• Title/Summary/Keyword: molecular typing

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Antibiotic Susceptibility Pattern and Molecular Typing By PCR-RAPD Analysis of Clinical and Environmental Isolates of Pseudomonas aeruginosa

  • Oluborode, O.B.;Smith, S.I.;Seriki, T.A.;Fowora, M.;Ajayi, A.;Coker, A.O.
    • Microbiology and Biotechnology Letters
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    • v.46 no.4
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    • pp.434-437
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    • 2018
  • Pseudomonas aeruginosa accounts for a significant proportion of nosocomial infections. This study examined the antimicrobial susceptibility pattern and clonal relatedness of P. aeruginosa isolates of clinical and environmental origin. These isolates displayed susceptibility to levofloxacin, ciprofloxacin, gentamicin, imipenem, and ceftazidime of 65.0%, 62.5%, 90.0%, 100%, and 85%, respectively. PCR-RAPD analysis of the P. aeruginosa isolates revealed marked variation. No correlation was observed between the antibiotic resistance profiles and the DNA typing patterns.

Rapid Typing of Clinical Strains of Mycobacterium tuberculosis by IS6110-based Outward PCR

  • Kim, Yeun;Lee, Uen-Ho;Park, Young-Kil;Bai, Gill-Han;Cho, Sang-Nae;Lee, Hye-Young
    • Biomedical Science Letters
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    • v.10 no.2
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    • pp.163-169
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    • 2004
  • Worldwide, tuberculosis remains one of the leading infectious diseases, accounting for nearly 3 million deaths and more than 8 million new cases annually. DNA typing of Mycobacterium tuberculosis is important for the control of tuberculosis, since it can be used to track transmission route of tuberculosis, source of internal laboratory contaminations, and to answer questions on the nature of tuberculosis infections such as reactivation or exogenous reinfection of disease. At present, IS6110-based RFLP is the choice of method for typing large numbers of clinical isolates of M. tuberculosis, since it has the highest resolution power. However, RFLP requires long time, high cost and qualified experts, so only reference level laboratories can use the RFLP technique. In order to have an optional molecular typing method suitable for the clinical settings, this study evaluated the use of one of PCR-based typing methods, IS6110-based outward PCR for typing clinical isolates of M. tuberculosis. In brief, the results from this study showed that IS6110-based RFLP is useful to discriminate diverse clinical isolates of M. tuberculosis as well as to identify clinical isolates that belong to the same family or cluster groups that have been previously classified by RFLP analysis. In addition, the banding profiles resulted from IS6110-based outward PCR seemed to represent genomic characteristics of M. tuberculosis, since strains belong to the K-family generated unique band that is not present in any other strains but present only in the genome of K-family strains. The IS6110-based outward PCR was also shown to be useful with DNAs isolated directly from liquid cultures indicating this method can be suitable for typing M. tuberculosis in clinical settings.

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Enhanced Discrimination of Listeria spp. Using RAPD Fingerprinting Complemented by Ribotyping-PCR (리스테리아균의 특성분석을 위한 Molecular Typing 방법의 상호보완)

  • 임형근;홍종해;박경진;최원상
    • Journal of Life Science
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    • v.13 no.5
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    • pp.699-704
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    • 2003
  • The results typed by random amplification of polymorphic DNA (RAPD) were compared with those obtained by Enterobacterial repititive intergenic consensus (ERIC) fingerprinting and ribotyping-PCR. The discriminatory power of RAPD typing was the best among the methods tested. RAPD typing with two different primers for 13 Listeria spp. reference strains produced 11 patterns each. In contrast, ERIC fingerprinting produced 9 patterns and ribotyping-PCR produced 7 patterns each. Composite of two separate RAPD (Lis 11 and primer 6) results or RAPD (Lis11)/ ribotyping-PCR differentiated all 13 Listeria spp. reference strains. Therefore, composite of 2 separate RAPD (Lis11 and primer 6) or composite of RAPD (Lis11)/ribotyping-PCR is expected the most promising approach for typing field isolated Listeria spp. strains.

