• Title/Summary/Keyword: RAPD PCR

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Genotyping and Molecular Characterization of Carbapenem-resistant Acinetobacter baumannii Strains Isolated from Intensive Care Unit Patients

  • Abozahra, Rania;Abdelhamid, Sarah M.;Elsheredy, Amel G.;Abdulwahab, Kawther E.;Baraka, Kholoud
    • Microbiology and Biotechnology Letters
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    • v.49 no.2
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    • pp.239-248
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    • 2021
  • The emergence of multidrug-resistant Acinetobacter baumannii has partly increased treatment failure and patient mortality. Class D β-lactamases is an important mechanism of resistance to beta-lactam antibiotics in this species. This study aimed to investigate the relationship between the presence oxacillinase gene and genetic fingerprints of A. baumannii isolates from the intensive care unit of an Egyptian tertiary care hospital. One hundred and twenty A. baumannii clinical isolates were collected. Multiplex PCR was performed to detect genes encoding oxacillinases (OXA-23, OXA-24, OXA-51, OXA-58 and OXA-143). Molecular typing of all collected isolates was performed using random amplified polymorphic DNA (RAPD)-PCR assay. Out of 120 examined isolates, 92, 88 and 84% were resistant to ertapenem, imipenem and meropenem, respectively. The species-specific, commonly present OXA-51 gene was found in all isolates while OXA-23 showed a high prevalence of 88% of isolates. OXA-24 and OXA-143 genes were detected in 3% and 1% of isolates, respectively. No OXA-58 gene was detected. Five clusters consisting of 19 genotypes were detected using RAPD-PCR. Genotype A was the most prevalent, it was observed in 62% of the isolates followed by genotype B (12%). These results revealed that genotypes A and B are common in the hospital. Results also demonstrate that RAPD-PCR is a rapid and reliable method for studying the clonal similarity among A. baumannii isolated from different clinical specimens.

Analysis of Genetic Relationship by RAPD Technique for Codonopsis lanceolata Trauty Collected from the Baekdoo Mountain and Korea (백두산지역과 국내 더덕 수집종의 RAPD에 의한 유연관계 분석)

  • Doo, Hong-Soo;Ryu, Jeom-Ho;Lee, Kang-Soo;Li, Hu Lin;Liu, Xian Hu
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.3
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    • pp.194-199
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    • 2002
  • Extracted genomic DNA from 16 accessions of Codonopsis lanceolata collected from South Korea and the Baekdoo Mt. areas of China were analyzed for their genetic relationships by RAPD. Twenty 10-mer-oligonucleotide primers having reproductive polymorphism were selected for the RAPD analysis. The size of amplified DNA was almost between 125 bp and 2.0 kbp. Sixteen collected Codonopsis lanceolata were analyzed with 20 primers which generated 73(49.3%) polymorphic bands among 148 PCR products. The mean number of polymorphic bands were 7.4 and varied $1{\sim}9$ per primer. It was, thus, demonstrated that RAPD was useful for detecting polymorphism in Codonopsis lanceolata. The range of 1-F value(genetic similarity) was from 0.682 to 0.959. These results indicate variable genetic similarities. By UPGMA (Unweighted Pair Group Method using an Arithmetic average) cluster analysis based on 1-F value, genetic distance among the 16 collected Codonopsis lanceolata was $0.133{\sim}0.400$. It was certainly classified into two groups between collected accessions from Korea and China, and the genetic distance was about 0.281. Both accessions collected from Korea and China showed miner differences, while the genetic relationships of Tonghua Xian and Liuhe Xian from China was farthest with other accessions collected.

Development and Application of PCR-based Markers for the Discrimination of Bang-Poong and Related Species (방풍류의 감별을 위한 분자마커의 탐색과 활용)

  • Hong, Seong-Mi;Lee, Mi-Young;Koh, Jae-Chul;Ko, Byoung-Soeb
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.1-6
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    • 2004
  • Bang-Poong and related species are an important herbal medicine. However, it is difficult to determine the commercial dry material through anatomical and chemotaxonomical characteristics. Here, we used a PCR-based technique for an accurate discrimination of Bang-Poong and related species. With the RAPD primers, 215 RAPDSs(random amplified polymorphic DNAs) were obtained, and 98% of them showed polymorphic patterns. RAPDs from the four primers were appropriate for the discrimination of S. divaricata $(T_{URCZ{\cdot}})\;S_{CHISKIN}$, those from the six primers for P. japonicum $T_{HUNBERG}$, those from the four primers for P. terebinthaceum $F_{ISHER}$, and those from the six primers for G. littoralis Fr. $S_{CHMIDT}$. The specific bands from the primer 425 were obtained and used to develop SCAR (sequence characterized amplified region) markers, based on the sequence information of the RAPD markers. The SCAR primers generated a 215 bp fragment specific to Peucedanum terebinthaceum $F_{ISHER}$, and a 177 bp and a 300 bp fragment specific to G. littoralis Fr. $S_{CHMIDT}$. As a result, the three SCAR markers were able to discriminate from two Bang-Poong related species.

