• 제목/요약/키워드: RAPD fingerprints

검색결과 18건 처리시간 0.026초

Molecular Markers for Identification of Stellantchasmus falcatus and a Phylogenic Study using the HAT-RAPD Method

  • Wongsawad, Chalobol;Wongsawad, Pheravut
    • Parasites, Hosts and Diseases
    • /
    • 제48권4호
    • /
    • pp.303-307
    • /
    • 2010
  • Stellantchasmus falcatus is a minute intestinal fluke in the family Heterophyidae. Metacercariae, the infective stage, were reported in a marine fish, mullet Liza subviridis, and a fresh water fish, Dermopgenus pusillus, in Thailand. Adults were found in chicks, rats, cats, and humans. Morphological studies were done for comparing Stellantchasmus sp. worms found in 2 different fish hosts; their shapes and organ arrangements were very similar except for the prepharynx length. Therefore, the present study aimed to compare their DNA fingerprints using the HAT-RAPD method for both types of Stellantchasmus and several other related species. Ten arbitrarily selected primers (OPA-04, OPA-09, OPN-02, OPN-03, OPN-09, OPN-12, OPP-11, OPR-15, OPX-13, and OPAD-01) were used. It was found that OPA-09, OPN-03, and OPAD-01 were able to generate S. falcatus specific fragments in mullets which consisted of 200, 760, and 280 bp, respectively. In addition, the results of morphologic, DNA fingerprinting, and phylogenetic analyses strongly suggest that the fresh water and marine specimens of Stellantchamus may be different species.

넙치 (Paralichthys olivaceus) 병어에서 분리된 연쇄상구균의 다양성 (Diversity of the Streptococcal Strains Isolated from Diseased Olive Flounder (Paralichthys olivaceus))

  • 김종훈;김은희
    • 한국수산과학회지
    • /
    • 제36권6호
    • /
    • pp.654-660
    • /
    • 2003
  • To evaluate the biological diversity of fish pathogenic streptococci, 35 strains isolated from diseased olive flounder (Paralichtys olivaceus), were analyzed using a random amplified polymorphic DNA (RAPD) technique with the oligonucleotide commercial primer 6 (Amersham Biosciences). Api 20 Strep test, drug resistance and artificial infection were carried out for further characterization of the isolates. RAPD fingerprints showed similar pattern in 25 strains (about $71.4\%$ of 35 isolates) and these strains were designed as RA group 1. Similarities greater than $44\%$ were obtained when the Dice coefficient was applied among the isolates of RA 1. On the other hand, the reference Streptococcus iniae showed a similar RAPD profile to the isolates with similarity levels of $40-93.3\%.$ Rh I was suggested to be the dominant group isolated from olive flounder suffering from streptococcosis. However, the isolates of Rh 1 group were not classified into the same species by the Api 20 Strep identification system. There was no peculiarity in drug resistance patterns of Rh I group isolates against 7 antibacterial agents. However, only 3 of 25 isolates $(0.12\%)$ showed oxytetracycline (OTC) resistance and OTC might be a useful chemotherapeutic agent in controlling the streptococcosis by strains of RA I group in olive flounder. Fish injected intraperitoneally with $10^5$ CFU of an isolate of Rh I and RA III group showed $60\%\;and\;50\%$ accumulative mortality for 20 days, respectively ($20\%$ in control or Rh II). However luther comparative studies about differences in virulence between isolates are needed.

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
    • /
    • 한국생물공학회 2005년도 생물공학의 동향(XVII)
    • /
    • pp.899-903
    • /
    • 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.

  • PDF

S-haplotypes and Genetic Diversity in 'Danji' Radish (Raphanus sativus L. var. hortensis)

