ABSTRACT
Objective:The whole-genome sequencing and virulence characteristics analysis of a Klebsiella pneumoniae isolate that caused lower limb gas gangrene were performed to provide a reference for the comprehensive understanding of molecular virulence characteristics of K. pneumoniae causing severe community-acquired infection. Methods:The patient was admitted to the emergency department for treatment on March 13, 2018.The main clinical symptoms of the patients were high fever, gas gangrene of the left lower limb, and diabetic ketoacidosis. The pus specimen was collected for the bacterial culture, isolates identification and antimicrobial susceptibility testing. Hypermucoviscous phenotype was detected by string test. The whole genome of the isolate was sequenced and the multi-site sequence typing, capsular serotyping, plasmid characteristics, virulence and antimicrobial resistance genes of the isolate were analyzed. Plasmid curing and conjugation experiments were used to analyze plasmid characteristics. The virulence of the strain was assessed by serum killing and Galleria mellonella lethality assays. A two-sample t-test was used to compare the differences in the lethal dose of 50% (LD 50) between the tested strains and reference strains against the G. mellonella larvae. Results:K. pneumoniae strain KPN41053 was identified, it was only resistant to ampicillin and was negative for hypermucoviscous phenotype. Whole genome sequencing showed that the length of KPN41053 chromosome was 5 377 071 bp, belonging to ST660 type, and the capsular type was K16. A IncFIB(K)/HI1B virulence plasmid (207 506 bp) with a sequence that was highly similar to pLVPK was harbored by KPN41053. The plasmid carried a variety of virulence genes, among which rmpA and rmpA2 were pseudogenes. The plasmid could not be transferred horizontally by conjugation. The variation strain KPN41053_PC was obtained by plasmid curing. Serum killing analysis showed that KPN41053 was serum resistant (Grade 6), and KPN41053_PC was serum intermediately sensitive (Grade 3). The lgLD 50 of KPN41053 had no difference with that of the hypervirulent control strain (ST23-K1 type) ( t=0.32, P=0.765), and was significantly lower than that of KPN41053_PC ( t=5.97, P=0.004). Conclusions:KPN41053 was an atypical hypervirulent K. pneumoniae that belonged to ST660 but without a hypermucoviscous phenotype. The virulence plasmid harbored by KPN41053 was its key virulence factor. Hypervirulent K. pneumoniae can lead to community-acquired gas gangrene in diabetic patient, which deserves clinical attention.
ABSTRACT
Objective:To investigate the detection rate of intestinal colonization of carbapenem-resistant Enterobacterales (CRE) in inpatients, and to analyze the molecular epidemiological characteristics of CRE strains.Methods:This was a prospective study. Stool, rectal swab or perianal swab specimens of 213 inpatients in the surgical intensive care unit (SICU), medical intensive care unit (MICU) and the department of hematology (transplantation ward) in The Second Hospital of Anhui Medical University were collected from March to December, 2019. MacConkey plate containing carbapenems was used to screen CRE strains, and bacteria identification and drug susceptibility test were conducted. Key strains were selected for whole genome sequencing (WGS). Besides, multilocus sequence typing, capsular serotype, drug resistance gene, virulence gene and plasmid carrying characteristics of these strains were analyzed. Using KPN FJ723042 sequence as a reference, the single-nucleotide polymorphism (SNP) of all strains was analyzed.Results:Twenty-three CRE strains were detected, with a detection rate of 10.8%(23/213), which included 15(65.2%) carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates, three (13.0%) Escherichia coli strains, three (13.0%) Enterobacter cloacae strains and two (8.7%) Citrobacter freundii strains. SNP cluster analysis showed that the 15 CRKP strains had two main clonotypes, which were both predominant in SICU. Fifteen strains of CRKP were type ST11-K64. All these strains carried β-lactamase Klebsiella pneumoniae carbapenemase 2 ( blaKPC-2 ), and 12 strains carried regulator of mucoid phenotype gene A2 ( rmpA2) and iucABCD. Conclusions:The detection rate of intestinal colonization of CRE in inpatients is high, and most strains are CRKP of type ST11-K64. These CRKP strains have both multidrug resistance and virulence characteristics, which are risks for hospital transmission.
ABSTRACT
Objective To evaluate the ability of MALDI-TOF MS in identification of the yeast isolated from clinical specimens.Methods 387 strains of yeasts were tested.All the strains were from clinical specimen and identified by VITEK MS and API ID 32C system at the same time.Sequencing of the internal transcribed spacer (ITS) regions of ribosomal DNA was used as the reference method in the analysis of a total of 387 yeast isolates.The accuracy of detected results was analyzed by the X2 test.Results The identification accuracy rates of VITEK MS, API ID 32C were respectively 97.7% and 93.0%.The difference was statistically significant (X2=9.439, P=0.002).There was no statistical significance between MALDI-TOF and ITS sequencing.Conclusion MALDI-TOF is a rapid simple and accurate method which is identified to species level in identification of yeasts.
ABSTRACT
Objective To investigate the clinical features and prognostic factors of nosocomially acquired candidemia.Methods A retrospective analysis was conducted for hospitalized patients with nosocomial candidemia between January 2012 and December 2014 at the First Affiliated Hospital of Anhui Medical University.The univariate and multivariate Logistic regression analyses were used to determine the prognostic factors of candidemia.Results A total of 92 patients were diagnosed with nosocomially acquired candidemia.The most common pathogen was Candida glabrata (C.glabrata,39/92,42.4％),followed by Candida albicans (C.albicans,30/92,32.6％),then Candida krusei (C.krusei,7/92,7.6％),Candida tropicalis (C.tropicalis,5/92,5.4％),Candida parapsilosis (C.parapsilosis,4/92,4.4％) and other Candida spp.(7/92,7.6％).The sensitivity rates of Candida spp.strains against flucytosine,amphotericin B,voriconazole,fluconazole and itraconazol were 100.0％,98.9％,92.4％,82.6％oo and 77.2％,respectively.The 30-day attributable case fatality rate was 13.0％(12/92).Multivariate Logistic regression analyses indicated that presence of central venous catheter (OR=4.833,95％CI:1.010-23.125,P=0.049),invasive mechanical ventilation (OR=6.075,95％CI:1.144-32.257,P=0.034),and receiving hemodialysis (OR =8.367,95 ％ CI:1.390-50.364,P =0.020)were factors independently correlated with increased mortality.Conclusions The pathogens causing nosocomially acquired candidemia are mainly C.glabrata,C.albicans and C.krusei.The drug susceptibility of Candida spp.varies among fluconazole,itraconazol voriconazole.The resistant rates of Candida spp.against voriconazole,fluconazole and itraconazol are different.The presence of central venous catheter,invasive mechanical ventilation and receiving hemodialysis are factors independently correlated with increased mortality.