Genetic Relationships among the Parental Bombyx mori Strains of the Current F$_1$ Hybrid Silkworm based on RAPD (RAPD를 이용한 장려누에품종의 원종간 유전적 유연관계)

  • 황재삼;이진성;강현아;이상몽;손해룡
    • Korean journal of applied entomology
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    • v.36 no.3
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    • pp.206-214
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    • 1997
  • The genetic relationships among the twenty parental silkworm, Bombyx mori strains authorized in Korea were evaluated using RAPDs-PCR(Random Amplified Polymorphic DNAs-Polymerase Chain Reaction). Twenty-six different 10-mer oligonucleotide primers were used to screen genetic characteristics of parental twenty silkworm strains by RAPD-PCR analysis. 24 primers showed different banding patterns among the strains. Based on these RAPD patterns, the genetic relationships among the silkworm strains were analyzed by UPGMA(Unweighed Pair-Group Method with Arithmetic average) method. The phylogenetic relationships in the twenty silkworm strains were classified into two major sub-groups at the genetic similarity coefficient of 0.60. The first sub-group included Jaml13, Jaml 19, Jaml20, Jam123, Jam1 25 and Jam 127. Jamll4, Jam1 2 I, Jam 122, Jam 124, Jam1 26, Jam 128, Jam129, Jam 130, Jam 131, Jam1 32, Jam133, Jam134, Jam301 and Jam302 were included in the 2nd group. The genetic distance values among Jam1 14, Jam120 and Jam127 were lower than those among the other strains, while Jam129 is very closely related to Jam131 as showing coefficient value of 1 .O.

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Molecular Phylogeny of Korean-type Coliphages and American-type Coliphages Determined by a RAPD Analysis (RAPD 분석법에 의한 한국형 대장균파아지와 미국형 대장균파아지의 분자적 계통분류)

  • 권오식
    • Biomedical Science Letters
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    • v.6 no.2
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    • pp.131-139
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    • 2000
  • RAPD-PCR was applied to identify the phylogenetic relationship between isolated Korean-type coliphages ($\phi$C1, $\phi$C2, $\phi$C3 and $\phi$C4) and well-known American coliphages ($\phi$T2, $\phi$T4, $\phi$T5, $\phi$T7 and ${\phi}{\lambda}$). Subsequently, a computer analysis was carried out with the results of RAPD-PCR. As a result, 9 individuals were divided into five groups. The Korean-type coliphages formed a single cluster which showed very high genetic similarity but the American-type coliphages revealed very low genetic similarity among them. In particular, the $\phi$T2와 $\phi$T4 (T$_{even}$ phages) made one sub-cluster among American coliphages, and they were very distant from $\phi$T5, $\phi$T7 and ${\phi}{\lambda}$. However, ${\phi}{\lambda}$ made a cluster with the Korean-type coliphages that we isolated. The genome size of Korean-type coliphages was ranged from 25,000 bp to 35,000 bp. Among them, the genome of $\phi$C2 was the smallest and that of $\phi$C1 was the biggest, while others were in the middle of the size.

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Analysis genetic diversity of Plasmodiophora brassicae using RFLP and RAPD(oral)

  • Heo, Seung-Hwan;Jang, Chang-Soon;Lee, Hyoun-Kyoung;Lee, Woo-Chung;Jang, Se-Jeong;Kim, Hong-Gi
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.112.1-112
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    • 2003
  • Genetic diversity of Plasmodiophora brassicae from major chinese cabbage cultivating areas in Korea was analyzed by using PCR-RFLP and RAPD. Single spores of P brassicae isolated from galls of club root made induce lesion on chinese cabbage successfully. The PCR-RFLP and RAPD by primers PbITS, URP 3, 6 and OPA 7 revealed that single spore isolates showed various DNA polymorphisms among them unrelated geographic origins. These results indicate that P. brassicae population in Korea showed genetic difference among them. This study could be facilitate to identify genetic characteristics ofP. brassicae based on DNA polymorphisms between single spore isolates and to get basic information which can be used to advanced resistance breeding against club root of chinese cabbage.

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Genetic Relationships of Lactuca spp. Revealed by RAPD, Inter-SSR, AFLP, and PCR-RFLP Analyses

