G+C 함량이 높은 Primer를 사용하는 중합효소 연쇄반응에서 변성제가 미치는 영향

김영미,안준환,김종배,엄용빈 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1996 Journal of biomedical laboratory sciences Vol.2 No.2

G+C 함량이 높은 primer를 이용한 중합효소 연쇄반응을 실시하는 경우 높은 annealing 온도로 인하여 특이 염기서열의 합성정도가 매우 미약하게 나타나는 경우가 많이 있다. 이와 같은 문제점을 보완하기 위하여 glycerol, formamide 및 dimethyl sulfoxide (DMSO) 등의 변성제를 반응용 완충액에 첨가하고 중합효소 연쇄반응을 실시하여 그 결과를 비교 검토하였다. G+C 함량이 낮은 Borrelia burgdorferi의 Ly1 유전자의 primer set인 Bb 679와 Bb 680를 이용한 중합효소 연쇄반응에서는 변성제 첨가에 따른 합성 DNA의 양의 변화가 뚜렷하지 않았다. 그러나 G+C 함량이 높은 primer set인 Mycobacterium paratuberculosis의 IS900 유전자의 IS900/150C와 IS900/921를 이용한 중합효소 연쇄반응을 유도한 경우에는 변성제를 첨가함에 따라 합성된 DNA의 양의 증가가 뚜렷하였으며, 2.5% glycerol과 1.25% formamide를 혼합 첨가한 경우와 또는 2.5% DMSO를 반응용 완충액에 첨가하였을 때 비특이적인 증폭비율이 낮아 특이 염기서열의 합성 결과가 양호한 것으로 나타났다. Poor yields of amplified DNAs could be resulted in polymerase chain reaction(PCR) processes with G+C-rich DNA primers because of their high Tm values. To maximize the yields of amplification in PCR processes with G+C-rich primers, we compared the yields of amplified DNA fragments according to the concentrations of specific denaturants added to the reaction mixture of PCR system. With addition of the mixture of 2.5% glycerol and 1.25% formamide, or 2.5% dimethyl sulfoxide to the reaction cocktail, respectively, remarkable increases in the yields of amplified DNA fragments were not observed in the PCR systems with G+C-low primers of Lyl chromosomal gene from Borrelia burgdorferi but observed in the PCR system with G+C­rich primers of IS900 gene from Mycobacterium paratuberculosis. Although we were not practically able to discriminate the yields of PCR DNAs according to the concentrations used in this study, addition of the mixture of 5% glycerol and 2.5% formamide, or 5% DMSO tended to increase the production of extra bands.

개인식별을 위한 법의 유전자 분석

엄용빈 순천향대학교 기초과학연구소 2014 순천향자연과학연구 논문집 Vol.20 No.1

Since DNA fingerprinting method was introduced in 1985, human DNA identification has steadily worked its way into the routine of criminal investigation and has now become an indispensable tool. Forensic DNA analysis has been applied in various aspects of the identification of persons, the conviction or exoneration of suspects and the identification of victims of crimes, accidents and disasters. Autosomal short tandem repeat (STR) profiles are generated from biological materials found at crime scenes and compared with profiles of known suspects identified by police investigations or included in national forensic DNA databases. A STR profile match provides strong evidence for individual identification. The use of STR profiling to identify perpetrators, disaster victims and family members has proven to be very successful. We now address how novel forensic DNA typing achieve high levels of individualization and provide investigators with authentic links between DNA-identified sample donors and specific criminal acts. This review aims to discuss how recent advances in forensic DNA profiling fields that include the improved ability to analyse degraded DNA and low amounts of DNA, an increase in discrimination power, and the application of STR profiling for familial searching.

