Molecular Typing of Pseudomonas aeruginosa by Randomly Amplified Polymorphic DNA

양병선 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 2003 Journal of biomedical laboratory sciences Vol.9 No.4

Pseudomonas aerugionsa is a commonly isolated nosocomial pathogen. DNA fingerprinting of P. aerugionsa is examined by randomly amplified polymorphic DNA (RAPD). In this study, P. aeruginosa were isolated from environmental and clinical specimens and the molecular typing of the microorganisms was investigated by RAPD. Thirty strains of P. aeruginosa were selected from the strains isolated formerly and submitted for type identification to the University Hospital. 15 strains of P. aeruginosa were received from Chungnam University Hospital and 14 strains from Gyeongsang University Hospital. DNA of P. aeruginosa was extracted by Qiagen genomic DNA kit. PCR mixtures were set up and incubated. Reactions mixtures were made to be optimal for P. aeruginosa. RAPD typing analysis was carried out by the multivariate statistical program (MVSP) V3.0. RAPD type I was the most common pattern and included 23 strains. Most of strains from Gyeongsang University Hospital belonged to RAPD type Ib and 15 strains from Chungnam University Hospital to RAPD type Ⅰ or Ⅱ. RAPD typing of P. aeruginosa isolated from the environmental and clinical specimens was very simple and reproducible.

중합효소연쇄반응법에 의한 메치실린내성황색포도구균의 검출

양병선 진주간호보건전문대학 2000 진주간호보건전문대학 논문집 Vol.23 No.1

Molecular Epidemiology of Listeria monocytogenes by Ribotyping

양병선 대한의생명과학회 2002 Biomedical Science Letters Vol.8 No.2

Ten Listeria monocytogenes were isolated from clinical specimens and mussels, and their physio-biochemical characters were compared with the type strains. Ribotyping was used as a taxonomic tool to determine molecular epidemiological marker. Chromosomal DNA was cleaved with restriction enzymes HindIII and EcoRI. The fragment were subjected to Southern blot hybridization with 16S rDNA from B. subtilis by PCR. EcoRI patterns of Listeria strains showed 6 to 8 bands ranging from 0.75 kb to 11 kb band and they were classified into 6 groups. In comparison, HindIII patterns revealed that 5 to 7 bands ranging from 2.75 kb to 7.75 kb band and they classified into 5 groups. The various patterns of Listeria strains were observed within genus, species and isolated sources. 16S rRNA gene restriction patterns (ribotyping) are useful in epidemiological and taxonomic study.

Molecular Epidemiology of Listeria monocytogenes by Ribotyping

Yang, Byoung-Seon 대한의생명과학회 2002 Journal of biomedical laboratory sciences Vol.8 No.2

Ten Listeria monocytogenes were isolated from clinical specimens and mussels, and their physio-biochemical characters were compared with the type strains. Ribotyping was used as a taxonomic tool to determine molecular epidemiological marker. Chromosomal DNA was cleaved with restriction enzymes HindIII and EcoRI. The fragment were subjected to Southern blot hybridization with 165 rDNA from B. subtilis by PCR. EcoRI patterns of Listeria strains showed 6 to 8 bands ranging from 0.75 kb to 11 kb band and they were classified into 6 groups. In comparison, HindIII patterns revealed that 5 to 7 bands ranging from 2.75 kb to 7.75 kb band and they classified into 5 groups. The various patterns of Listeria strains were observed within genus, species and isolated sources. 165 rRNA gene restriction patterns (ribotyping) are useful in epidemiological and taxonomic study.

