• 한국음향학회지
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• 제26권1호
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• pp.42-47
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• 2007

본 논문에서는 probe DNA의 고정화 및 Probe DNA와 target DNA의 혼성화 반응을 감지할 수 있는 DNA 측정용 고주파 SAW 센서의 주파수 증대에 따른 감도향상에 대해 연구하였다. 센서는 $36^{\circ}$ YX $LiTaO_3$ 압전 단결정 기판위에 Au 박막이 증착된 측정채널 (sensing channel)과 기준채널 (reference channel)로 구성되며 200MHz에서 발진되는 이중 지연선 형태로 제작되었다. 또한 SAW 센서의 감지 미케니즘의 최적화를 위해 SAW 센서의 Au 지연선상의 Probe DNA의 최적 고정화 반응농도와 target DNA의 최적 혼성화 반응농도를 결정하였으며, 디지털 시린지 펌프시스템을 구성하여 실험자에 따른 오차를 최소화하였다. 측정채널의 Au 박막 지연선상에 probe DNA를 고정화시킨 후 target DNA를 주입하면, DNA의 혼성화 반응이 일어나며 Au 지연선상의 질량이 변하게 된다. 따라서 질량하중 효과에 대한 센서의 주파수 변화를 측정하였다. 개발된 센서는 최대 0.066ng/ml/Hz의 민감도를 가지며 질량하중 효과에 대한 안정적인 주파수 변화를 나타내었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.235-236
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• 2001

유전자 분석 기술은 biomedical analysis를 위해 일반적으로 고체 상에 고정화된 DNA 분자를 이용한다. 이 센서의 검출능력은 주로 capture probe의 서열뿐만 아니라 oligonucleotide의 고체 상에 고정화 방법에 달려있다. 본 연구에서는 glass 표면에 DNA 분자를 고정화시키는 방법과 PCR product를 정제하지 않고 직접 검출할 수 있는 DNA probe assay 방법을 개발하였다.

• 한국식물병리학회지
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• 제14권2호
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• pp.164-170
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• 1998

This study was carried out to develop DNA probe for specific detection of Erwinia carotovora subsp. carotovora. Universal rice primer (URP, 20 mer) developed from repetitive sequence of rice was applied for producing PCR DNA fingerprints of Erwinis spp. In E. carotovora subsp. carotovora strains, primer URP2F amplyfied polymorphic bands which are distinguisable from other Erwinia spp. A PCR band of 0.6 kb selected from PCr polymorphic bands of E. carotovora subsp. carotovora strains was cloned and evaluated as a diagnostic DNA probe. Among 28 bacterial strains including 22 Erwinia spp, the probe (pECC2F) only hybridized to total DNAs from e. carotovora subsp. carotovora strains and E. carotovora subsp. wasabiae, but sizes of hybridized bands were different between these subspecies, 10.0 kb and 3.5 kb respectively. In dot blot assays using probe pECC2F, as few as 103 colony forming units (CFU) of E. carotovora subsp. carotovora could be detected in a suspension containing about 1$\times$103 CFU of soil bacteria.

• Asian-Australasian Journal of Animal Sciences
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• 제15권8호
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• pp.1071-1075
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• 2002

In order to identify and develop the specific DNA marker for the identification of Hanwoo (Korean Cattle) from other breeds, a specific DNA marker of 519 bp was identified and sequenced from polymorphic analysis using RAPD-PCR for 6 cattle breeds. Two different repetitive sequences, $(AAC)_5$ and $(GAAGA)_2$, were selected and designed to use specific probe to develop a DNA marker for Hanwoo specific. When the $(AAC)_5$ probe was applied, the 10 kb specific DNA marker showed in the DNA fingerprinting from 237 of 281 Hanwoo individuals. This novel Hanwoo specific DNA probe is useful to perform the marker-assisted selection for screening Hanwoo purity as an unique genetic source.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.711-712
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• 2000

DNA probe assay comprising a microwell as' solid matrix for the immobilization of streptavidin (SA) and an oligonucleotide with covalently bound fluorecein as detection probe was developed. The insolubilized SA captured the biotinylated DNA product of polymerase chain reaction (PCR), and the product was denatured under a basic condition. The remaining single-stranded DNA on the solid surface was hybridized with the probe for signal generation that was performed based on enzyme-linked immuno -reactions.

• Journal of Periodontal and Implant Science
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• 제32권2호
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• pp.281-290
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• 2002

The purpose of this study is to develop species-specific DNA probe for detection and identification of Prevotella intermedia (P. intermedia) G8-9K-3. This study procedure includes (1) whole-genomic DNA extraction of P. intermedia G8-9K-3 (2) construction of the genomic DNA library, (3) screening of strain-specific DNA probe by reverse dot hybridization, (4) confirmation of strain-specific DNA probe by Southern blot hybridization, (5) determination of nucleotide sequences of strain-specific DNA probe. Twenty-eight recombinant plasmids containing Hind III-digested DNA fragments of P. intermedia G8-9K-3 were obtained. Reverse Dot Hybridization and Southern blot analysis data showed that one of them, Pig3, could be P. intermedia G8-9K-3-specific DNA probe. This datum indicates that this Pig3 DNA probe could be useful in detection and identification of the P. intermedia G8-9K-3 strain.

