• 제목/요약/키워드: multiplex-PCR

검색결과 491건 처리시간 0.029초

Virulence gene profiles and antimicrobial susceptibility of Salmonella Brancaster from chicken

  • Evie Khoo ;Roseliza Roslee ;Zunita Zakaria;Nur Indah Ahmad
    • Journal of Veterinary Science
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    • 제24권6호
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    • pp.82.1-82.12
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    • 2023
  • Background: The current conventional serotyping based on antigen-antisera agglutination could not provide a better understanding of the potential pathogenicity of Salmonella enterica subsp. enterica serovar Brancaster. Surveillance data from Malaysian poultry farms indicated an increase in its presence over the years. Objective: This study aims to investigate the virulence determinants and antimicrobial resistance in S. Brancaster isolated from chickens in Malaysia. Methods: One hundred strains of archived S. Brancaster isolated from chicken cloacal swabs and raw chicken meat from 2017 to 2022 were studied. Two sets of multiplex polymerase chain reaction (PCR) were conducted to identify eight virulence genes associated with pathogenicity in Salmonella (invasion protein gene [invA], Salmonella invasion protein gene [sipB], Salmonella-induced filament gene [sifA], cytolethal-distending toxin B gene [cdtB], Salmonella iron transporter gene [sitC], Salmonella pathogenicity islands gene [spiA], Salmonella plasmid virulence gene [spvB], and inositol phosphate phosphatase gene [sopB]). Antimicrobial susceptibility assessment was conducted by disc diffusion method on nine selected antibiotics for the S. Brancaster isolates. S. Brancaster, with the phenotypic ACSSuT-resistance pattern (ampicillin, chloramphenicol, streptomycin, sulphonamides, and tetracycline), was subjected to PCR to detect the corresponding resistance gene(s). Results: Virulence genes detected in S. Brancaster in this study were invA, sitC, spiA, sipB, sopB, sifA, cdtB, and spvB. A total of 36 antibiogram patterns of S. Brancaster with a high level of multidrug resistance were observed, with ampicillin exhibiting the highest resistance. Over a third of the isolates displayed ACSSuT-resistance, and seven resistance genes (β-lactamase temoneira [blaTEM], florfenicol/chloramphenicol resistance gene [floR], streptomycin resistance gene [strA], aminoglycoside nucleotidyltransferase gene [ant(3")-Ia], sulfonamides resistance gene [sul-1, sul-2], and tetracycline resistance gene [tetA]) were detected. Conclusion: Multidrug-resistant S. Brancaster from chickens harbored an array of virulence-associated genes similar to other clinically significant and invasive non-typhoidal Salmonella serovars, placing it as another significant foodborne zoonosis.

천안지역 소아 백일해 감염의 유병률과 임상적 고찰 (Prevalence and Clinical Characteristics of Pertussis in Children, Cheonan, Korea)

  • 이건송;손재성;정은희;홍기배;이미정;유지숙;장영표;박우성;김재경;임인수;노의정
    • Pediatric Infection and Vaccine
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    • 제16권2호
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    • pp.175-182
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    • 2009
  • 목 적: 최근 10년 동안 여러 나라에서 백일해의 발생률이 증가하고 있으며, 특히 사춘기 및 성인 연령에서 발생률이 증가하고 있음을 보고하였다. 본 연구는 천안 지역에서 백일해의 유병률과 임상양상을 알아보고 우리나라 백일해에 대한 역학 조사에 도움이 되고자 하였다. 방 법: 2008년 3월에서 2009년 9월까지 단국대학교병원 소아청소년과에 호흡기 증상으로 입원치료를 하였거나 외래 진료를 받고 호흡기 감염균 다중 PCR 검사를 시행한 118명의 환아를 대상으로 하였다. 검체는 환아의 비인두액을 무균의 카테터를 이용하여 채취하였고 이를 다중 PCR 검사를 시행하여 B. pertussis 균주의 밴드유무를 판독하고 DNA sequencing을 시행하였다. 결 과: 118명의 환아 중 10명(8%)에서 B. pertussis 양성을 보였고, 2009년 7-9월에 6명이 발생하였다. 10명 중 9명 이 3개월 미만의 영아였으며, 7명은 DTaP 예방접종을 시행하지 않은 환아였다. 진단 전 기침의 평균 기간은 10.9${\pm}$5.2일이였다. 임상양상은 발작적 기침이 10명(100%)에서 있었으며 기침 후 구토증상이 8명(80%), 기침 시 입술 주위 청색증을 보인 환아가 7명(70%) 이였다. 발열은 1명에서만 관찰되었다. 전형적인 기침 양상인 whooping은 4명(40%)에서 관찰되었다. 평균 백혈구 수는 20,068${\pm}{\pm}10,135/mm^3$ (9,280-38,320/$mm^3$) 이었고, 평균 림프구는 67.1${\pm}{\pm}$21.7% (13.3-82.5%)였다. 폐렴, 무기폐, 기종격동의 합병증과 함께 호흡곤란이 있었던 1명의 환아는 치료 중 84,400/$mm^3$의 백혈구 상승이 있었다. DTaP를 접종하지 않은 7명의 환아 중 6명에서 합병증이 발생했으며 백신 접종의 유무와 합병증 발생과는 통계적으로 유의한 연관성이 있었다(P =0.033). 결 론: 중부지역에서 백일해의 발생이 2009년 7-9월에 집중되어 있었다. 향후 소아청소년과 혹은 전국적인 감시체계 등을 통하여 국내에서의 백일해 역학의 변화에 대한 연구가 필요할 것으로 보인다.

