• 제목/요약/키워드: Multiplex-PCR

검색결과 492건 처리시간 0.029초

Detection of $\beta$-lactam Antibiotic-resistant Genes in Eschericia coli using DNA Chip from Porcine Fecal Samples

  • Na, Sung-ho;Cho, Ho-seong;Kim, Yong-hwan;A.W.E. Effendy;Park, Nam-yong
    • 한국수의병리학회:학술대회논문집
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    • 한국수의병리학회 2003년도 추계학술대회초록집
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    • pp.33-33
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    • 2003
  • There prevalence of $\beta$-lactamases bacteria in animals has been increased since 1990s [1]. The resistance in E coli which is mediated by $\beta$-lactamases hydrolyze the $\beta$-lactam ring eventually inactivate the antibiotics [2]. Generally, $\beta$-lactamases can be classified into four main groups and eight subgroups according to their functional and structural characteristics [3]. The detection of $\beta$-lactam antibiotic-resistant bacteria by DNA chip has been described [4]. The chip has a specific probe DNAs that contained the $\beta$-lactam antibiotic-resistant genes which was labeled by multiplex PCR reaction with a mixture of primer sets that were designed to amplify specific gene. Here we report the susceptibility of enteropathogenic E. coli isolated from pigs in Korea using the DNA chip in detecting $\beta$-lactam antibiotic-resistant genes. (omitted)

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말에서 분리한 Escherichia coli의 특성 및 항생제 감수성 (Characteristics and antimicrobial susceptibility of Escherichia coli isolated from horse)

  • 윤성욱;권도연;최성균;이희수;조길재
    • 대한수의학회지
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    • 제50권3호
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    • pp.231-237
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    • 2010
  • This study was conducted to investigate characteristics and antimicrobial susceptibility of Escherichia (E.) coli isolates isolated from vaginal mucosa and clitorial fossa of 105 Thoroughbred mares suspicious of the genital disease in Korea during the period from March 2006 to July 2007. Ninety six E. coli isolates were identified as standard biochemical properties and using BIOLOG system. Fifty three isolates (55.2%) could be classified into a total of 21 O serotypes and forty three isolates (44.8%) were non-typeable with 51 O antisera used in this study. The verotoxin 1 (VT 1) and verotoxin 2 genes were analyzed by multiplex PCR. Among them, one isolate was detected VT 1 gene (130 bp). Most of isolates showed a high susceptibility in ciprofloxacin (100%), enrofloxacin (100%), norfloxacin (100%), cefoxitin (96.9%), gentamicin (96.9%), sulphamethoxazole (96.9%), nitrofurantoin (94.8%), amikacin (93.8%), nalidixic acid (92.7%) and tetracycline (90.6%). These results may provide the basic information to establish strategies for the treatment and prevention of reproductive disease in Thoroughbred mares in Korea.

Characteristics and Pathogenicity for Japnaes Eel Anguilla japonica of Vibrio vulnificus Isolated from Oyster, Sediment and Seawater in the Korea Coast

  • Kim, Myoung Sug;Jung, Sung Hee;Hong, Suhee;Jeong, Hyun Do
    • Fisheries and Aquatic Sciences
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    • 제18권4호
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    • pp.387-393
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    • 2015
  • Biotyping of Vibrio vulnificus strains isolated from marine environments along the south coast of Korea showed that the majority of the isolates (94.7%) belonged to biotype 1 and the remaining isolates (5.3%) belonged to biotype 2. Analysis of 16S rRNA V. vulnificus strains isolated from marine environments using a multiplex polymerase chain reaction (PCR) revealed that 78.7% were type A and 21.3% were type B. Random amplified polymorphic DNA (RAPD) was used to analyze the genomic differences in V. vulnificus among the biotype 2 strains isolated from marine environments (newly isolated strains group) and reference strains obtained from infected eels (reference strains group). The two groups had distinctly different profiles of the amplicons produced from RAPD. Additionally, biochemical comparison of these strains revealed that all four strains isolated from marine environments differed from the strains isolated from eels in their ability to promote D-mannitol fermentation. Two (NH 1 and NH 2) out of four isolates of biotype 2 from marine environments showed pathogenicity in eels Anguilla japonica in a challenge test. These isolates did not agglutinate with antisera against V. vulnificus NCIMB 2137 (serovar E), ATCC 27562 (non-serovar E), and ATCC 33816 (atypical serovar E).