Rapid Detection of Vancomycin-resistance Enterococci by SYBR Green Real-time PCR

Yang, Byoung-Seon 대한임상검사과학회 2014 대한임상검사과학회지(KJCLS) Vol.46 No.2

Vancomycin-resistant Enterococci (VRE) are a leading cause of a nosocomial infection. While seven glycopeptide resistance genotypes have been found in Enterococci, vanA and vanB are the most common resistance genotypes. Aims of this study were to detect antibiotic susceptibilities of 23 Enterococcus spp, which broke out in a university hospital by the disk diffusion test, to investigate specific genes of vanA and vanB by conventional and real-time PCR. PCR for vanA and vanB was performed on 23 Enterococci, all 23 were positive for vanA type. This study reports the validation of a simple and rapid VRE detection method that can be easily incorporated into the daily routine of a clinical laboratory. Early detection of VRE strains, including those with susceptibility to Vancomycin, is of paramount clinical importance, as it allows a rapid initiation of strict infection control practices as well as a therapeutic guidance for a confirmed infection. The real-time PCR method is a rapid technique to detect vanA in Enterococci. It is simple and reliable for the rapid characterization of VRE.

Phenotypic and Genotypic Detection of Metallo-β-Lactamase Producing Pseudomonas aeruginosa

Yang, Byoung-Seon,Hong, Keun-Seok,Jung, Seung-Bong,Kwon, Young-Hoon,Jeong, Jong-Yoon,Lee, Min-Joo,Lee, Hye-In,Park, Mi-Seon,Choi, Seung-Gu 대한임상검사과학회 2012 대한임상검사과학회지(KJCLS) Vol.44 No.2

This study was undertaken to evaluate phenotypic and genotypic methods for detection of Metallo-Beta-Lactamases (MBLs) among nosocomial Pseudomonas aeruginosa. Of the 50 P. aeruginosa isolates from clinical specimens, 20 were evaluated for carbapenem resistance and screened for MBL by double-disk synergy test and combined-disk test. Nineteen strains (95%) were found to be MBL producers among the 20 P. aeruginosa. MBL positives were further confirmed by Polymerase Chain Reaction (PCR). For the IMP and VIM types of MBLs, PCR analysis was performed on 19 of the 20, and 10 were positive for VIM MBL type. This study reports the validation of a simple and accurate MBL detection method that can be easily incorporated into the daily routine of a clinical laboratory. Early detection of MBL-carrying organisms, including those with susceptibility to carbapenems, is of paramount clinical importance, as it allows rapid initiation of strict infection control practices as well as therapeutic guidance for confirmed infection.Key Words : Hepatitis A virus (HAV), Anti-HAV, Hospital workers, Prevalence, Vaccination

Phenotypic and Genotypic Detection of Metallo-ß-Lactamase Producing Pseudomonas aeruginosa

( Byoung-seon Yang ),( Keun-seok Hong ),( Seung-bong Jung ),( Young-hoon Kwon ),( Jong-yoon Jeong ),( Min-joo Lee ),( Hye-in Lee ),( Mi-seon Park ),( Seung-gu Choi ) 대한임상검사과학회 2012 대한임상검사과학회지(KJCLS) Vol.44 No.2

This study was undertaken to evaluate phenotypic and genotypic methods for detection of Metallo-Beta-Lactamases (MBLs) among nosocomial Pseudomonas aeruginosa. Of the 50 P. aeruginosa isolates from clinical specimens, 20 were evaluated for carbapenem resistance and screened for MBL by doubledisk synergy test and combined-disk test. Nineteen strains (95%) were found to be MBL producers among the 20 P. aeruginosa. MBL positives were further confirmed by Polymerase Chain Reaction (PCR). For the IMP and VIM types of MBLs, PCR analysis was performed on 19 of the 20, and 10 were positive for VIM MBL type. This study reports the validation of a simple and accurate MBL detection method that can be easily incorporated into the daily routine of a clinical laboratory. Early detection of MBL-carrying organisms, including those with susceptibility to carbapenems, is of paramount clinical importance, as it allows rapid initiation of strict infection control practices as well as therapeutic guidance for confirmed infection.Key Words : Hepatitis A virus (HAV), Anti-HAV, Hospital workers, Prevalence, Vaccination