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계절 인플루엔자 진단에서 신속항원검사의 유용성 (The Efficacy of Rapid Antigen Tests for Detection of Seasonal Influenza Virus)

  • 이준열;이솔;김한성;김광남
    • Pediatric Infection and Vaccine
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    • 제24권1호
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    • pp.31-36
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    • 2017
  • 목적: 본 연구에서는 인플루엔자 신속항원검사의 유용성을 확인해보고, 결과의 정확도에 영향을 주는 인자를 알아보고자 한다. 방법: 2011년 6월 1일부터 2016년 5월 31일까지 5년간 한림대학교 성심병원의 소아청소년과 외래와 응급실을 통해 내원한 인플루엔자 의사환자를 대상으로 하였다. 이들 중 입원하여 PCR검사를 같이 진행한 798검체의 검사 결과를 비교하였다. 결과: PCR검사를 표준으로 신속항원검사의 양성 일치율은 A형 인플루엔자에서 75.7%, B형 인플루엔자에서 60.0%였다. 발열 시작일로부터 4일 이내에 내원하여 검사를 진행한 경우 양성 일치율은 A형 인플루엔자 77.6%, B형 인플루엔자 73.2%였으나, 5일 이상 경과 후 검사를 진행한 경우 양성 일치율은 A형 인플루엔자 66.7%, B형 인플루엔자 21.4%였다. 결론: 신속항원검사의 민감도는 상대적으로 낮으므로, 인플루엔자 진단에 적용하기 위해서 증상 발현 후 5일 이내 빠르게 채취한 검체에서 상대적으로 높은 민감도 해석이 가능하다.

The Uneven Distribution of Mating Type Genes in Natural and Cultivated Truffle Orchards Contributes to the Fructification of Tuber indicum

  • Li, Qiang;Fu, Yu;Sun, Qun;Sourzat, Pierre;Yang, Mei;Liu, Chengyi;Tan, Hao;Ye, Lei;Zou, Jie;Wu, Chenguang;Zhang, Bo;Li, Xiaolin
    • Mycobiology
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    • 제46권1호
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    • pp.64-71
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    • 2018
  • The aim of this study was to investigate the pattern of distribution of mating type (MAT) genes of Tuber indicum in ectomycorhizosphere soils from natural T. indicum-producing areas and cultivated truffle orchards and ascocarp samples from different regions. Quantitative real-time PCR and multiplex PCR were used to weight the copy numbers of MAT1-1-1 and MAT1-2-1 in natural truffle soils and cultivated orchard soils. The effect of limestone on the pattern of truffle MAT genes and the correlation between soil properties and the proportion of MAT genes were also assessed. These results indicated that an uneven and nonrandom distribution of MAT genes was common in truffle-producing areas, cultivated truffle orchards, and ascocarps gleba. The competition between the two mating type genes and the expansion of unbalanced distribution was found to be closely related to truffle fructification. Limestone treatments failed to alter the proportion of the two mating type genes in the soil. The content of available phosphorus in soil was significantly correlated with the value of MAT1-1-1/MAT1-2-1 in cultivated and natural ectomycorhizosphere soils. The application of real-time quantitative PCR can provide reference for monitoring the dynamic changes of mating type genes in soil. This study investigates the distributional pattern of T. indicum MAT genes in the ectomycorhizosphere soil and ascocarp gleba from different regions, which may provide a foundation for the cultivation of T. indicum.

Molecular Typing and Antimicrobial Susceptibility of Staphylococcus aureus Strains Isolated from Raw Milk, Cheese, Minced Meat, and Chicken Meat Samples

  • Can, Hayriye Yesim;Elmali, Mehmet;Karagoz, Alper
    • 한국축산식품학회지
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    • 제37권2호
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    • pp.175-180
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    • 2017
  • The objectives of this study were: i) to detect the presence of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) in raw milk, cheese, beef minced meat, and chicken meat samples; ii) to evaluate the antimicrobial susceptibility of the isolates; and iii) to determine clonal relation among the isolates by using pulsed-field gel electrophoresis (PFGE) method. Therefore, a total of 160 food samples were randomly collected between August 2014 and May 2015 in Hatay province, located in the southern Turkey. Twenty (12.5%) of the samples were found to be contaminated with S. aureus. A total of 40 isolates from the 20 positive samples were confirmed to be S. aureus by multiplex PCR based on 16S rRNA and nuc gene. The mec A gene was not detected in any of the S. aureus strains. In the present study, 39 out of 40 (97.5%) isolates were found to be resistant to one or more antibiotics. All of isolates were susceptible to gentamicin, oxacillin, and vancomycin. The highest resistance rate was detected in penicillin (95%) and ampicillin (92.5%), followed by tetracycline (30%), erythromycin (20%), ciprofloxacin (12.5%). Nine major patterns were determined by PFGE. In 6 of these patterns, thirty-six strains (90%) had identical PFGE profiles.

