isolates and resistance trait of all strains.The resistant element carried by some strains can spread through LGT,resulted in more bacteria acquired resistance to antibiotics.We should control the antibiotic uses in the clinical treatment,and develop a GBS vaccine,to prevent the AMR pathogens dissemination under the antibiotic selective pressure.Genome annotation of a clinical Aspergillus nidulans strainObjective Aspergillus nidulans(A.nidulans),is the representative species of Aspergillus.It has been a model organism for eukaryotic genetics over 70 years.A.nidulans is an opportunistic pathogen,which mostly infects CGD patients or immune-comprised person.A healthy person infected with A.nidulans is rare.We performed WGS and high-quality genome annotation for an A.nidulans strain using multiple sequencing technologies.The genome data will support further comparative genomic and phylogenomic analysis.Contents We isolated an A.nidulans strain from a rare case of CNS Aspergillosis,named BJCH M5.We performed molecular identification and morphological characterization.We performed WGS and high-quality genome annotation by transcriptome and proteome data analysis.Methods We combined single molecular real-time(SMRT)sequencing,and high-throughput chromosome conformation capture(Hi-C)sequencing technology finished genome assembly correction of BJCH M5.We integrated transcriptome data and proteome data for genome annotation structure.We utilized many software and database for functional annotation,resulting in high-quality annotation data.Results We found a rare case of CNS aspergillosis for the first time,the patient has a healthy immune function and infected by A.nidulans.Compared with reference strain FGSC A4,clinical strain BJCH M5 has different phenotypes.For further genetic characterization,we performed genome sequencing and annotation.The genome of BJCH M5 includes eight nuclear chromosomes and a circular mitochondrial chromosome.The final annotation results in 10949 genes,in 11470 isoforms,in which 17.6% has a clear functional description.The annotation results were verified by proteome data.Conclusion In this study,we utilized the latest sequencing technology and a comprehensive analyzing pipeline to overcome the difficulties in the past sequencing and annotation of fungal genomes.For the first time,we obtained high-quality genome data of a clinical A.nidulans strain.The genome data laid a solid foundation for the further genomic studies and researches of pathogenic mechanisms.