Multiplex PCR을 이용한 장출혈성 대장균 0157:H7의 검출

김종배,엄용빈 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1998 Journal of biomedical laboratory sciences Vol.4 No.1

최근 전세계적으로 문제가 되고 있는 장출혈성 대장균 0157:H7을 분리배양 및 동정없이 바로 시료를 분석하여 신속하게 검출하기 위한 다중 중합효소 연쇄반응 (multiplex PCR) 기법을 확립하고, 이 기법을 이용하여 국내 분리 균주 중에서 SLT-I·Ⅱ, eaeA, 60-MDa plasmid gene을 가지고 있는 대장균을 유전자 수준에서 검출하고자 하였다. 장출혈성 대장균 0157:H7이 가진 SLT-I·Ⅱ, eaeA, 60-MDa plasmid 유전자들에 대한 특이 oligonucleotide primers (MK1'-MK2', NAE19-NAE20, MFS1F-MFS1R)를 함께 동시에 반응 완충액에 넣어 다중 중합효소 연쇄반응을 시행한 결과 317bp (eaeA), 228bp (SLT-I·Ⅱ), 167bp (60-MDa plasmid)의 PCR 증폭 DNA 생성물을 표준균주 (E. coli ATCC 35150)에서는 확인할 수 있었지만, 기타 다른 병원성 장내세균 13균주에서는 band를 확인할 수 없었다. 한편 다중 중합효소 연쇄반응의 template DNA 추출 방법에 따른 PCR 결과를 비교하였다. 각각의 DNA 추출 방법 중 boiling lysis 방법이 신속하고 간편하여 장출혈성 대장균 0157:H7에 의한 식중독의 임상진단에 다중 중합효소 연쇄반응 (multiplex PCR) 적용하는 데에는 boiling lysis법을 이용하는 것이 가장 적합한 방법으로 확인되었다. A multiplex PCR method was designed by employing primers specific for the eaeA gene, conserved sequences of Shiga-like toxins (SLT-I·Ⅱ), and the 60-MDa plasmid of enterohemorrhagic E. coli (EHEC) 0157:H7 strain. A set of six synthetic oligonucleotide primers derived from sequences of the SLT-I·Ⅱ, eaeA, and 60-MDa plasmid genes of E. coli 0157:H7 were used in a multiplex PCR amplification procedure to detect these genes in the same enteric pathogens. In two enterohemorrhagic E. coli 0157:H7 (ATCC 35150, ATCC 43894) reference strains, PCR products of 317bps (eaeA), 228bps (SLT-I·Ⅱ) and 167bps (60-MDa plasmid) were successfully amplified simultaneously in a single reaction. However, the specific PCR products were not amplified in control strains of other enteric bacteria. The sensitivity of the multiplex PCR assay for detection of the SLT-I·Ⅱ, eaeA, and 60-MDa plasmid genes of E. coli 0157:H7 was found to be 2.5×10 of bacteria in diarrheal stool to amplify all three bands. The multiplex PCR technology will allow large-scale screening of many clinical specimens or contaminated foods, and will be a very useful method for the detection of a wide range of microorganisms present in the environment, including EHEC 0157:H7 in various types of specimens. The multiplex PCR assay has the potential to be used as a specific and rapid method for clinical diagnosis of disease caused by EHEC 0157:H7.