Tracing the origin of fish without hatchery information: genetic management of stock enhancement for mangrove red snapper (Lutjanus argentimaculatus) in Taiwan

  • Hsu, Te-Hua;Huang, Chang-Wen;Lin, Cheng-Hui;Lee, Hung-Tai;Pan, Chieh-Yu
    • Fisheries and Aquatic Sciences
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    • 제23권5호
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    • pp.13.1-13.7
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    • 2020
  • Stock enhancement is considered to be a valuable approach for restoring fishery resources. Because no specific official institution in Taiwan is responsible for the production of fry, the released fry are purchased directly from the private sector. However, fishermen from the private industry have not established a genetic background, so the genetic composition for each batch of released fry is unclear. Mangrove red snapper (Lutjanus argentimaculatus), a prominent species released in Taiwan, was collected after its official release. One hundred and two field samples were compared with four batches of hatchery fry (n = 685) by using a microsatellite-based multiplex PCR assay. Four of the field samples (3.9%; 4/102) were revealed to be from a fish farm and most likely from a single batch. This study revealed that wild mangrove red snappers are genetically different from those originating from farms, and their origins can be traced through molecular markers, even without information on breeding stocks.

약용작물의 기원 판별에 관한 분자생물학적 기술 개발 현황 (Development of molecular biological techniques for the differentiation of medicinal plant species)

  • 한은희;김윤희;이신우
    • Journal of Plant Biotechnology
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    • 제42권1호
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    • pp.6-12
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    • 2015
  • Medicinal plants resources are becoming important assets since their usages have been expanded to the development of functional foods for human health, more attractive cosmetics, and pharmaceutical industries. However, their phylogenetic origins and names are different from each country and quite often they are mixed each other resulting in the confusion for consumers. In particular, when they are very similar based on their morphological characteristics and distributed as dried roots, it is extremely difficult to differentiate their origins even by specialists. Recently, "DNA barcodes" have been extensively applied to identify their origin of medicinal plant species. In this review, we tried to overview the current research achievements for the development of suitable "DNA barcodes" regarding to the differentiation of medicinal plant species. Furthermore, more advanced techniques including amplification refractory mutation system (ARMS)-PCR, multiplex single base extension (MSBE), high-resolution melting (HRM) curve analyses are also discussed for their practical applications in the authentification of particular medicinal plant species.

Diversity Evaluation of Xylella fastidiosa from Infected Olive Trees in Apulia (Southern Italy)

  • Mang, Stefania M.;Frisullo, Salvatore;Elshafie, Hazem S.;Camele, Ippolito
    • The Plant Pathology Journal
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    • 제32권2호
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    • pp.102-111
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    • 2016
  • Olive culture is very important in the Mediterranean Basin. A severe outbreak of Olive Quick Decline Syndrome (OQDS) caused by Xylella fastidiosa infection was first noticed in 2013 on olive trees in the southern part of Apulia region (Lecce province, southern Italy). Studies were carried out for detection and diversity evaluation of the Apulian strain of Xylella fastidiosa. The presence of the pathogen in olive samples was detected by PCR amplifying the 16S rDNA, gyrase B subunit (gyrB) and HL hypothetical protein genes and single nucleotide polymorphisms (SNPs) assessment was performed to genotype X. fastidiosa. Twelve SNPs were recorded over gyrB and six SNPs were found for HL gene. Less variations were detected on 16S rDNA gene. Only gyrB and HL provided sufficient information for dividing the Apulian X. fastidiosa olive strains into subspecies. Using HL nucleotide sequences was possible to separate X. fastidiosa into subspecies pauca and fastidiosa. Whereas, nucleotide variation present on gyrB gene allowed separation of X. fastidiosa subsp. pauca from the other subspecies multiplex and fastidiosa. The X. fastidiosa strain from Apulia region was included into the subspecies pauca based on three genes phylogenetic analyses.

양식 강도다리, Platichthys stellatus Pallas에서 분리된 Streptococcus parauberis의 특성 (Isolation of Streptococcus parauberis from starry flounder, Platichthys stellatus Pallas)

  • 조미영;이재일;김명석;최희정;이덕찬;김진우
    • 한국어병학회지
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    • 제21권3호
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    • pp.209-217
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    • 2008
  • A disease with mass mortality characterized exophthalmia and abdominal distension by ascites occurred in farmed starry flounder, Platichthys stellatus in Pohang since October of 2007. Severe necrosis and infiltration of inflammatory cells were observed in tissue histopathological sections from liver, kidney, spleen and heart of the diseased fish. The catalase-negative and gram positive cocci with -hemolysis were isolated from moribund fish. Analysis of the API profiles and the multiplex PCR of the isolates showed that strains were identified as Streptococcus parauberis. In API profiles, S. parauberis isolated from starry flounder differed from those of olive flounder, Paralichthys olivaceus to the test of arginine dihydrolase, D-mannitol, D-sorbitol and α-lactose. And the strains, PH0710 and PH0711, were strongly lethal for starry flounder (LD50 ranging from 4.2×103~4.2×102 cfu/fish at 7 days after intraperitoneally inoculation). This is the first report on a S. parauberis infection in starry flounder.

