• Title/Summary/Keyword: Sequence typing

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Species Transferability of Klebsiella pneumoniae Carbapenemase-2 Isolated from a High-Risk Clone of Escherichia coli ST410

  • Lee, Miyoung;Choi, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.974-981
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    • 2020
  • Sequence type 410 (ST410) of Escherichia coli is an extraintestinal pathogen associated with multi drug resistance. In this study, we aimed to investigate the horizontal propagation pathway of a high-risk clone of E. coli ST410 that produces Klebsiella pneumoniae carbapenemase (KPC). blaKPC-encoding E. coli and K. pneumoniae isolates were evaluated, and complete sequencing and comparative analysis of blaKPC-encoding plasmids from E. coli and K. pneumoniae, antimicrobial susceptibility tests, polymerase chain reaction, multilocus sequence typing, and conjugal transfer of plasmids were performed. Whole-genome sequencing was performed for plasmids mediating KPC-2 production in E. coli and K. pneumoniae clinical isolates. Strains E. coli CPEc171209 and K. pneumoniae CPKp171210 were identified as ST410 and ST307, respectively. CPEc171209 harbored five plasmids belonging to serotype O8:H21, which is in the antimicrobial-resistant clade C4/H24. The CPKp171210 isolate harbored three plasmids. Both strains harbored various additional antimicrobial resistance genes. The IncX3 plasmid pECBHS_9_5 harbored blaKPC-2 within a truncated Tn4401a transposon, which also contains blaSHV-182 with duplicated conjugative elements. This plasmid displayed 100% identity with the IncX3 plasmid pKPBHS_10_3 from the K. pneumoniae CPKp171210 ST307 strain. The genes responsible for the conjugal transfer of the IncX3 plasmid included tra/trb clusters and pil genes coding the type IV pilus. ST410 can be transmitted between patients, posing an elevated risk in clinical settings. The emergence of a KPC-producing E. coli strain (ST410) is concerning because the blaKPC-2-bearing plasmids may carry treatment resistance across species barriers. Transgenic translocation occurs among carbapenem-resistant bacteria, which may spread rapidly via horizontal migration.

Identification of Mating Type Loci and Development of SCAR Marker Genetically Linked to the B3 Locus in Pleurotus eryngii

  • Ryu, Jae-San;Kim, Min Keun;Ro, Hyeon-Su;Kang, Young Min;Kwon, Jin-Hyeuk;Kong, Won-Sik;Lee, Hyun-Sook
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1177-1184
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    • 2012
  • In order to estimate how diverse the mating types in Pleurotus eryngii from different regions are, pairings between monokaryons derived from inter- and intra-groups were done. Sixteen and 15 alleles were identified at loci A and B from the 12 strains. In the P. eryngii KNR2312, widely used for commercial production, four mating loci, A3, A4, B3, and B4, were determined. Those loci, except A3, were found in 4 strains out of 12 strains. To improve breeding efficiency, especially in mating type determination, RAPD and BSA were performed to screen for a mating type specific marker. The SCAR marker 13-$2_{2100}$ was developed based on the RAPD-derived sequence typing B3 locus. The sequence analysis of 13-$2_{2100}$ revealed that it contained a conserved domain, the STE3 super-family, and consensus sequences like the TATA box and GC box. It seems likely that the SCAR marker region is a part of the pheromone receptor gene.

Molecular Characterization of Burkholderia cepacia Complex Isolates Causing Bacterial Fruit Rot of Apricot

  • Li, Bin;Fang, Yuan;Zhang, Guoqing;Yu, Rongrong;Lou, Miaomiao;Xie, Guanlin;Wang, Yanli;Sun, Guochang
    • The Plant Pathology Journal
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    • v.26 no.3
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    • pp.223-230
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    • 2010
  • The Burkholderia cepacia complex isolates causing bacterial fruit rot of apricot were characterized by speciesspecific PCR tests, recA-HaeIII restriction fragment length polymorphism (RFLP) assays, rep-PCR genomic fingerprinting, recA gene sequencing, and multilocus sequence typing (MLST) analysis. Results indicated that the isolates Bca 0901 and Bca 0902 gave positive amplifications with primers specific for B. vietnamiensis while the two bacterial isolates showed different recA-RFLP and rep-PCR profiles from those of B. vietnamiensis strains. In addition, the two bacterial isolates had a higher proteolytic activity compared with that of the non-pathogenic B. vietnamiensis strains while no cblA and esmR marker genes were detected for the two bacterial isolates and B. vietnamiensis strains. The two bacterial isolates were identified as Burkholderia seminalis based on recA gene sequence analysis and MLST analysis. Overall, this is the first characterization of B. seminalis that cause bacterial fruit rot of apricot.