  • Yang, Tae-Jin;Jang, Suk-Woo;Kim, Won-Bae
    • Journal of Crop Science and Biotechnology
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    • v.10 no.1
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    • pp.27-32
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    • 2007
  • RAPD, Inter-SSR, and AFLP markers were used to assess the genetic diversity of lettuce cultivars and the phylogenetic relationships in Lactuca spp. A total of 216 polymorphic bands from seven RAPD primers, four Inter-SSR primers, and five AFLP primer combinations were used to elucidate the genetic similarity among lettuce cultivars. Forty-four lettuce accessions were subdivided into discrete branches according to plant type: crisphead, butterhead, and stem type, with some exceptions. The leafy- and cos-type accessions were intermingled in other groups with no discrete branch indicating that these are more diverse than others. Three accessions, including the Korean cultivar 'Cheongchima', the Korean local landrace 'Jinjam', and the German cultivar 'Lolla Rossa' were classified as the most diverse accessions. Twenty bands were unique in specific cultivars. Among these, three were specific in a plant type; one in Korean leafy type, one in crisphead type, and one in cos type lettuce. In the phylogenetic analysis among Lactuca species, L. saligna, L. serriola, and L. georgica clustered in a sister branch of the L. sativa complex. Two L. virosa accessions show the highest intra-specific relationships. L. perennis outlied from all the other Lactuca species at a genetic similarity of 0.53 and clustered with two Cichorium species, C. intybus and C. endivia, with genetic similarity of 0.67. The phylogenetic tree was supported by data from polymorphism of chloroplast genome which was revealed by PCR-RFLP.

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A Systematic Relationship of the Korean Symplocaceae Based on RAPD analysis (RAPD에 의한 한국산 노린재나무과 식물의 유연관계 분석)

  • Park, Sang-Hong;Lee, Joongku;Kim, Joo-Hwan
    • Korean Journal of Plant Taxonomy
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    • v.37 no.3
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    • pp.225-237
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    • 2007
  • In order to estimate the genetic relationships among four taxa of Korean Symplocaceae and their regional populations, RAPDs analysis was performed. The length of the amplified DNA fragments ranged from 150 to 1,900 bp. Ninety-two RAPD bands were scored for 11 primers and the genetic distance was calculated with by Nei-Li's genetic dissimilarity. Deciduous and evergreen groups were clearly separated in the UPGMA analysis. Symplocos coreana was clustered as a distinct group, and S. sawafutagi and S. tanakana were also clustered at specific level, respectively. The RAPD data was useful to cognize the genetic variation and to discuss the relationships among Korean Symplocaceae

Random Amplified Polymorphic DNA Analysis for Origin Identification of Olive Flounder (Paralichthys olivaceus) and Redlip Croaker (Pseudosciaena polyactis) (넙치와 조기의 원산지 판별을 위한 random amplified polymorphic DNA 패턴 연구)

  • Kang Duk-Jin;Lee Suk-Keun;Jin Deuk-Hee;Choi Suk-Jung
    • Journal of Life Science
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    • v.16 no.1
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    • pp.88-94
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    • 2006
  • The random amplified polymorphic DNA (RAPD) technique was investigated as a potential tool for the origin identification of olive flounder (Paralichthys olivaceus) and redlip croaker (Pseudosciaena polyactis). Olive flounder specimens were collected from North Korea and several locations of South Korea (Jumunjin, Tongyoung and Geoje). Fishes obtained from Tongyeong and Geoje were cultured products. Redlip croaker specimens were collected from South Korea and China. Consistent and distinct diagnostic bands were easily identified in the RAPD patterns of the olive flounder specimens. Although consistent diagnostic bands rarely appeared in the RAPD pattern of redlip croaker specimens because of their genetic heterogeneity, we were able to find potential diagnostic bands in the average RAPD pattern of each origin.

Identification of Phellinus linteus by Comparison of Colony Shapes and Using PCR techniques (목질진흙버섯(Phellinus linteus)의 균총형태 비교 및 PCR 기법을 이용한 동정)

  • Kong, Won-Sik;Kim, Dong-Hyun;You, Chang-Hyun;Kim, Young-Ho;Kim, Kyung-Soo;Kim, Kwang-Ho
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.466-477
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    • 1998
  • Twenty-two Phellinus strains were characterized using colony morphologies and polymerase chain reaction (PCR) to divide into Phellinus linteus. There were some differences in mycelial growth and colony shapes among the strains when they were grown on various media such as PDA, MCM, MEA and YM. Phellinus linteus was slowly growing, formed golden-yellow colony, and produced blue pigment on PDA media. When the regions of internal transcribed spacer (ITS) were amplified from ribosomal RNA (rRNA) coding genes of P. igniarius and P. linteus strains by means of PCR, two types of band (700 bp and 800 bp) were appeared, respectively. For the amplified intergenic region I (IGRI), P. igniarius strains showed a different band among 500, 600, 700 and 800 bp according to the strains, whereas P. linteus strains did one specific band of 700 bp. By polymorphism analysis after digesting the amplified products with 6 different restriction enzymes, a band specific to P. linteus was generated when the products for ITS region were digested with HaeIII, suggesting that the enzyme digestion could provide effective method to distinguish between P. igniarius and P. linteus. And also, the analysis of genetic relationship showed that the genetic similarities were 89% and 95% in P. igniarius and P. linteus strains, respectively. Random amplification polymorphic DNA (RAPD) analysis using multiple primer sets and arbitrarily primed PCR (AP-PCR) with ITS3 primer could also result in a reproducible way to identify P. linteus strains.

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