Microbial Community Analysis of a Methane-Oxidizing Biofilm Using Ribosomal Tag Pyrosequencing

  • Kim, Tae-Gwan;Lee, Eun-Hee;Cho, Kyung-Suk
    • Journal of Microbiology and Biotechnology
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    • 제22권3호
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    • pp.360-370
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    • 2012
  • Current ecological knowledge of methanotrophic biofilms is incomplete, although they have been broadly studied in biotechnological processes. Four individual DNA samples were prepared from a methanotrophic biofilm, and a multiplex 16S rDNA pyrosequencing was performed. A complete library (before being de-multiplexed) contained 33,639 sequences (average length, 415 nt). Interestingly, methanotrophs were not dominant, only making up 23% of the community. Methylosinus, Methylomonas, and Methylosarcina were the dominant methanotrophs. Type II methanotrophs were more abundant than type I (56 vs. 44%), but less richer and diverse. Dominant non-methanotrophic genera included Hydrogenophaga, Flavobacterium, and Hyphomicrobium. The library was de-multiplexed into four libraries, with different sequencing efforts (3,915 - 20,133 sequences). Sorrenson abundance similarity results showed that the four libraries were almost identical (indices > 0.97), and phylogenetic comparisons using UniFrac test and P-test revealed the same results. It was demonstrated that the pyrosequencing was highly reproducible. These survey results can provide an insight into the management and/or manipulation of methanotrophic biofilms.

Characterization of Diarrheagenic Escherichia coli Isolated from Fresh Beef, Pork, and Chicken Meat in Korean Markets

  • Cho, Yong Sun;Koo, Min Seon;Jang, Hye Jin
    • 한국미생물·생명공학회지
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    • 제48권2호
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    • pp.121-128
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    • 2020
  • Diarrhea is a major public health concern associated with pathogenic Escherichia coli infections. Food-borne pathogenic E. coli can lead to large diarrheal outbreaks and hence, there is a need to estimate the frequency of pathogenic E. coli load in the various types of meat available in markets. In the present study, we classified and characterized diarrheagenic E. coli isolates collected from 399 raw meat samples from retail sources in Korea. Shiga toxin-producing E. coli (STEC) were detected in 11 (9.7%) samples, including nine strains (8.0%) in beef and two strains (1.8%) in chicken. The frequency of the detected virulence markers were as follows: astA, 28.3%; escV,18.6%; eaeA,17.7%; ent, 7.0%; EHEC-hly, 4.4%; stx1, 3.5%; and stx2, 3.5%. We did not observe any typical EPEC, EIEC, or ETEC virulence determinants in any of the samples. The STEC serotype O26 was detected in one sample, but no other serogroups (O91, O103, O128, O157, O145, O111, and O121) were found. Further research is needed to better understand the virulence mechanism of STEC serotypes, their ecology, and prevalence in animals, food, and the environment. These results will help improve risk assessment and predict the sources of food poisoning outbreaks.

Mating Types and Optimum Culture Conditions for Sexual StateFormation of Fusarium fujikuroi Isolates

  • Choi, Hyo-Won;Kim, Jung-Mi;Hong, Sung-Kee;Kim, Wan-Gyu;Chun, Se-Chul;Yu, Seung-Hun
    • Mycobiology
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    • 제37권4호
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    • pp.247-250
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    • 2009
  • Twenty-five isolates of Fusarium fujikuroi acquired from rice seeds and rice plants evidencing symptoms of Bakanae disease were evaluated to determine their mating types and characterize the formation of their sexual state. The mating types of the isolates were evaluated via multiplex PCR with the diagnostic primers of the mating-type (MAT) region: GFmat1a, GFmat1b, GFmat2c, and GFmat2d. Among the 25 isolates, 11 were identified as MAT-1 (male), and 14 as MAT-2 (female). Four MAT-1 isolates and three MAT-2 isolates were mated and cultured to evaluate the optimal culture conditions for the production of their sexual states. Among four tested media, 10% V8 juice agar proved optimal for the perithecial production of the isolates. The isolates also generated the largest numbers of perithecia when incubated at 23oC in alternating cycles of 12 hr fluorescent light and NUV fluorescent light and 12 hr darkness.