• Journal of Microbiology and Biotechnology
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• 제14권1호
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• pp.110-115
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• 2004

This study was undertaken to develop PCR primers for the simultaneous detection of Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans, using two species-specific reverse primers in combination with a single conserved forward primer. These primers target the variable and conserved regions of the 16S rDNA. The primer specificity was tested against (i) four F. nucleatum and three A. actinomycetemcomitans strains and (ii) seven representatives of the different species of oral bacteria. The primer sensitivity was determined by testing serial dilutions of the purified genomic DNA of F. nucleatum and A. actinomycetemcomitans. The data indicate that species-specific amplicons could be obtained for all the F. nucleatum and A. actinomycetemcomitans strains tested, which were not found in the seven other species. The multiplex PCR could detect as little as 4 fg of chromosomal DNA of F. nucleatum and A. actinomycetemcomitans simultaneously. These findings suggest that these PCR primers are highly sensitive and are suitable for applications in epidemiological studies, diagnosis, and monitoring F. nucleatum and A. actinomycetemcomitans after the treatment of periodontitis.

• Parasites, Hosts and Diseases
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• 제59권2호
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• pp.167-171
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• 2021

Haemonchosis remains a significant problem in small ruminants. In this study, the assay of recombinase polymerase amplification (RPA) combined with the lateral flow strip (LFS-RPA) was established for the rapid detection of Haemonchus contortus in goat feces. The assay used primers and a probe targeting a specific sequence in the ITS-2 gene. We compared the performance of the LFS-RPA assay to a PCR assay. The LFS-RPA had a detection limit of 10 fg DNA, which was 10 times less compared to the lowest detection limit obtained by PCR. Out of 24 goat fecal samples, LFS-RPA assay detected H. contortus DNA with 95.8% sensitivity, compared to PCR, 79.1% sensitivity. LFS-RPA assay did not detect DNA from other related helminth species and demonstrated an adequate tolerance to inhibitors present in the goat feces. Taken together, our results suggest that LFS-RPA assay had a high diagnostic accuracy for the rapid detection of H. contortus and merits further evaluation.

• 한국어류학회지
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• 제34권3호
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• pp.208-217
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• 2022

멸종위기어류 퉁사리 Liobagrus obesus를 대상으로 종 특이 프라이머 한 쌍과 프로브를 제작하여 하천수 시료에서 추출된 환경 DNA로부터 퉁사리를 검출할 수 있는 실시간 PCR 분석방법을 개발하고자 하였다. 퉁사리 종 특이 프라이머와 프로브는 미토콘드리아 DNA의 cytochrome b (cytb) 유전자 영역 내에서 국내에 서식하는 65종의 담수어류 간에 단일염기다형성 부위를 고려하여 비교한 후 제작하였다. 실시간 PCR 분석에서 제작한 프라이머 및 프로브는 국내에 서식하는 65종의 담수어류 gDNA를 이용한 특이성 검증 결과, 퉁사리 gDNA에서만 양성으로 나타나 높은 특이성을 보였다. 퉁사리 gDNA의 연속 희석 농도를 이용한 검출한계 분석에서는 0.2 pg까지 검출이 가능한 것으로 나타나 높은 감도를 보였다. 이후, 제작한 프라이머 및 프로브를 사용하여 금강 중·상류 유역의 8개 지점에서 확보한 하천수 시료를 대상으로 실시간 PCR 분석을 수행한 결과, 5개 지점에서 퉁사리의 cytb 유전자가 검출되었으며, 해당 검출 지점들은 현장 조사 당시에 퉁사리가 채집된 3개 지점을 모두 포함하였다. 따라서, 본 연구에서 개발한 퉁사리의 종 특이 프라이머와 프로브를 이용한 실시간 PCR 분석 방법은 하천수 채수로 확보한 환경 DNA로부터 퉁사리의 cytb 유전자를 검출할 수 있어 기존 서식지 모니터링과 더불어 잠재적인 신규 서식지 발굴에 활용될 수 있을 것으로 판단된다.

• Parasites, Hosts and Diseases
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• 제52권2호
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• pp.221-224
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• 2014

Larvae, nymphs, and adult stages of 3 species of ixodid ticks were collected by tick drag methods in Seoul during June-October 2013, and their infection status with severe fever with thrombocytopenia syndrome (SFTS) virus was examined using RT-PCR. During the period, 732 Haemaphysalis longicornis, 62 Haemaphysalis flava, and 2 Ixodes nipponensis specimens were collected. Among the specimens of H. longicornis, the number of female adults, male adults, nymphs, and larvae were 53, 11, 240, and 446, respectively. Ticks were grouped into 63 pools according to the collection site, species, and developmental stage, and assayed for SFTS virus. None of the pools of ticks were found to be positive for SFTS virus gene.

