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 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

O-5 : Detection of SHV and CTX-M Extended-Spectrum β-Lactamases Producing Gram-Negative Rods by Multiplex Real-Time PCR

( Byoung Seon Yang ),( Keun Dol Yook ) 대한임상병리사협회 2009 임상미생물검사학회 발표자료집 Vol.2009 No.-

Background: The production of extended-spectrum β-lactamases (ESBLs) by bacterial pathogens is a major threat to the use of the clinically important expanded-spectrum cephalosporins. In this study, we demonstrated a rapid, sensitive, and specific method for the detection of SHV and CTX-M ESBLs producing gram-negative bacilli using a multiplex PCR that uses SYBR Green labeled on a Smart Cycler instrument. Methods: Forty Gram-negative bacilli were isolated from the university hospital in Daejeon and an antibiotic susceptibility test was performed using CLSI guidelines. The confirmation of ESBL producing Gram-negative bacilli was performed by double disk synergy test. The DNA used for PCR was isolated by using the Bioneer Plasmid mini extraction kit. Multiplex Real-Time PCR analysis was performed using SYBR Premix ExTaq reagents and its technology. Results: Forty presumptive ESBL producing gram-negative bacilli were isolated according to CLSI guidelines. The double disk synergy test showed 85.5% positive ESBL producing gram-negative bacilli. SHV ESBL producing gram-negative bacilli were found in 7 of 40 (18%) isolates. CTX-M ESBL producing gram-negative bacilli were found in 9 of 40 (20%) isolates. Discussion: Detection of ESBLs in clinical isolates is difficult. The phenotypic methods are the only screening methods for the detection of ESBLs in a routine laboratory and genotypic methods help us to confirm the genes responsible for ESBL production. In conclusion, we used a multiplex real-time PCR for detection of SHV and CTX-M ESBL producing gram-negative bacilli. This method provided an efficient and rapid differentiation of ESBL producing Gram-negative bacilli and could be used for epidemiological studies among ESBL isolates.

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

Rapid Detection of Vancomycin-resistant Enterococci (VRE) in Clinical Samples from University Hospital

Yang, Byoung-Seon,Park, Jung-Yeon,Choi, Seung-Gu 대한임상검사과학회 2013 대한임상검사과학회지(KJCLS) Vol.45 No.1

Outbreaks of vancomycin-resistant enterococci (VRE) are being reported more frequently in many countries. While seven glycopeptide resistance genotypes have been described in Enterococci, vanA and vanB are the most common resistance genotypes. The aim of this study was to detect antibiotic susceptibilities of 23 Enterococcus faecium strains, which caused an outbreak in a University hospital by a disk diffusion test to investigate the presence of the species specific gene, and the resistant genotypes, vanA and vanB by duplex PCR. PCR for vanA and vanB was performed on 23 enterococci. Twenty three were identified as E. faecium and were tested positive for the vanA genotype. This study will report on the validation of a simple and accurate 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 rapid initiation of strict infection control practices, as well as the therapeutic guidance for confirmed infections. The PCR method developed in the present study is simple and reliable for the rapid characterization of VRE.

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

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.

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.

Epidemiological Investigation of Methicillin-Resistant Staphylococcus aureus by Arbitrarily Primed PCR

Yang Byoung-Seon 대한의생명과학회 2004 Biomedical Science Letters Vol.10 No.4

Methicillin-Resistant Staphylococcus aureus (MRSA) strains are resistant to a wide range of antibiotics and are a major cause of nosocomial infections. Accurate and rapid typing of MRSA is needed to implement effective infection control measures. Arbitrarily Primed PCR (AP-PCR) is a very useful method in rapid typing. AP-PCR is not necessary information about target DNA sequence because this is basically DNA amplification and could be useful in epidemiological typing by classified band pattern. In this study, MRSA were isolated and identified from ICU, Neu, IM and Ped environments and investigated molecular typing by AP-PCR. Ped, the MRSA pattern determines the la, IIa type, 1M is Ib type, Neu is IIa type and ICU determines the IIa, lIb types. All MRSA in this study were typeable by AP-PCR, which was easy to perform and reproduce with evidence of MRSA for purposes of nosocomial infection control.