  • Ahn, Yulkyun;Kim, Hyukjun;Han, Dongyeop;Park, Younghoon
    • 원예과학기술지
    • /
    • 제32권2호
    • /
    • pp.210-216
    • /
    • 2014
  • The distribution of S-haplotypes and genetic relationships were evaluated for 47 accessions of 'Danji' radish (Raphanus sativus L. var. hortensis Baker f. gigantissimus Makino) originating from Jeju Island in South Korea. A total of 22 S-haplotype-specific SCAR markers for the S locus glycoprotein (SLG) and S receptor kinase (SRK) loci were tested, and six primer sets amplified locus-specific PCR fragments from at least one 'Danji' radish accession. S5 and S21 alleles atthe SLG locus were the most frequently distributed, and detected from 87.5% and 64.6% of the accessions, respectively. The frequency of the class-II haplotype at the SLG locus was 75%, more frequent than the class-I haplotype. The S23 allele at the SRK locus was detected from 7 accessions. Grouping of the accessions based on S-allele composition revealed three major groups, while 8 accessions showed a unique allelic composition. The genetic diversity of 47 'Danji' radishes and 1 'Gwandong' radish were also evaluated with 38 RAPD primers. A total of 312 bands were scored, and showed that 138 bands (44.2%) were monomorphic among the accessions, whereas 174 (55.8%) bands were polymorphic. Polymorphism rates ranged from 0.2 to 1.0, indicating significant variations in detecting polymorphism across RAPD primers. The genetic similarity coefficients among all pairs of the 48accessions varied from 0.62 to 0.93, and 42% of the comparisons exhibited values higher than 0.85. All the cultivars could be distinguished based on the DNA fingerprints revealed by RAPD. The comparisons between the dendrograms based on S-haplotypes and RAPDs indicate an unrelated and sporadic distribution for several accessions; however, there was a tendency for accessions with the same S-allelic composition to group into the same cluster.

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
    • 한국미생물·생명공학회지
    • /
    • 제49권2호
    • /
    • pp.239-248
    • /
    • 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.

Susceptibility Testing of Helicobacter pylori to Metronidazole and DNA Fingerprints of Resistant Strains in Singapore

  • HUA, JIESONG;KHAY-GUAN YEOH;PENGYUAN ZHENG;HAN CHONG NG;BOW HO
    • Journal of Microbiology and Biotechnology
    • /
    • 제9권3호
    • /
    • pp.328-333
    • /
    • 1999
  • Susceptibility of 61 strains of Helicobacter pylori to metronidazole was examined by both the disk diffusion method using a cut-off of 15㎜ for resistance and the E test with a cut-off of 8㎎/l. The MIC/sub 50/ and MIC/sub 90/ by the E test were 2 ㎎/l and 256㎎/l, respectively. Metronidazole resistance was found in 22 (36%) out of the 61 H. pylori strains by the E test and in three additional strains by the disk diffusion method. Amongst the latter three isolates, the MICs by the E test were 4 ㎎/l, 6㎎/l, and 6㎎/l, respectively. These figures are one log₂ or half log₂ dilution lower than the cut-off of 8㎎/l recommended as resistance for the E test. All 22 metronidazole resistant H. pylori isolates by the E test that were subjected to random amplified polymorphic DNA (RAPD) fingerprinting showed different DNA fingerprints. Interestingly, >90% of resistant isolates possess two common DNA bands of 0.4 and 0.9 kb. This study demonstrates that the results of the disk diffusion method for testing H. pylori susceptibility to metronidazole correlates well with that of the E test. The criteria for interpretation need to be internationally standardized so that the results from different centers can be compared.

  • PDF

Specific and Sensitive Detection of Venturia nashicola, the Scab Fungus of Asian Pears, by Nested PCR

  • Koh, Hyun Seok;Sohn, San Ho;Lee, Young Sun;Koh, Young Jin;Song, Jang Hoon;Jung, Jae Sung
    • The Plant Pathology Journal
    • /
    • 제29권4호
    • /
    • pp.357-363
    • /
    • 2013
  • The fungus Venturia nashicola is the causal agent of scab on Asian pears. For the rapid and reliable identification as well as sensitive detection of V. nashicola, a PCR-based technique was developed. DNA fingerprints of three closely related species, V. nashicola, V. pirina, and V. inaequalis, were obtained by random amplified polymorphic DNA (RAPD) analysis. Two RAPD markers specific to V. nashicola were identified by PCR, after which two pairs of sequence characterized amplified region (SCAR) primers were designed from the nucleotide sequences of the markers. The SCAR primer pairs, designated as D12F/D12R and E11F/E11R, amplified 535-bp and 525-bp DNA fragments, respectively, only from genomic DNA of V. nashicola. The specificity of the primer sets was tested on strains representing three species of Venturia and 20 fungal plant pathogens. The nested PCR primer pair specific to V. nashicola was developed based on the sequence of the species-specific 525-bp DNA fragment amplified by primer set E11F/E11R. The internal primer pair Na11F/Na11R amplified a 235-bp fragment from V. nashicola, but not from any other fungal species tested. The nested PCR assay was sensitive enough to detect the specific fragment in 50 fg of V. nashicola DNA.