임상병리학 교육의 수월성 강화를 위한 교과과정의 질적 개선에 대한 비교 연구

엄용빈 나사렛대학교출판사 2011 지성과 창조 Vol.- No.14

Multiplex PCR을 이용한 장출혈성 대장균 O157:H7의 검출

엄용빈,김종배 대한의생명과학회 1998 Journal of biomedical laboratory sciences Vol.4 No.1

최근 전세계적으로 문제가 되고 있는 장출형성 대장균 O157:H7을 분리배양 및 동정 없이 바로 시료를 분석하여 신속하게 검출하기 위한 다중 중합효소 연쇄반응 (multiplex PCR) 기법을 확립하고, 이 기법을 이용하여 국내 분리 균주 중에서 SLT-I.II, eaeA, 60-MDa plasmid gene을 가지고 있는 대장균을 유전자 수준에서 검출하고자 하였다. 장출혈성 대장균 O157:H7이 가진 SLT-I.II, 60-MDa plasmid 유전자들에 대한 특이 oligonucleotide primers (MK1'-MK2', NAE19-NAE20, MFSIF-MFSIR)를 함께 동시에 반응 완충액에 넣어 다중 중합효소 연쇄반응을 시행한 결과 317bp (eaeA), 228bp (SLT-I.II), 167bp (60-MDa plasmid)의 PCR 증폭 DNA생성물을 표준균주 (E. coli ATCC 35150)에서는 확인할 수 있었지만, 기타 다른 병원성 장내세균 13세균 13균주에서는 band를 확인할 수 없었다. 한편 다중 중합효소 연쇄반응의 template DNA 추출 방법에 따른 PCR 결과를 비교하였다. 각각의 DNA 추출 방법 중 boiling lysis 방법이 신속하고 간편하여 장출혈성 대장균 O157:H7에 의한 식중독의 임상진단에 다중 중합효소 연쇄반응 (multiplex PCR) 적용하는 데에는 boiling lysis법을 이용하는 것이 가장 적합한 방법으로 확인되었다. A multiplex PCR method was designed by employing primers specific for the eaeA gene, conserved sequences of Shiga-like toxins (SLT-I.II), and the 60-MDa plasmid of enterohemorrhagic E. coli (EHEC) O157:H7 strain. A set of six synthetic oligonucleotide primers derived from sequences of the SLT-I.II, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 were used in a multiplex PCR amplification procedure to detect these genes in the same enteric pathogens. In two enterohemorrhagic E. coli O157:H7 (ATCC 35150, ATCC 43894) reference strains, PCR products of 317bps (eaeA), 228bps (SLT-I.II), and 167bps (60-MDa plasmid) were successfully amplified simultaneously in a single reaction. However, the specific PCR products were not amplified in control strains of other enteric bacteria. The sensitivity of the multiplex PCR assay for detection of the SLT-I.II, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 was found to be 2.5$\times$10$^{6}$ of bacteria in diarrheal stool to amplify all three bands. The multiplex PCR technology will allow large-scale screening of many clinical specimens or contaminated foods, and will be a very useful method for the detection of a wide range of microorganisms present in the environment, including EHEC O157:H7 in various types of specimens. The multiplex PCR assay has the potential to be used as a specific and rapid method for clinical diagnosis of disease caused by EHEC O157:H7.

정보통신공사 하자관리 실태조사를 통한 세부 공종별 하자 유형 및 원인 분석

엄용빈 ( Eom Yong-been ),손성근 ( Sohn Sung-geun ),강상훈 ( Kang Sang-hun ),박현정 ( Park Hyun Jung ) 한국건축시공학회 2021 한국건축시공학회 학술발표대회 논문집 Vol.21 No.1

As the era of the Fourth Industrial Revolution arrives and the culture of contact-less was established due to the long-term COVID-19, the construction industry is striving to develop smart home services to expand the scope of information and communications technology (ICT) to provide convenience to residents. However, various defects are occurring in information and communications technology facilities in apartments, and related research is lacking. In this study, case studies and research methods of related prior studies are analyzed as a basis for identifying the types and causes of defects that occur in information and communications technology facilities. To analyze various studies, we divided them into studies on smart home Internet of Things (IoT), information and communications technology construction of apartments, and investigated their features and limitations. Plus, future research will be conducted on apartments within 10 years of completion among apartments completed by domestic construction companies to supplement the limitations of prior research. In addition, we will prepare basic data on defects of ICT construction by analyzing the major causes of defect types through the analysis of defect types and frequency by detailed work type of information and communications technology construction.

Multiplex PCR을 이용한 장출혈성 대장균 O157:H7의 검출

엄용빈(Yong-Bin Eom),김종배(Jong-Bae Kim) 대한의생명과학회 1998 Biomedical Science Letters Vol.4 No.1