• 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.

• 한국식품과학회지
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• 제35권3호
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• pp.470-474
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• 2003

식중독은 세균에 의한 발병이 대부분이다. 따라서 식품에서 식중독 원인균을 신속하게 탐색하게 식중독으로부터의 되면 피해를 줄일 수 있을 것이다. 고전적인 식중독 원인균 탐색은 증균, 선택적 배지를 이용한 isolation, 생화학적 특징을 활용하는 분석이 있으나 많은 시간이 소요되는 단점을 갖고 있었다. 본 연구는 16S rRNA gene(16S rDNA)로부터 얻은 DNA 염기 서열을 이용 식중독 원인균의 특이적 oligonucleotide probe을 제작 reverse dot blot hybridization과 PCR 방법을 이용하여 고전적인 방법보다 빠른 시간 내에 식품에서 원인균을 탐색 할 수 있었다. 우유를 인공적으로 본 연구에서 사용한 균주로 오염시킨 후 DNA를 추출하여 PCR 증폭산물과 oligonucleotide probe를 hybridization 시킨 결과 oligonucleotide probe를 hybridization 시킨 결과 oligonucleotide probe가 위치한 곳에서 발색 반응이 나타났다. 본 연구에서 본 연구를 통해 DNA microchip으로 활용 짧은 시간 내에 많은 종류의 식중독 원인균을 탐색 할 수 있는 가능성을 확인하였다.

• BMB Reports
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• 제37권3호
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• pp.356-361
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• 2004

To develop a simplified method that can rapidly prepare DNA microarray probes in a massive scale, a lambda phage genomic DNA-fragments library was constructed for the microarray-probes collection. Four methods of DNA band recovery from the first PCR products were tested and compared. The DNA microarray probes were collected by a novel method of nested PCR that was mediated by gel isolation of the first PCR products. This method was named GIN-PCR. The probes that were prepared by this GIN-PCR technique were used as subjects to fabricate a DNA microarray. The results showed that a wooden toothpick was superior to the other 3 methods, since this technique can steadily transfer the DNA bands as the template of the second PCR after the first PCR. A group of probes were successfully collected and DNA microarrays were constructed using these probes. Hybridization results demonstrated that this technique of DNA recovery and probe preparation was rapid, efficient, and effective. We developed a cost-effective and less labor-intensive method for DNA microarray probe preparation by nested PCR that is mediated by wooden toothpick transfer of the DNA bands in the gel after electrophoresis.

• Restorative Dentistry and Endodontics
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• 제21권1호
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• pp.1-18
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• 1996

There are many advantages when using IIF and DNA probe methods over anaerobic culture method in that they are time-and effort-saving, more precise and more sensitive. Furthermore, in IIF and DNA probe methods, the detection is possible only with small amount of bacteria, the quantitative analysis is possible, and the cell viability is not necessary. The purpose of this study is to observe the incidence of P.endodontalis by carrying out anaerobic culture, IIF and colony lift using DNA probe method respectively, and to compare these 3 methods in terms of effectiveness and sensitivity in order to identify the most effective detection method. 30 teeth with at least one clinical symptoms, with single canal, and with pulp necrosis were sampled. For sampling bacteria, access cavity was prepared after disinfecting tooth and its surroundings. Then the paper point was inserted up to the periapical area, leave there for a while, and finally it was placed into PRAS Ringer's sol. and PBS sol. In anaerobic culture method, P.endodontalis was identified by biochemical tests after subculturing black and brown colonies which were produced after 7 days of incubation on BAP and Brucella BAP in anaerobic chamber. To identify P.endodontalis in IIF method, species-specific polyclonal rabbit-antisera of P.endodontalis(ATCC 35406) was reacted with sampled PBS sol. dispensed onto glass slide, and then P.endodontalis was examined by phase contrast microscopy after incubating with Goat anti-rabbit lgG conjugated to Fluorescein isothiocyanate. For colony lift using DNA probe method, membranes were laid over colonies on the surface of BAP and were hybridized with cloned DNA probe of P.endodontalis. The existence of P.endodontalis was then identified by the methods of chemiluminescent detection and color metric detection. Black colony was found in 11 teeth out of 30 teeth and P.endodontalis was detected in 6 teeth (20 %) by anaerobic culture method, 16 teeth (53 %) by IIF method, and 7 teeth (23 %) by DNA probe method. IIF method is significantly better in detecting P.endodontalis than DNA probe method and anaerobic culture method. There was no significant differences between DNA probe method and anaerobic culture method. There was significant correlation between the formation of black colony and the existence of P.endodontalis. The probability of detecting P.endodontalis when black colony being present is 2.89 times higher than when not being present. There was significant relationship between the foul odor of clinical symptoms and P.endodontalis. The sensitivity of existing P.endodontalis when foul odor being present was 93.75 %, while the specificity of not existing P.endodontalis when foul odor not being present was 28.57 %. These results suggested that the probes of P.endodontalis will be used to decide the method and prognosis in endodontic treatments.