Performance of a real-time PCR assay for the rapid identification of Mycobacterium species

왕혜영,김현정,김성현,김도균,조상래,이혜영 한국미생물학회 2015 The journal of microbiology Vol.53 No.1

Mycobacteria cause a variety of illnesses that differ in severityand public health implications. The differentiation ofMycobacterium tuberculosis (MTB) from nontuberculousmycobacteria (NTM) is of primary importance for infectioncontrol and choice of antimicrobial therapy. The diagnosisof diseases caused by NTM is difficult because NTM speciesare prevalent in the environment and because they have fastidiousproperties. In the present study, we evaluated 279clinical isolates grown in liquid culture provided by TheCatholic University of Korea, St. Vincent’s Hospital usingreal-time PCR based on mycobacterial rpoB gene sequences. The positive rate of real-time PCR assay accurately discriminated100% (195/195) and 100% (84/84) between MTB andNTM species. Comparison of isolates identified using theMolecuTech REBA Myco-ID® and Real Myco-ID® were completelyconcordant except for two samples. Two cases thatwere identified as mixed infection (M. intracellulare-M. massilienseand M. avium-M. massiliense co-infection) by PCRREBAassay were only detected using M. abscessus-specificprobes by Real Myco-ID®. Among a total of 84 cases, themost frequently identified NTM species were M. intracellulare(n=38, 45.2%), M. avium (n=18, 23.7%), M. massiliense(n=10, 13.2%), M. fortuitum (n=5, 6%), M. abscessus(n=3, 3.9%), M. gordonae (n=3, 3.9%), M. kansasii (n=2,2.4%), M. mucogenicum (n=2, 2.4%), and M. chelonae (n=1, 1.2%). Real Myco-ID® is an efficient tool for the rapid detectionof NTM species as well as MTB and sensitive andspecific and comparable to conventional methods.

Detection of Rifampicin- and Isoniazid-Resistant Mycobacterium tuberculosis Using the Quantamatrix Multiplexed Assay Platform System

왕혜영,어영,김서용,조은진,이종석,이혜영 대한진단검사의학회 2018 Annals of Laboratory Medicine Vol.38 No.6

Background: The increasing prevalence of drug-resistant tuberculosis (TB) infection represents a global public health emergency. We evaluated the usefulness of a newly developed multiplexed, bead-based bioassay (Quantamatrix Multiplexed Assay Platform [QMAP], QuantaMatrix, Seoul, Korea) to rapidly identify the Mycobacterium tuberculosis complex (MTBC) and detect rifampicin (RIF) and isoniazid (INH) resistance-associated mutations. Methods: A total of 200 clinical isolates from respiratory samples were used. Phenotypic anti-TB drug susceptibility testing (DST) results were compared with those of the QMAP system, reverse blot hybridization (REBA) MTB-MDR assay, and gene sequencing analysis. Results: Compared with the phenotypic DST results, the sensitivity and specificity of the QMAP system were 96.4% (106/110; 95% confidence interval [CI] 0.9072–0.9888) and 80.0% (72/90; 95% CI 0.7052–0.8705), respectively, for RIF resistance and 75.0% (108/144; 95% CI 0.6731–0.8139) and 96.4% (54/56; 95% CI 0.8718–0.9972), respectively, for INH resistance. The agreement rates between the QMAP system and REBA MTB-MDR assay for RIF and INH resistance detection were 97.6% (121/124; 95% CI 0.9282–0.9949) and 99.1% (109/110; 95% CI 0.9453–1.0000), respectively. Comparison between the QMAP system and gene sequencing analysis showed an overall agreement of 100% for RIF resistance (110/110; 95% CI 0.9711–1.0000) and INH resistance (124/124; 95% CI 0.9743–1.0000). Conclusions: The QMAP system may serve as a useful screening method for identifying and accurately discriminating MTBC from non-tuberculous mycobacteria, as well as determining RIF- and INH-resistant MTB strains.

TaqMan 실시간 PCR법에 의한 개 전염성 간염 바이러스의 검출

왕혜영,최재용,이미진,박진호,조매림,한재철,최경성,채준석,Wang, Hye-young,Choi, Jae-yong,Lee, Mi-jin,Park, Jin-ho,Cho, Mae-Rim,Han, Jae-cheol,Choi, Kyoung-seong,Chae, Joon-seok 대한수의학회 2004 大韓獸醫學會誌 Vol.44 No.4

The aim of this work was the validation of a rapid real-time PCR assay based on TaqMan technology for the unequivocal identification of infectious canine hepatitis (ICH) virus, to be used directly on DNA purified from blood specimens. A real-time PCR system targeting at the E3 ORFA gene sequence of canine adenovirus type 1 was optimized and validated through comparative analysis of samples using conventional PCR system. The real-time PCR assay based on TaqMan technology could disclose 23 (37.7%) out of 61 samples as PCR positive. In contrast, 18 (29.5%) samples were found PCR positive when conventional PCR was applied on these samples. The use of the ABI Prism 7700 sequence detection system allowed the efficient determination of the amplified product accumulation through a fluorogenic probe. The entire real-time TaqMan PCR assay, including DNA extraction, amplification, and detection could be completed within 3 hours. The detection method of real-time TaqMan PCR assay was 1,000 times more sensitive than conventional PCR. Real-time TaqMan probe and primer set developed and optimized in this study is a sensitive, rapid and accurate method for detection of ICH virus and can be effective screening tool for the detection of ICH in a diagnostic laboratory routines.