Uropathogenic Escherichia coli ST131 in urinary tract infections in children

  • Yun, Ki Wook;Lee, Mi-Kyung;Kim, Wonyong;Lim, In Seok
    • Clinical and Experimental Pediatrics
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    • v.60 no.7
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    • pp.221-226
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    • 2017
  • Purpose: Escherichia coli sequence type (ST) 131, a multidrug-resistant clone causing extraintestinal infections, has rapidly become prevalent worldwide. However, the epidemiological and clinical features of pediatric infections are poorly understood. We aimed to explore the characteristics of ST131 Escherichia coli isolated from Korean children with urinary tract infections. Methods: We examined 114 uropathogenic E. coli (UPEC) isolates from children hospitalized at Chung-Ang University Hospital between 2011 and 2014. Bacterial strains were classified into STs by partial sequencing of seven housekeeping genes (adk, fumC, gyrB, icd, mdh, purA, and recA). Clinical characteristics and antimicrobial susceptibility were compared between ST131 and non-ST131 UPEC isolates. Results: Sixteen UPEC isolates (14.0%) were extended-spectrum ${\beta}-lactamase$ (ESBL)-producers; 50.0% of ESBL-producers were ST131 isolates. Of all the isolates tested, 13.2% (15 of 114) were classified as ST131. There were no statistically significant associations between ST131 and age, sex, or clinical characteristics, including fever, white blood cell counts in urine and serum, C-reactive protein, radiologic abnormalities, and clinical outcome. However, ST131 isolates showed significantly lower rates of susceptibility to cefazolin (26.7%), cefotaxime (40.0%), cefepime (40.0%), and ciprofloxacin (53.3%) than non-ST131 isolates (65.7%, 91.9%, 92.9%, and 87.9%, respectively; P<0.001 for all). ESBL was more frequently produced in ST131 (53.3%) than in non-ST131 (8.1%) isolates (P<0.01). Conclusion: ST131 E. coli isolates were prevalent uropathogens in children at a single medical center in Korea between 2011 and 2014. Although ST131 isolates showed higher rates of antimicrobial resistance, clinical presentation and outcomes of patients were similar to those of patients infected with non-ST131 isolates.

Genetic Organization of ascB-dapE Internalin Cluster Serves as a Potential Marker for Listeria monocytogenes Sublineages IIA, IIB, and IIC

  • Chen, Jianshun;Fang, Chun;Zhu, Ningyu;Lv, Yonghui;Cheng, Changyong;Bei, Yijiang;Zheng, Tianlun;Fang, Weihuan
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.575-584
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    • 2012
  • Listeria monocytogenes is an important foodborne pathogen that comprises four genetic lineages: I, II, III, and IV. Of these, lineage II is frequently recovered from foods and environments and responsible for the increasing incidence of human listeriosis. In this study, the phylogenetic structure of lineage II was determined through sequencing analysis of the ascB-dapE internalin cluster. Fifteen sequence types proposed by multilocus sequence typing based on nine housekeeping genes were grouped into three distinct sublineages, IIA, IIB, and IIC. Organization of the ascB-dapE internalin cluster could serve as a molecular marker for these sublineages, with inlGHE, inlGC2DE, and inlC2DE for IIA, IIB, and IIC, respectively. These sublineages displayed specific genetic and phenotypic characteristics. IIA and IIC showed a higher frequency of recombination (${\rho}/{\theta}$). However, recombination events had greater effect (r/m) on IIB, leading to its high nucleotide diversity. Moreover, IIA and IIB harbored a wider range of internalin and stress-response genes, and possessed higher nisin tolerance, whereas IIC contained the largest portion of low-virulent strains owing to premature stop codons in inlA. The results of this study indicate that IIA, IIB, and IIC might occupy different ecological niches, and IIB might have a better adaptation to a broad range of environmental niches.

A Foodborne Outbreak of Staphylococcus aureus Associated with Fried Chicken in Republic of Korea

  • Hyeon, Ji-Yeon;Chung, Gyung-Tae;Bing, Sun-Hye;Kwon, Kyung-Sook;Lee, Hyeon-Hee;Kim, Soo-Jin;Jeon, Se-Eun;Kang, Yeon-Ho;Kim, Junyoung
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.85-87
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    • 2013
  • An outbreak of Staphylococcus aureus infections occurred in a university with an enrollment of 80 students in the city of Daejon, Republic of Korea. All nine S. aureus isolates from patients (n = 7), staff members (n = 1), and the fried chicken served as the lunch (n = 1) harbored the enterotoxin A gene and showed an identical antibioticresistant profile, PFGE banding pattern (STAS16.001), and sequence type, ST 6. These results suggested that the outbreak was associated with eating the fried chicken that had been handled by an infected staff member. This case report demonstrated a practical approach to identifying the source and transmission of an infection.