Molecular Genetic Characteristics of Methicillin-Resistant Staphylococcus aureus Isolated from Patients and Environment of General Hospital Intensive Care Unit in a Chungnam Province, Korea (충남지역 종합병원 중환자실 환경과 환자로부터 분리한 메티실린 내성 황색포도알균(MRSA)의 분자유전학적 특성)

  • Kim, Hye-Suk;Park, Sung-Bae;Kim, Sang-Ha;Kim, Sunghyun;Hyun, Sung-Hee;Kim, Young-Kwon
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.2
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    • pp.110-117
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    • 2018
  • In the present study, mec complex typing and SCCmec typing were performed to analyze the molecular genetic characteristics of 20 methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from clinical specimens and 4 strains isolated from the ICU environments of secondary medical institutions in a Chungnam province, Korea, from June to July of 2017. Among a total of 20 MRSA strains isolated from clinical specimens, 8 cases (40%) were SCCmec type II, one case (5%) was SCCmec type IVa, and 11 cases (55%) were not-typeable in SCCmec type analysis. Among 4 MRSA isolates from the ICU environment, one strain did not have the mecA gene and 3 strains were typed as SCCmec types II, III, and IVa, respectively. Data from the present study showed that the origin of MRSA isolated from the clinical specimens was different from those from the ICU environment in most cases but the origin was concordant in one case. In this case, MRSA might be transmitted by healthcare workers to the ICU environment. Further study with a large number of cases and other hospital infection-related microorganisms will be needed. This continuous follow-up study might provide useful information on infection control in medical institutions.

Molecular Epidemiology of Cryptococcus neoformans/Cryptococcus gattii Complex Isolates from Pigeon Droppings in Korea

  • Chang, Kyungsoo
    • Biomedical Science Letters
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    • v.19 no.3
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    • pp.213-223
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    • 2013
  • The objectives of this study are to develop a molecular diagnosis to differentiate serotypes and mating-types of C. neoformans/C. gattii complex isolates from pigeon droppings in Korea and to elucidate molecular epidemiology of the isolates. Phenotypes and genotypes of C. neoformans/C. gattii complex isolates were identified by biochemical properties and PCR using specific CNLAC1 gene, respectively. To classify serotypes and mating-types of C. neoformans/C. gattii complex isolates, the five reference strains and thirty-three isolates in Korea were investigated by restriction fragment length polymorphism (RFLP) analysis using CNLAC1 gene for varieties, by random amplified polymorphic DNA (RAPD) for serotyping, and by PCR using specific primer sets for mating typing. All isolates in Korea were belonged to C. neoformans var. grubii (serotype A) by RFLP and RAPD patterns which showed high sensitivity and specificity. Therefore, RFLP and RFLP were available to differentiate varieties and serotypes of C. neoformans. Amplification patterns of the five reference strains by specific PCR for mating typing were differentiable, and all isolates were classified into $MAT{\alpha}$. All C. neoformans environmental isolates in Korea were Cr. neoformans serotype A and $MAT{\alpha}$ which is a more virulent pathogen. This study suggests that RFLP and RAPD are rapid and correct molecular diagnosis tools for epidemiology of C. neoformans/C. gattii complex isolates.

Random amplification of polymorphic DNA typing of Listeria monocytogenes isolates from animal products (축산물유래 Listeria monocytogenes의 RAPD typing)

  • Lee, Chul-Hyun;Son, Won-Geun
    • Korean Journal of Veterinary Research
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    • v.49 no.4
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    • pp.319-328
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    • 2009
  • This study investigated the epidemiology of Listeria (L.) monocytogenes, a food-borne pathogen. The epidemiology of food-borne pathogens is of great importance for clarifying bacterial origin and preventing bacterial contamination and infection. This work examined 68 L. monocytogenes strains, including 11 reference strains and 57 isolates from imported US beef, domestic meats (beef, pork, chicken meat), raw milk, and milk plants. The random amplified polymorphic DNA (RAPD) techniques were optimized to develop a standard molecular epidemiological analysis of L. monocytogenes. There was great genetic variability among the isolates, which produced 24 and 34 RAPD patterns with primer HLWL85 and HLWL74, respectively. The discriminatory power of the RAPD methods with HLWL85 and HLWL74 primer were very high (DI = 0.957; S ${\geq}$ 80%, S ${\geq}$ 95%). Some RAPD types were specific to origin. A few RAPD types were specific for L. monocytogenes strains belonging to a particular serotype. Using the HLWL85 primer, the strains isolated from milk plants could be distinguished from the other strains. And using the HLWL74 primer, the strains isolated from imported beef (US) could be distinguished completely from the other strains.

