• 대한수의학회지
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• 제40권1호
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• pp.56-62
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• 2000

Pasteurella multocida is kind of commensal bacteria in the upper respiratory tract of pigs. It is classified toxigenic and nontoxigenic strains based on the production of dermonecrotic toxin. Toxigenic strain is most associated with atrophic rhinitis which brings great economical loss in swine industry. However, toxigenic and nontoxigenic strains do not differ by diagnostic biochemical reaction or morphology. One of recently developed techniques, PCR detects the toxigenic P multocida. Amplification of an 846-nucleotide fragment of toxA gene was developed. The fragment amplified by PCR was detected in P multocida type D not type A. The PCR amplification was as sensitive as it could detect 1 pg of P multocida DNA. We compared the result of the PCR with the enzyme linked immunosorbent assay (ELISA) in a test for 40 swine nasal swabs. All of these isolates were toxin negative based on the ELISA while 2 isolates were detected in the PCR technique. in addition to accuracy, as required for rapid detection from contaminated nasal swabs, toxigenic P multocida was recovered efficiently from contaminated culture without inhibition of the PCR. The results show that the PCR detection of toxigenic P multocida directly form nasal swabs are feasible.

• Fisheries and Aquatic Sciences
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• 제5권1호
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• pp.54-58
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• 2002

The polymerase chain reaction (PCR) amplification technique was used for comparison of Listeria monocytogenes serotypes. PCR primers for the fragment of invasion-associated protein (iap) gene were highly specific for all the serotypes of L. monocytogenes. Other Listeria spp., such as Listeria ivanovii and Listeria innocua were not produced the PCR fragments by above primer set. The nucleotide sequences of PCR products showed high homologies in comparison of all the isolated serotypes except unknown type II-2. The deduced amino acid sequences of the PCR products also showed similar to one another. The various region of the PCR products, called a Thr-Asn repeat region was presented. All of isolated L. monocytogenes serotypes possessed 16 to 20 Thr-Asn repeats.

• Preventive Nutrition and Food Science
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• 제3권2호
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• pp.198-202
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• 1998

The arbitrary primed polymerase chain reaction (AP-PCR) has been used to detect the genetic alternations in the related species. Simple and reproducible fingerprints of complex genomes can be generated using single arbitrary chosen primers and the PCR. The technique was applied to the Oryza species and characterized the relationship among three cultivars of rice species based on theresult of genomic DNA fingerprints. The results indicated that the polymorphism revealed in rice strains and the differences in the PCR product pattern could be represented for each strainis. There was many variationsin the PCR product pattern between cv. Dongin(japonica type)and cv.Hyangdo (indica type), and our chosen AP-primers can ge as markers for strain identification and verfication.

• Asian Pacific Journal of Cancer Prevention
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• 제15권24호
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• pp.10647-10652
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• 2015

Background: The aim of our study was to establish COLD-PCR combined with an unlabeled-probe HRM approach for detecting KRAS codon 12 and 13 mutations in plasma-circulating DNA of pancreatic adenocarcinoma (PA) cases as a novel and effective diagnostic technique. Materials and Methods: We tested the sensitivity and specificity of this approach with dilutions of known mutated cell lines. We screened 36 plasma-circulating DNA samples, 24 from the disease control group and 25 of a healthy group, to be subsequently sequenced to confirm mutations. Simultaneously, we tested the specimens using conventional PCR followed by HRM and then used target-DNA cloning and sequencing for verification. The ROC and respective AUC were calculated for KRAS mutations and/or serum CA 19-9. Results: It was found that the sensitivity of Sanger reached 0.5% with COLD-PCR, whereas that obtained after conventional PCR did 20%; that of COLD-PCR based on unlabeled-probe HRM, 0.1%. KRAS mutations were identified in 26 of 36 PA cases (72.2%), while none were detected in the disease control and/or healthy group. KRAS mutations were identified both in 26 PA tissues and plasma samples. The AUC of COLD-PCR based unlabeled probe HRM turned out to be 0.861, which when combined with CA 19-9 increased to 0.934. Conclusions: It was concluded that COLD-PCR with unlabeled-probe HRM can be a sensitive and accurate screening technique to detect KRAS codon 12 and 13 mutations in plasma-circulating DNA for diagnosing and treating PA.

• 한국식품과학회지
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• 제33권5호
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• pp.521-524
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• 2001

본 연구는 유전자재조합 기술에 의해 개발된 glyphosate에 내성을 가지고 있는 콩(GTS)의 모니터링을 위하여 PCR을 이용한 검출 방법에 대한 실험을 수행하였다. Glyphosate에 내성이 있는 콩에 삽입된 유전자와 표준대조 유전자인 lectin과 ferritin 유전자를 근거로 제작된 primer와 CTAB 방법으로 추출된 콩의 DNA를 template로 이용하여 PCR을 수행하였다. GTS의 검출을 위한 제작된 primer들은 GTS와 특이적으로 반응하여 증폭된 PCR 산물을 생성하였으나, non-GTS와는 PCR 산물을 생성하지 못했다. 증폭된 염기서열 분석을 통하여 GTS에 특이적인 것을확인하였으며, 약 0.05%가 포함되어 있는 GTS까지 검출이 가능함을 보였다.