• 한국미생물·생명공학회지
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• 제31권2호
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• pp.205-209
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• 2003

Recently, we introduced a new method for rapid screening of bacterial species- or subspecies-specific DNA probes, named “inverted dot blot hybridization screening method”. We then applied this method to develop species- or strain- specific DNA probes for Prevotella intermedia and Prevotella nigrescens. In those studies, among 96 candidate DNA probes which were screened by the new method, 5 probes were confirmed as being putatively strain-specific : 3 probes for P. nigrescens 9336 (ATCC 33563), one for each p. intermedia ATCC 25611 and one for P. nigrescens G8-9K-3 (ATCC 49046). In the present study, we evaluated by Southern blot analysis a DNA probe Pi29-L, one of the 96 candidate probes described above, whether it is specific for the strain ATCC 25611 off. intermedia. Our data show that the probe Pi29-L is potentially P. intermedia ATCC 25611-specific, which can be useful for the detection and identification of the strain, particularly in maintenance of the strain.

• 한국응용과학기술학회지
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• 제10권1호
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• pp.1-8
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• 1993

The possibilities for HPLC analysis of lipids have been revolutionised by the availability of evaporative light-scattering detectors, with which the response is independent of the nature of the mobile phase and does not depend On the presence of specific chromophores in the lipids. It was thus possible to develop an HPLC procedure, involving ternary gradient elution, for separating all the lipid classes in animal tissues in a single step. Although reversed-phase HPLC has been widely used for the analysis of molecular species of lipids, sliver ion chromatography can be a valuable alternative. For example, a stable silver ion column for HPLC was developed which permitted resolution of molecular species of triacylglycerols, even from such complex samples as fish oils, again With light-scattering detection and gradient elution. The capacity for HPLC resolution of diastereomeric diacyl-sn-glycerol derivatives, prepared from triacylglycerols. has lead to a new simple method for stereospecific analysis of the latter.

• Parasites, Hosts and Diseases
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• 제48권4호
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• pp.343-346
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• 2010

The prevalence of the cercarial stage of an intestinal trematode, Haplorchis taichui, in thiarid snails (Gastropoda: Thiaridae) was investigated using light microscope and species-specific PCR procedures. A total of 988 snails were collected from Mae Taeng district, Chiang Mai province, northern Thailand, which comprised of 3 species; Melanoides tuberculata, Tarebia granifera, and Thiara scabra. The overall prevalence of pleurolophocercous cercariae was 21.7% as determined by the morphology. For genetic detection of H. taichui infection in snails, 2 primers Hapt_F (5'-GGCCAACGCAATCGTCATCC-3') and Hapt_R (5'-GCGTCGGGTTTCAGACATGG-3'), were used. The genomic DNA of H. taichui, which was used as a positive control, gave an amplification of the 256 bp fragment. The overall prevalence of H. taichui from specific PCR was 9.7%. The proportion of H. taichui among the pleurolophocercous cercariae in this study was 44.9%.

• The Plant Pathology Journal
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• 제23권4호
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• pp.266-271
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• 2007

The genomes of different allexiviruses were isolated and cloned from virus-infected garlic plants (Allium sativum), which were collected from farm fields in the southern provinces in Korea. The partial nucleotide sequences of the genomes from different allexiviruses were clearly identified in the virus-infected garlic plants. The cloned partial genomes of viruses in garlic plants showed a greater than 90% homology to previously identified allexiviruses and classified into species of GarV-A, -B, -C, -D, -E, and -X, demonstrating that species of allexivirus found in the other countries in the world are also widely distributed in the garlic plants in Korea.

• 융합보안논문지
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• 제5권2호
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• pp.65-74
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• 2005

최근 컴퓨터 네트워크의 발전에 따라 해킹사고도 급증하고 있으며 그 방법도 다양해지고 있다. 이러한 시기에 정보보호에 대한 관심이 높아지면서 많은 기업이 각종 보안시스템을 도입하고 있다. 그러나 해킹에 대응하기 위해서 대부분의 보안장비가 독자적인 이기종간의 기술을 적용해 제품간의 연동과 보안요소만으로 대처하기에는 많은 어려움이 있고, 이를 운용하는데 엄청난 조직, 장비, 인력 소요가 증대되고 있는 실정이다. 이러한 문제점을 해결하고자 이기종간 침입탐지 정보에 대한 보안요소를 통합하여 해킹으로부터 효율적으로 대응할 수 있는 웹 기반 관리 시스템을 설계 및 구현하였다.

• 한국임상수의학회지
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• 제35권6호
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• pp.273-275
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• 2018

A 6-month-old male cat was presented for investigation of depression, loss of appetite, dehydration, pale conjunctival mucous membrane, weight loss, fast heart and respiratory rates, nasal discharge and cough. Nasal swabs collected from the studied cat. As the results of bacterial culture with nasal swabs, it was suspected with Mycoplasma spp. Also, Mycoplasma species was detected by the PCR reaction with Mycoplasma genus primers. At species PCR assay, the specimens evaluated for the presence of M. felis, M. arginini, M. gateae, and Acholeplasma laidlawii and the result was visualization of bands from 238 bp in agarose gel 1.5% showing M. felis amplicons in samples. In conclusion, we detected M. felis in a cat with respiratory disease. PCR was able to detect successfully M. felis infection in cats.