도말 양성 임상검체에서 MolecuTech Real MTB-ID을 이용한 결핵균과 비결핵균의 검출평가

왕혜영,이혜영,진현우,방혜은,최연임,박은미,고원중 대한임상미생물학회 2011 Annals of clinical microbiology Vol.14 No.3

Background: The differentiation of Mycobacterium tuberculosis (MTB) from nontuberculous mycobacteria (NTM) is of primary importance for infection control and choice of antimicrobial therapy. The diagnosis of diseases caused by NTM is difficult, because NTM are prevalent in the environment such as soil and water, and because they have fastidious properties. In this study we evaluated real-time PCR based MTB/NTM detection kit for its usefulness in discrimination of MTB and NTM species. Methods: A total of one hundred fifty five sputum specimens whose AFB staining smear and culture had been positives were used for this study. Among them, 59 and 96 had been identified as MTB and NTM, respectively. DNAs obtained from sputum specimens were subjected for MolecuTech Real MTB-IDⓇ (M&D, Korea), real-time PCR-based MTB/ NTM detection kit. Subsequently, the results of MolecuTech Real MTBIDⓇ were compared with AFB staining smear and culture results. Results: The positive rate of MolecuTech Real MTBIDⓇ to detect MTB and NTM were 98.3% (58/59) and 97.9 (94/96) respectively using sputum specimens. Conclusion: For detection of MTB/NTM, the sensitive and specific of MolecuTech Real MTB-IDⓇ was comparable to those of conventional methods. Therefore, this study suggests the usefulness of real-time PCR based MolecuTech Real MTB-IDⓇ for rapid and detection of MTB/NTM from direct specimens.

군산인근해역에서 분리동정된 Vibrio 속의 특성과 해수에서의 생존

왕혜영(Hye Young Wang),이건형(Geon Hyoung Lee) 한국환경생물학회 1999 환경생물 : 환경생물학회지 Vol.17 No.4

To investigate the population dynamics and survival of Genus Vibrio, population densities of aerobic saprophytic bacteria and Vibrio groups were measured 4 times in the intertidal waters of the Yellow Sea near Kunsan from November, 1997 to June, 1998. The distribution of heterotrophic bacteria during the survey periods by plate count and direct count method ranged from 1.2±0.6×10 exp (3) to 2.0±1.5×10 exp (4) CFU ml^-1 and from 6.0±4.0×10 exp (5) to 1.9±1.5×10 exp (7) cells ml^-1, respectively. Vibrio groups were distributed in the range of 1×10 and 6±2.2×10 exp (2) CFU ml^-1. The proportion of Vibrio groups to total heterotrophic bacteria was between 0.1 and 6% during the survey periods. A total of 51 isolates was obtained from TCBS agar plates and identified to species level by Biolog Identification System^™. As a result, dominant genera were V. mediterranei, V. anguillarum, V. metschnikovii, and V. parahaemolyticus, and isolates were clustered into 26 groups based on the relatedness of average linkage clustering method at 70% level. As for the susceptibility of 51 isolates to 7 kinds of antibacterial agents (gentamicin, ampicillin, chloramphenicol, streptomycin, kanamycin, tetracycline, carbenicillin), 96% of isolates showed high resistance to more than one antibiotics and 65% of isolates contained a plasmid, of which size was observed greater than 12kb. The number of cells of 3 tested strains (V. anguillarum, V. vulnificus, and V. metschnikovii) in filtered aged seawater decreased by approximately 1 to 5 orders of magnitude during 30-d incubation. In most cases, the numbers of cells decreased rapidly until day 3,then decreased slowly by day 30. The number of cells incubated at 15℃ showed higher survival than those at 4℃ and 25℃. These results may be considered for the basic supporting data in the risk assessment of vibriosis in summer. [Vibrio, distribution, Biolog Identification system, resistance to antibiotics, survival in seawater].

