ABSTRACT
Blood stream infection (BSI),a blood-borne disease caused by microorganisms such as bacteria,fungi,and viruses,can lead to bacteremia,sepsis,and infectious shock,posing a serious threat to human life and health.Identifying the pathogen is central to the precise treatment of BSI.Traditional blood culture is the gold standard for pathogen identification,while it has limitations in clinical practice due to the long time consumption,production of false negative results,etc.Nanopore sequencing,as a new generation of sequencing technology,can rapidly detect pathogens,drug resistance genes,and virulence genes for the optimization of clinical treatment.This paper reviews the current status of nanopore sequencing technology in the diagnosis of BSI.
Subject(s)
Humans , Nanopore Sequencing , Sepsis/diagnosis , Bacteremia/microbiology , Bacteria , Blood Culture/methodsABSTRACT
Objective To investigate the genotype of klebsiella pneumonia strains isolated from eldly inpatients by multiple-locus variable-number tandem-repeat analysis. Methods Totally 184 klebsiella pneumonia strains,isolated from eldly inpatients,were collected,and their genome DNA were extracted. The polymorphism of 7 variable-number tandem-repeat locus in the DNA samples was analyzed by multiple primers polymerase chain reaction and capillary electrophoresis. The clustering analysis of genotyping was carried out with the BioNumerics 5.1 software. Results A total of 139 genotypes were identified in 184 klebsiella pneumonia clinical strains,showing obvious genetic polymorphisms. With clustering analysis of genotypes,all the strains were categorized into three gene clusters (genogroups 1,2,and 3). The genogroup 1 was the biggest cluster,containing 93.06% of the isolated strains. Conclusion There was a predominant cluster in the klebsiella pneumonia strains isolated from eldly inpatients in our center,and the major source of klebsiella pneumonia infection remained the nosocomial infection.
Subject(s)
Aged , Humans , Bacterial Typing Techniques , Cross Infection , Genotype , Genotyping Techniques , Inpatients , Klebsiella pneumoniae , Classification , Minisatellite Repeats , Polymerase Chain Reaction , Polymorphism, GeneticABSTRACT
Bronchiectasis is a chronic lung disorder and a number of bacterial pathogens are involved. However, 30%-40% of sputum and purulent samples in good quality failed to grow any pathogenic bacteria, making it difficult to confirm the pathogen. In this study, we collected bronchoalveolar lavage fluid from a bronchiectasis patient undergoing acute exacerbation, and sent for 16S rDNA pyrosequencing by a 454 GS Junior machine. Metagenomic analysis showed the composition of bacterial community in sample was complex. More than a half of reads (51.3%) were from Pseudomonas aeruginosa. This result was corresponding with the culture result but came out 2 d earlier, which is meaningful for early diagnosis and treatment. The detection with 16S rDNA pyrosequencing technology is more sensitive and rapid than routine culture, and can detect the co-infection or symbiosis in airway, giving us a novel and convenient approach to perform rapid diagnosis.
Subject(s)
Female , Humans , Middle Aged , Bronchiectasis , Microbiology , Bronchoalveolar Lavage Fluid , Chemistry , Microbiology , Early Diagnosis , Metagenome , Genetics , Metagenomics , Methods , Pseudomonas Infections , Microbiology , Pseudomonas aeruginosa , Genetics , RNA, Ribosomal, 16S , Genetics , Time FactorsABSTRACT
<p><b>OBJECTIVE</b>To analyze molecular and evolution characteristics of Salmonella Paratyphi A isolates from 2000 to 2008, China.</p><p><b>METHODS</b>Using pulsed-field gel electrophoresis (PFGE) method with SpeI restriction enzyme, and multilocus sequence typing (MLST) method based on housekeeping genes (aroC, thrA, hisD, purE, sucA, dnaN, hemD, adk, and purA), the genomic variations of 118 Salmonella Paratyphi A isolates from 10 regions during 2000 to 2008 were analyzed.</p><p><b>RESULTS</b>Using PFGE method, 118 Salmonella Paratyphi A isolates were clustered into 32 PFGE patterns, and 5 patterns were predominant (5 isolates or above). However, only 2 MLST types were identified for all isolates with MLST method. Among all Salmonella Paratyphi A isolates, the sequences of housekeeping genes were highly conservative and showed a high degree of cloning.</p><p><b>CONCLUSION</b>For Chinese epidemic Salmonella Paratyphi A isolates during 2000 - 2008, MLST method showed low discrimination power and the MLST method should not be applied to outbreak and epidemiological surveillance of Salmonella Paratyphi A. Currently, nationwide paratyphoid fever epidemics is caused by highly clonal isolates in China. As the time changes, these isolates also accumulate sporadic mutations.</p>