1.Homology analysis of clinically isolated and colonized linezolid-resistant Enterococcus faecalis strains from a patient
Zhangya PU ; Zhijian YU ; Zhong CHEN ; Xiangbin DENG ; Bing BAI ; Duoyun LI ; Xiaojun LIU ; Xueying HAN ; Fojun LIN ; Qiwen DENG
Chinese Journal of Infection Control 2017;16(4):343-345,350
Objective To study the homology characteristics of clinicaly isolated and colonized linezolid(LZD)-resistant Enterococcus faecalis (E.faecalis) strains from a patient.Methods Ten E.faecalis strains (2 were isolated from urine specimens and 8 were from stool specimens) isolated from a patient with pulmonary infection were performed antimicrobial susceptibility testing, homology of E.faecalis was determined by pulsed-field gel electrophoresis (PFGE).Results Before and after patients received LZD therapy, 2 E.faecalis strains isolated form urine specimens were both resistant to LZD (MICs: 8 mg/mL, 16 mg/mL, respectively), among 8 strains from stool specimens (6 were isolated before therapy, and 2 were isolated after therapy), LZD susceptible, intermediate, and resistant strains were 4, 2, and 2 respectively(MICs: 0.25-12 mg/mL).10 strains of E.faecalis were homologous by PFGE typing.Conclusion In this case, the detection of E.faecalis from urinary tract and intestinal tract is homologous, which suggested that LZD-resistant Enterococcus may be colonized in vivo for a long time, and may be shift to cause bacterial infection.
2.Bloodstream infections caused by Staphylococcus aureus in a university hospital center in Shenzhen, 2008-2015
Jinxin ZHENG ; Hongyan WANG ; Qinzhen XU ; Zhangya PU ; Duoyun LI ; Zhong CHEN ; Xiangbin DENG ; Qiwen DENG ; Zhijian YU
Chinese Journal of Infection and Chemotherapy 2017;17(3):238-244
Objective This study was designed to examine the clinical characteristics of bloodstream infections (BSI) caused by Staphylococcus aureus in a teaching hospital and the risk factors for 30-day mortality.Methods A single center retrospective cohort study was conducted for all the patients with BSI caused by S.aureus between 2008 and 2015.The data of clinical features,microbiology,and 30-day mortality were collected from the database of electronic medical records.Results A total of 121 patients with S.aureus BSI were identified.The prevalence of methicillin-resistant S.aureus (MRSA) was 17.4% (21/121).MRSA BSIs were significantly associated with old age (≥65 years) (P=0.026),hospital acquired infection (P=0.035),respiratory tract infection (P=0.001),polyinfection (P=0.005) and inappropriate initial antibiotic therapy (P=0.001) than methicillin-sensitive S.aureus (MS SA) BSIs.The 30-day mortality was 18.2% (22/121).Both univariate and multivariate analysis suggested that solid tumor (OR,8.932,P=0.004) and septic shock (OR,56.721,P<0.001) were independently associated with the 30-day mortality.Conclusions The present study confirms that solid tumor and septic shock are more important risk factors than MRSA in mortality of patients with S.aureus BSI.
3.Whole genome sequencing for analyzing mutation sites in linezolid-resistant methicillin-resistant Staphylococcus aureus
Weiming YAO ; Zhong CHEN ; Zhangya PU ; Hongyan WANG ; Hang CHENG ; Duoyun LI ; Jinxin ZHENG ; Xiangbin DENG ; Xiaojun LIU ; Qiwen DENG ; Zhijian YU
Chinese Journal of Infection Control 2017;16(1):1-5
Objective To understand genetic mutation sites in linezolid (LZD)-sensitive and inducible resistant strains of methicillin-resistant Staphylococcus aureus (MRSA) using whole-genome sequencing,and realize mutation sites of LZD-resistant gene.Methods MRSA-MS4 with explicit genotype and whole-genome sequences was induced by LZD of different concentration gradients,LZD-resistant strain MRSA-MS4-LZD100 was obtained,minimum inhibitory concentration(MIC) was detected,domain V of 23S rRNA and ribosomal proteins L3/L4 gene in MRSAMS4-LZD100 were amplified by polymerase chain reaction (PCR),the sequenced products obtained the corresponding mutation site in contrast with the wild-type strain;Illumina PE library was constructed through paired-end sequencing by Illumina HiSeq 2000 technique,and whole genome sequencing was completed based on bioinformatics.Results MRAS-MS4-LZD100 strain was induced after 32 passages,MIC of LZD was 96 μg/mL.Sequencing of PCR products indicated the genetic variations were G2447T mutation in multiple copies of domain V of 23S rRNA gene,and Gly113Val mutation in L3 protein respectively;the whole genome of MRSA-MS4-LZD100 contained 2 744 315 bp,annotation of the whole genome found a total of 2 509 genes,11 tRNA-encoding genes and 2 entire rRNA-encoding operons.The data were submitted to the PubMed,and the GeneBank accession number JXMJ00000000 was assigned;a total of 101 SNPs and 6 Small indels were found,16 of 101SNP mutations occurred in exon,of which the variant proteins with anmino acid sequence alterations included IstB ATP binding domain-containing protein,clumping factor A,IS1272 transposase and so on;3 of 6 Small indel mutations occurred in exon,of which the variant proteins with anmino acid sequence alterations included hypothetical protein,30S ribosomal protein S1,and clumping factor A.Conclusion LZD-resistant strain MRSA-MS4-LZD100 was successfully induced by LZD;beside 23S rRNA V domain and ribosomal L3 protein,the other mutant site exist in this resistant strain,which provide some direction for subsequent study of recessive LZD resistance mechanism.