Molecular Typing of Pseudomonas aeruginosa by Randomly Amplified Polymorphic DNA

Byoung-Seon Yang 대한의생명과학회 2003 Biomedical Science Letters Vol.9 No.4

Pseudomonas aerugionsa is a commonly isolated nosocomial pathogen. DNA fingerprinting of P. aerugionsa is examined by randomly amplified polymorphic DNA (RAPD). In this study, P. aeruginosa were isolated from environmental and clinical specimens and the molecular typing of the microorganisms was investigated by RAPD. Thirty strains of P. aeruginosa were selected from the strains isolated formerly and submitted for type identification to the University Hospital. 15 strains of P. aeruginosa were received from Chungnam University Hospital and 14 strains from Gyeongsang University Hospital. DNA of P. aeruginosa was extracted by Qiagen genomic DNA kit. PCR mixtures were set up and incubated. Reactions mixtures were made to be optimal for P. aeruginosa. RAPD typing analysis was carried out by the multivariate statistical program (MVSP) V3.0. RAPD type Ⅰ was the most common pattern and included 23 strains. Most of strains from Gyeongsang University Hospital belonged to RAPD type Ib and 15 strains from Chungnam University Hospital to RAPD type Ⅰ or Ⅱ. RAPD typing of P. aeruginosa isolated from the environmental and clinical specimens was very simple and reproducible.

Random Amplified Polymorphic DNA Analysis for Typing Extended-Spectrum-β-Lactamase of Klebsiella pneumoniae

( Byoung Seon Yang ) 대한임상검사과학회 2005 대한임상검사과학회지(KJCLS) Vol.37 No.3

대학병원의 임상검체에서 extended-spectrum-β-lactamase (ESBL)생성 Klebsiella pneumoniae 51균주를 분리하였다. ESBL생성 K. pneumoniae 51균주는 광범위한 항생제에 내성을 보였고 대부분의 균주는 amikacin, gentamycin과 ciprofloxacin항생제에 내성을 나타내었다. 대학병원에서 분리한 51균주를 randomly amplified polymorphic DNA (RAPD)로 분석한 결과 충남대학병원 21균주와 충북대학병원에서 분리한 10균주는 Ⅰa 와 Ⅰb에 속하였고 경상대학 병원에서 분리한 20균주는 Ⅱa, Ⅱb에 속하였다. ESBL생성 K. pneumoniae 51균주는 RAPD 분석으로 4가지의 유전형으로 구분 할 수 있었고 유전적으로 다양하였다. 이상의 결과로 RAPD 분석은 유전형분석에 빠르고 단순하고 경제적인 방법임을 알 수 있었다. Fifty-one extended-spectrum-β-lactamase(ESBL) producing Klebsiella pneumoniae strains were isolated from national university hospitals. All K. pneumoniae strains showed resistance to broad-spectrum antibiotic and most of them presented resistance to amikacin, gentamicin and ciprofloxacin. The results of amplified polymorphic DNA (RAPD) pattern for randomly isolated fifty-one strains were as follows; both twenty-one strains from Chungnam National University hospital and ten strains from Chungbuk National University hospital showed RAPD type Ia and Ib. However, twenty strains isolated from Gyeongsang National University hospital belonged to RAPD type IIa and IIb. All isolates were divided into four molecular types and showed high level of genetic diversity. These results suggested that RAPD analysis provided a rapid and simple method for analysing genotypes of ESBL.

Preparation of Resistive-Type Glucose Sensor by Layer-by-Layer Technique and Their Properties

Yang Yu-Seon,Jeon Young-Min,Lee Chil-Won,Chang Won-Cheol,Kim Jong-Gyu,Gong Myoung-Seon,Choi Byoung-Koo,Joo Sang-Woo The Polymer Society of Korea 2006 Macromolecular Research Vol.14 No.2