Genotyping of the causative Leptospira in symptomatic dogs in Thailand

  • Paungpin, Weena;Chaiwattanarungruengpaisan, Somjit;Mongkolphan, Chalisa;Wiriyarat, Witthawat;Thongdee, Metawee
    • Korean Journal of Veterinary Research
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    • v.60 no.1
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    • pp.1-7
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    • 2020
  • We investigated the genotypes of Leptospira spp. detected in symptomatic dogs in Thailand. During April to December 2012, 6 out of 41 client-owned dogs were diagnosed with leptospirosis based on polymerase chain reaction tests. All of the infected dogs showed clinical symptoms related to leptospirosis. Direct genotyping of the causative agent of the canine leptospirosis was conducted from the archival DNA samples extracted from urine or blood of those 6 infected dogs. Sequencing of the partial 16S rRNA and lipL32 genes from all samples identified Leptospira (L.) interrogans as the infecting species. Multilocus sequence typing tests were successful for 2 out of 6 samples. The sequence type (ST) was identified as ST50 for both samples where the profile corresponded to L. interrogans species and Bataviae serogroup. The presence of this genotype of Leptospira has never been reported in Thailand. Thus, our findings showed the existence of ST50 L. interrogans serogroup Bataviae and the ability to cause leptospirosis in dogs in Thailand.

Spike protein D614G and RdRp P323L: the SARS-CoV-2 mutations associated with severity of COVID-19

  • Biswas, Subrata K.;Mudi, Sonchita R.
    • Genomics & Informatics
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    • v.18 no.4
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    • pp.44.1-44.7
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    • 2020
  • The severity of coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), greatly varies from patient to patient. In the present study, we explored and compared mutation profiles of SARS-CoV-2 isolated from mildly affected and severely affected COVID-19 patients in order to explore any relationship between mutation profile and disease severity. Genomic sequences of SARS-CoV-2 were downloaded from Global Initiative on Sharing Avian Influenza Data (GISAID) database. With the help of Genome Detective Coronavirus Typing Tool, genomic sequences were aligned with the Wuhan seafood market pneumonia virus reference sequence and all the mutations were identified. Distribution of mutant variants was then compared between mildly and severely affected groups. Among the numerous mutations detected, 14408C>T and 23403A>G mutations resulting in RNA-dependent RNA polymerase (RdRp) P323L and spike protein D614G mutations, respectively, were found predominantly in severely affected group (>82%) compared with mildly affected group (<46%, p < 0.001). The 241C>T mutation in the non-coding region of the genome was also found predominantly in severely affected group (p < 0.001). The 3037C>T, a silent mutation, also appeared in relatively high frequency in severely affected group compared with mildly affected group, but the difference was not statistically significant (p = 0.06). We concluded that spike protein D614G and RdRp P323L mutations in SARS-CoV-2 are associated with severity of COVID-19. Further studies will be required to explore whether these mutations have any impact on the severity of disease.

Phylogenetic ANalysis of Hepatitis G Virus by Group-Specific Sequences in the 5-Untranslated Region (5'-UTR 영역의 그룹특이적 염기서열에 의한 HGV의 계통분석)

  • Kim, Pu-Kyung;Park, Sung-Woo;Kim, Chong-Kyung;Baik, Hyung-Suk;Jang, Kyung-Lib
    • Journal of Life Science
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    • v.8 no.3
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    • pp.279-284
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    • 1998
  • The nucleotide sequences of the 5'-untraslated region(5'-UTR) of Hepatitis G virus(HGV) from sera of Korean patients were determines. When compared to the previously reported isolates, the Korean isolates have higher sequence homology with the Japanese isolates indicating the geographic distribution of HGV variants. Interestingly, three discrete regions which are highly conserved among HGV isolates from the same geographical area, thus could be applied to distinguish HGV isolates from the different areas were noticed in the 5'-UTR. Based on the sequences of these group-specific regions, twenty four different HGV isolates could be classified into 5 groups. By using the group-specific regions, inconsistency in HGV typing when based on the different regions of HGV could be solved.

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Transmission Aspect of Methicillin-resistant Staphylococcus aureus in the Neurosurgical Intensive Care Unit by Analysing Genotype (유전자형 분석에 의한 신경외과 중환자실의 메티실린 내성 황색포도알균의 전파양상)

  • Kim, Yun-Kyung;Hong, Hae-Sook
    • Journal of Korean Academy of Nursing
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    • v.37 no.6
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    • pp.976-985
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    • 2007
  • Purpose: The aim of this study was to identify the present situation of hospital infection and route of infection by clarifying the transmission aspect of methicillin-resistant Staphylococcus aureus(MRSA) in a Neurosurgical Intensive Care Unit by analysing genotype. Methods: MRSA was cultured from twenty five patients with a tracheostomy, twenty five health care workers, and environments in the Neurosurgical Intensive Care Unit of one hospital in D city. Data was collected from December 21, 2004 to November 5, 2005. MRSA isolates representing each genotype were analyzed by spaA typing and a multiplex PCR method capable of identifying the structural type of the staphylococcal cassette chromosome mec(SCCmec) carried by the bacteria. Results: As the same genotype and gene sequence were found among health care workers, patients, and environments, it was assumed that there was cross transmission among them. Conclusion: This study suggests that first, as the hospital infection by MRSA between health care workers and patients in the Neurosurgical Intensive Care Unit was due to result of cross transmission and the relevance of transmission between them was verified, it is necessary to take preventive measures and conduct education. Secondly, development of nursing interventions and study of infection are needed. Thirdly, consistent investment in prevention against hospital infections and environmental renovation is needed.