Optimal Combination of VNTR Typing for Discrimination of Isolated Mycobacterium tuberculosis in Korea

  • Lee, Jihye;Kang, Heeyoon;Kim, Sarang;Yoo, Heekyung;Kim, Hee Jin;Park, Young Kil
    • Tuberculosis and Respiratory Diseases
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    • v.76 no.2
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    • pp.59-65
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    • 2014
  • Background: Variable-number tandem repeat (VNTR) typing is a promising method to discriminate the Mycobacterium tuberculosis isolates in molecular epidemiology. The purpose of this study is to determine the optimal VNTR combinations for discriminating isolated M. tuberculosis strains in Korea. Methods: A total of 317 clinical isolates collected throughout Korea were genotyped by using the IS6110 restriction fragment length polymorphism (RFLP), and then analysed for the number of VNTR copies from 32 VNTR loci. Results: The results of discriminatory power according to diverse combinations were as follows: 25 clusters in 83 strains were yielded from the internationally standardized 15 VNTR loci (Hunter-Gaston discriminatory index [HGDI], 0.9958), 25 clusters in 65 strains by using IS6110 RFLP (HGDI, 0.9977), 14 clusters in 32 strains in 12 hyper-variable VNTR loci (HGDI, 0.9995), 6 clusters in 13 strains in 32 VNTR loci (HDGI, 0.9998), and 7 clusters in 14 strains of both the 12 hyper-variable VNTR and IS6110 RFLP (HDGI, 0.9999). Conclusion: The combination of 12 hyper-variable VNTR typing can be an effective tool for genotyping Korean M. tuberculosis isolates where the Beijing strains are predominant.

Molecular Typing of Leuconostoc citreum Strains Isolated from Korean Fermented Foods Using a Random Amplified Polymorphic DNA Marker

  • Kaur, Jasmine;Lee, Sulhee;Sharma, Anshul;Park, Young-Seo
    • Food Engineering Progress
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    • v.21 no.2
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    • pp.174-179
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    • 2017
  • For preliminary molecular typing, PCR-based fingerprinting using random amplified polymorphic DNA (RAPD) is the method of choice. In this study, 14 bacterial strains were isolated from different Korean food sources, identified using 16S rRNA gene sequencing, and characterized through RAPD-PCR. Two PCR primers (239 and KAY3) generated a total of 130 RAPD bands, 14 distinct PCR profiles, 10 polymorphic bands, one monomorphic band, and four unique bands. Dendrogram-based analysis with primer 239 showed that all 14 strains could be divided into seven clades out of which clade VII had the maximum of seven. In contrast, dendrogram analysis with the primer KAY3 divided the 14 L. citreum strains into four clades out of which clade IV consisted of a maximum of 10 strains out of 14. This research identified and characterized bacterial populations associated with different Korean foods. The proposed RAPD-PCR method, based on sequence amplification, could easily identify and discriminate the lactic acid bacteria species at the strain-specific level and could be used as a highly reliable genomic fingerprinting tool.

Prevalence and Characteristics of Antimicrobial-Resistant Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus from Retail Meat in Korea

  • Kim, Yong Hoon;Kim, Han Sol;Kim, Seokhwan;Kim, Migyeong;Kwak, Hyo Sun
    • Food Science of Animal Resources
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    • v.40 no.5
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    • pp.758-771
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    • 2020
  • This study was to investigate the prevalence and characteristics of antimicrobial-resistant Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) from 4,264 retail meat samples including beef, pork, and chicken in Korea between 2013 and 2018. A broth microdilution antimicrobial susceptibility testing was performed for S. aureus. Molecular typing by multilocus sequence typing (MLST), spa typing, and pulsed-field gel electrophoresis (PFGE), was performed on mecA-positive S. aureus strain. S. aureus was isolated at a rate of 18.2% (777/4,264), of which MRSA comprised 0.7% (29 strains). MLST analysis showed that 11 out of the 29 MRSA isolates were predominantly sequence type (ST) 398 (37.9%). In addition, ST72, ST692, ST188, ST9, and ST630 were identified in the MRSA isolates. The spa typing results were classified into 11 types and showed a high correlation with MLST. The antimicrobial resistance assays revealed that MRSA showed 100% resistance to cefoxitin and penicillin. In addition, resistance to tetracycline (62.1%), clindamycin (55.2%), and erythromycin (55.2%) was relatively high; 27 of the 29 MRSA isolates exhibited multidrug resistance. PFGE analysis of the 18 strains excluding the 11 ST398 strains exhibited a maximum of 100% homology and a minimum of 64.0% homology. Among these, three pairs of isolates showed 100% homology in PFGE; these results were consistent with the MLST and spa typing results. Identification of MRSA at the final consumption stage has potential risks, suggesting that continuous monitoring of retail meat products is required.