• 대한임상검사과학회지
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• 제36권1호
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• pp.1-6
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• 2004

The enterobacterial repetitive intergenic consensus (ERIC)-PCR is a recently described DNA fingerprinting technique based on amplification of repetitive element distributed in bacteria. We applied of ERIC-PCR to clinical isolates of Vibrio parahaemolyticus and other bacteria associated diarrhea. Twenty isolates of V. parahaemolyticus were used for intragenic genotyping, which were isolated from 2001 to 2002 in Chungbuk National University hospital. For interspecies genotyping, V. vulnificus, V. alginolyticus, V. parahaemolyticus, Escherichia coli, Salmonella and Shigella spp. were used. The genotyping were analyzed by ERIC-PCR. The genotyping of V. parahaemolyticus were grouped two major pattern (A, B) and were subdivided into 10 subtypes (A1, A2, B1-B8) by ERIC-PCR. These method distinctly differentiated bacterial species associated diarrhea. Those results show that ERIC-PCR can be reliable and efficient method for genotyping of V. parahaemolyticus and bacteria associated diarrhea.

• 미생물학회지
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• 제37권1호
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• pp.56-60
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• 2001

탄저균의 분자적 다양성 분석은 다양한 DNA표지의 부족으로 쉬운 일이 아니어서, 본 연구에서는 random amplified polymorphic DNA (RAPD)-PCR을 이용하여 Bacillus 속으로부터 탄저균을 구별할 수 있는 새로운 DNA 표지를 개발하고자 하였다. RAPD-PCR을 이용한 분석은 다양한 Bacillus 종으로부터 탄저균을 동정할 수 있었으며, 아울러 Bacillus 종 사이에서 확실한 유전적인 변이를 확인할 수 있었다. 이러한 분석은 간단, 신속하고, 그리고 정확하게 탄저균을 진단하는데 활용할 수 있다고 본다.

• 한국습지학회지
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• 제14권3호
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• pp.419-428
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• 2012

식물플랑크톤의 동정은 숙련된 전문가에게도 어려운 과제이다. 별 특징없는 외형과 다양한 크기와 종은 형태학적으로 구분하기에 어려움이 있다. 본 연구에서는 미생물 군집의 다양성을 분석하는데 효과적인 fingerprinting 기법인 PCR-DGGE 방법을 사용하여 이런 형태학적 동정의 제한점을 보완하고자 하는데 목적이 있다. 5곳의 호수 샘플로부터 2008년 8월 총 46개의 band를 찾을 수 있었고, 2008년 11월 총 26개 band를 찾을 수 있었다. 이 fingerprint 결과는 각각 다른 샘플링 장소를 비교하는데 용이하였다. 본 연구에서 PCR-DGGE 방법은 북한강 호수들의 플랑크톤 군집의 다양성을 파악하는데 사용되었고, 이 DGGE 기법이 플랑크톤의 동정기법으로써의 가능성을 검토해보았다.

• 한국식물병리학회지
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• 제13권4호
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• pp.205-209
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• 1997

PSTVd 를 접종시킨 감자의 Superior 품종으로부터 PSTVd RNA를 분리하여 역전사효소와 PSTVd genome 중 356 uncleotides를 증폭할 수 있는 PSTVd 특이적 primer 쌍을 사용하여 RT-PCR를 수행한 결과 356 nuclcotides의 DNA 절편이 증폭되었고 이 절편을 sequencing인 결과 PSTVd 의 유전자임을 확인하였다. 감염된 잎과 괴경에수 모두 RT-PCR 기법을 이용한 PSTVd 의 검정이 가능하였고 특히 RT 반응시 downstream primer만을 이용할 때보다 downstream과 upstream primer를 동시에 사용할 때가 PSTVd 의 검정에 더 효과적이었다.

• Journal of Microbiology and Biotechnology
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• 제31권12호
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• pp.1709-1715
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• 2021

Outbreaks of food poisoning due to the consumption of norovirus-contaminated shellfish continue to occur. Male-specific (F+) coliphage has been suggested as an indicator of viral species due to the association with animal and human wastes. Here, we compared two methods, the double agar overlay and the quantitative real-time PCR (RT-PCR)-based method, for evaluating the applicability of F+ coliphage-based detection technique in microbial contamination tracking of shellfish samples. The RT-PCR-based method showed 1.6-39 times higher coliphage PFU values from spiked shellfish samples, in relation to the double agar overlay method. These differences indicated that the RT-PCR-based technique can detect both intact viruses and non-particle-protected viral DNA/RNA, suggesting that the RT-PCR based method could be a more efficient tool for tracking microbial contamination in shellfish. However, the virome information on F+ coliphage-contaminated oyster samples revealed that the high specificity of the RT-PCR- based method has a limitation in microbial contamination tracking due to the genomic diversity of F+ coliphages. Further research on the development of appropriate primer sets for microbial contamination tracking is therefore necessary. This study provides preliminary insight that should be examined in the search for suitable microbial contamination tracking methods to control the sanitation of shellfish and related seawater.