귀신문화에 기반한 일본풍의 캐릭터 디자인 분석 및 캐릭터 제작 연구 - 중국 모바일 게임 <음양사>사례를 중심으로

왕혜영(Hui-Ling Wang),이동엽(Dong-Yeop Lee) 한국게임학회 2022 한국게임학회 논문지 Vol.22 No.1

본 논문은 귀신문화에 기반을 둔 일본풍 캐릭터 디자인을 게슈탈트 시지각 법칙을 사용하여 분석한 연구이다. 중국에서 성공한 일본풍 모바일 게임 〈음양사〉의 식신(Shikigami) 캐릭터의 디자인 요소를 정리하고 분석하였다. 형태는 부분이 아니라 전체적인 체계화를 통해 지각되며 작은 디자인적 요소가 전체 스토리에 영향을 미치는 방식을 확인하였다. 캐릭터의 디자인적 요소가 시각화 되는지를 분석하고 스토리에 관련성이 있다는 것을 확인 하였다. 디자인적 요소가 이야기에 영향을 미친다는 결과를 통해 새로운 캐릭터 디자인을 제안하고 이를 실증 분석하였다. According to Gestalt theories of perception, this paper studies Japanese-style character design combined with ghost culture. This article sorts out the character design elements of the Japanese mobile game "Onmyoji" in China, and then analyzes the design from the perspective of psychology through Gestalt theories. The form is felt through the systematization of the whole rather than the parts, confirming whether the small design elements of the character can be perceived as the whole story.

Clinical Characteristics and Genetic Variations in Early-Onset Atopic Dermatitis Patients

김범준,왕혜영,이혜영,이소연,최응호,홍수종 대한피부과학회 2019 Annals of Dermatology Vol.31 No.3

Background: Hereditary factors contribute to atopic dermatitis (AD) development. We developed the reverse blot hybridization assay (REBA) kit to simultaneously detect variations in skin barrier- and immune response-related genes prevalent in Korean AD patients. Objective: To identify genetic variations and clinical characteristics that could predict early AD development. Methods: We compared AD-related genetic variations between early-onset AD subjects and non-AD controls, and clinical characteristics and genetic variations between early- and late-onset AD subjects. We compared 28 early-onset AD subjects and 57 non-AD controls from a birth cohort and 108 early- (age ≤3 years) and 90 late-onset AD subjects and 189 non-AD controls from a university hospital. Genetic variations were detected via REBA. Results: There were no differences in AD-related genetic variation between early-onset AD subjects and non-AD controls in the birth cohort. When the birth cohort and hospital populations were combined, early-onset AD subjects and non-AD controls showed different frequencies of genetic variations of KLK7, SPINK5 1156, DEFB1, IL5RA, IL12RB1a, and IL12RB1b. No differences in the frequency of genetic variations were observed between early- and late-onset AD subjects. Immunoglobulin E positivity for house dust mites was prevalent in late-onset AD subjects. A family history of atopic diseases was associated with early-onset AD. Conclusion: No AD-related genetic variations could predict early AD development in Koreans, even though neonates with a family history of atopic diseases are likely to develop AD at ≤3 years of age. Environmental exposure may be more important than genetic variation in determining the onset age of AD.

Differences in Genetic Variations Between Treatable and Recalcitrant Atopic Dermatitis in Korean

전명수,왕혜영,이소람,최은희,이혜영,최응호 대한천식알레르기학회 2018 Allergy, Asthma & Immunology Research Vol.10 No.3

Purpose: Variations in barrier- or immune response-related genes are closely related to the development of atopic dermatitis (AD). This study was designed to identify genetic variations and clinical features to predict ‘recalcitrant AD.’ Methods: AD patients were classified as treatable and recalcitrant. Treatable AD patients showed satisfactory clinical improvement with basic and topical treatments. Recalcitrant AD patients used systemic immune-suppressants for over 4 weeks as they had not shown clinical improvement with basic and topical treatments. The frequency of gene variations in barrier- (FLG 3321delA, FLG K4022X, KLK7, SPINK 1156, SPINK 1188, SPINK 2475) and immune response- (DEFB1, KDR, IL-5RA, IL-9, and IL-12RB1a, b) related genes were compared between each AD group and the controls. Results: Of all, 249 treatable AD and 32 recalcitrant AD were identified. Heterozygous mutations (Hetero) in KLK7 was more frequent in recalcitrant AD patients than treatable AD, without statistical significance. Hetero in DEFB1 was more frequent in treatable AD patients. However, no other significant genetic differences between treatable and recalcitrant AD was observed. Instead, higher initial Eczema Area Severity Index (EASI) score, serum immunoglobulin E (IgE) level, allergen specific IgE for house dust mites, and family history of atopic diseases were associated with recalcitrant AD with statistical significance. Conclusions: According to our study, no genetic variation to predict recalcitrant AD was identified, suggesting that clinical manifestation, rather than genetic variations of AD patients is more likely to be an important factor in predicting the prognosis of AD. Further large-scale studies on the correlation between genetic variation and recalcitrant AD are needed.

NDM-5 and OXA-48 Co-producing Uropathogenic Escherichia coli Isolate: First Case in Korea

장진호,왕혜영,유길성,황규열,어영,윤갑준 대한진단검사의학회 2018 Annals of Laboratory Medicine Vol.38 No.3

Chungang University

이혜영,왕혜영 대한의생명과학회 2007 Biomedical Science Letters Vol.13 No.1