Detection of beta-lactam antibiotic resistant genes in Escherichia coli from porcine fecal samples using DNA chip

  • Park, Nam-Yong;Na, Sung-Ho;Cho, Ho-Seong
    • 한국동물위생학회지
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    • 제30권4호
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    • pp.505-510
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    • 2007
  • This study was conducted to detect ${\beta}$-lactam antibiotic-resistant genes in the 400 E coli isolates from porcine fecal samples in Korea by a DNA chip. The DNA chip contains the specific probe DNAs of the ${\beta}$-lactam antibiotic-resistant genes that had been labeled with a mixture of primer set designed to amplify specific genes (PSE, OXA, FOX, MEN, CMY, TEM, SHV, OXY and AmpC) using a multiplex polymerase chain reaction (PCR). Of 400 isolates 339 contained at least one ${\beta}$-lactamases gene. Resistance to ${\beta}$-lactamases was mediated mainly by AmpC (n = 339, 100%), and followed by TEM (n = 200, 59.0%), CMY (n = 101, 29.8%), PSE (n = 30, 8.9%) and both OXA and SHV genes (n = 20, 5.9%), while the FOX, MEN and OXY genes were not detected. The other sixty-one did not contain any ${\beta}$-lactamase genes even though they were resistant to antimicrobial drugs. In conclusion, the DNA chip system can be used as a rapid and reliable method for detecting of ${\beta}$-lactamases genes, which will help veterinarians select the antibiotics for monitoring and treating of animal diseases.

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

  • 김윤경;홍해숙
    • 대한간호학회지
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    • 제37권6호
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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.

A Retrieved Sparganum of Spirometra erinaceieuropaei from a Korean Man during Mechanical Thrombectomy

  • Hwang, Yang-Ha;Son, Wonsoo;Kim, Yong-Won;Kang, Dong-Hun;Chang, Hyun-Ha;Goo, Youn-Kyoung;Hong, Yeonchul;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • 제58권3호
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    • pp.309-313
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    • 2020
  • Human sparganosis is a zoonotic disease caused by infection and migration of the plerocercoid of Spirometra spp. Although sparganosis were reported from most parts of the body, the sparganum parasitizing inside cerebral artery is remarkably uncommon. We report a case of cerebral intravascular sparganosis in an elderly patient with acute ischemic stroke who was diagnosed by retrieving sparganum during mechanical thrombectomy. Finally, the parasites were identified as Spirometra erinaceieuropaei using multiplex PCR and cox1 gene sequencing.

Molecular fingerprinting of olive flounder pathogenic Streptococcus parauberis strains by random amplified polymorphic DNA analysis

  • Jung, Yong-Uk;Kang, Sang-Hyuck;Jin, Chang-Nam;Kang, Bong-Jo;Heo, Moon-Soo
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2005년도 생물공학의 동향(XVII)
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    • pp.899-903
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    • 2005
  • Two infectious species of Streptococcosis pathogens were detected by multiplex PCR assay. Detection rates of Streptococcus iniae and S. parauberis could reach 44.9% and 55.1% respectively for one year during 2004 to 2005 in Jeju island. These findings showed that S. parauberis strains were important pathogen with streptococcosis of olive flounder in Jeju island. These findings showed that S. parauberis strains were important pathogen with streptococcosis of olive flounder in Jeiu island. In the present study we have investigated the interspecific relationship of all Jeju area of S. parauberis by RAPD analysis. Represent strains divided to four groups by RAPD fingerprints. The important differences observed between the olive flounder isolates suggest that they could constitute a well-differentiated group or a separate clonal line within this bacterial species. Though, serological research of S. parauberis strains in Jeju island not exist yet. These strains doing the serological evolution.

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Mitochondrial nad 7 intron 4 region을 통한 분자생물학적 고려인삼품종 "천풍"검증 (Molecular identification of Korean ginseng cultivar "Chunpoong" using the mitochondrial nad 7 intron 4 region)

  • 왕홍도;김민경;권우생;양덕춘
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2010년도 정기총회 및 춘계학술발표회
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    • pp.15-15
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    • 2010
  • Koran ginseng(Pnax ginseng) is one of the most important medicinal plants in Orient. Among the nine cultivars of Korea ginseng, Chunpoong commands a much greater market value and has been planted widely. A rapid and reliable method for discriminating the Chunpoong cultivar was developed by exploiting a single nucleotide polymorphism (SNP) in the mitochondrial nad7 intron 4 region of nine Korea ginseng cultivars using universal primers. A SNP was detected between Chunpoong and other cultivars and modified allele-specific primers were designed from this SNP site to effective method for the geneic identification of the Chunpoong cultivar of ginseng.

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