최근 전세계적으로 문제가 되고 있는 장출혈성 대장균 O157:H7을 분리배양 및 동정없이 바로 시료를 분석하여 신속하게 검출하기 위한 다중 중합효소 연쇄반응 (multiplex PCR) 기법을 확립하고, 이 기법을 이용하여 국내 분리 균주 중에서 SLT-ⅠㆍⅡ, eaeA, 60-MDa plasmid gene을 가지고 있는 대장균을 유전자 수준에서 검출하고자 하였다. 장출혈성 대장균 O157:H7이 가진 SLT-ⅠㆍⅡ, eaeA, 60-MDa plasmid 유전자들에 대한 특이 oligonucleotide primers (MKI'-MK2', NAEI9-NAE20, MFSIF-MFSIR)를 함께 동시에 반응 완충액에 넣어 다중 중합효소 연쇄반응을 시행한 결과 317bp (eaeA), 228bp (SLT-ⅠㆍⅡ), 167bp (60-MDa plasmid)의 PCR 증폭 DNA생성물을 표준균주 (E. coli ATCC 35150)에서는 확인할 수 있었지만, 기타 다른 병원성 장내세균 13균주에서는 band를 확인할 수 없었다. 한편 다중 중합효소 연쇄반응의 template DNA 추출 방법에 따른 PCR 결과를 비교하였다. 각각의 DNA 추출 방법 중 boiling lysis 방법이 신속하고 간편하여 장출혈성 대장균 O157:H7에 의한 식중독의 임상진단에 다중 중합효소 연쇄반응 (multiplex PCR) 적용하는 데에는 boiling lysis법을 이용하는 것이 가장 적합한 방법으로 확인되었다. A multiplex PCR method was designed by employing primers specific for the eaeA gene, conserved sequences of Shiga-like toxins (SLT-ⅠㆍⅡ), and the 60-MDa plasmid of enterohemorrhagic E. coli (EHEC) 0157:H7 strain. A set of six synthetic oligonucleotide primers derived from sequences of the SLT-ⅠㆍⅡ, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 were used in a multiplex PCR amplification procedure to detect these genes in the same enteric pathogens. In two enterohemorrhagic E. coli O157:H7 (ATCC 35150, ATCC 43894) reference strains, PCR products of 317bps (eaeA), 228bps (SLT-ⅠㆍⅡ), and 167bps (60-MDa plasmid) were successfully amplified simultaneously in a single reaction. However, the specific PCR products were not amplified in control strains of other enteric bacteria. The sensitivity of the multiplex PCR assay for detection of the SLT-ⅠㆍⅡ, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 was found to be 2.5×10? of bacteria in diarrheal stool to amplify all three bands. The multiplex PCR technology will allow large-scale screening of many clinical specimens or contaminated foods, and will be a very useful method for the detection of a wide range of microorganisms present in the environment, including EHEC O157:H7 in various types of specimens. The multiplex PCR assay has the potential to be used as a specific and rapid method for clinical diagnosis of disease caused by EHEC O157:H7.

원저(原著) : 소아 수혈용 소단위 적혈구 농축액 제조 및 사용경험

엄용빈 ( Yong Bin Eom ),배인철 ( In Cheol Bse ),안효준 ( Hyo Jun Ahn ),이정신 ( Jung Sin Lee ),김현옥 ( Hyun Ok Kim ) 대한임상병리사협회 1996 임상수혈검사학회 발표자료집 Vol.3 No.1

For pediatric and neonatal transfusions, it is important to supply the volume needed. Newborns, compared to adult, require transfusion more frequently, which increases donor exposure and donor-related risks such as hepatitis. The unique small-volume requirements of transfusion to neonates may give rise to a large amount of wastage. Thus, a single donor unit for adult transfusion is not adequate for the pediatric and neonatal transfusion. Our technical approaches for pediatric units are available to supply the small volume needed and decrease blood wastage associated with requests for small volume transfusion and reduce donor exposure for those infants who are maintained with transfusion from the same unit for several days (14 days). Small-volume bags is attached to tubing from the outlet port of the main bag using the Sterile Connective Device. The repeated manipulation needed to make small-volume aliquots of packed RBCs did not result in contamination. This method for pediatric transfusion was simple for preparation and easy to adjust the volume. The outcome of the transfused patients was successful. In Korea, transfusion services providing blood for pediatrics and neonates need to develop the methods of preparing 70~120 ml pediatric units of whole blood or red blood cells to realize above advantages and to minimize wastage.

부검조직의 부패정도와 면역조직화학염색상 염색도의 비교 고찰

김영주,엄용빈,권태정,김유희 내무부 국립과학수사연구소 2000 國立科學搜査硏究所年報 Vol.32 No.-

R1R1(DCE/DEc) 환자에서 항-E항체와 항-C 항체의 동시 감작에 관한 고찰

안효준,엄용빈,이정신,최민자,김현옥 大韓輸血學會 1996 大韓輸血學會誌 Vol.7 No.2