• 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.

• 대한수의학회지
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• 제37권2호
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• pp.367-374
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• 1997

Bovine rotaviruses(A, 288, 55086 strains) isolated from fecal samples in Korea were propagated onto MA104 cells and were confirmed tentatively as G6, G8, and G10, respectively, by RFLP analysis. Full-length VP7 gene of these isolates was amplified by reverse transcriptase polymerase chain reaction(RT-PCR) using VP7 specific primers and cloned into TA vector. Nucleotide and deduced amino acid sequences of VP7 genes of the isolates were determined and compared with those of bovine rotavirus reference strains(NCDV; G6, UK; G6, Cody I-801; G8 and B223; G10). A, 288 and 55086 isolates showed high degree of nucleotide sequence homology with NCDV and UK(93% and 94%), Cody I-801(86%) and B223(97%), respectively, However, they showed 71~74% of nucleotide sequence homlogy with bovine rotavirus reference strains which belong to different serotypes. From the results of deduced amino acid sequence homology analysis, three isolates showed 94~96% of homology with the same serotype reference strains but 80~84% of homology with the different serotype reference strains. Three bovine rotavirus isolates, A, 288 and 55086 strains, were confirmed as G6, G8, and G10, respectively, by nucleotide and deduced amino acid sequence analysis.

• Journal of Microbiology
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• 제44권1호
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• pp.106-112
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• 2006

In this study, the genetic diversities of multi-resistant Salmonella typhimurium (ST) isolates were analyzed via the application of both pulsed field gel electrophoresis (PFGE) and Polymerase chain reaction (PCR) analysis methods, using 6 kinds of primers (REP, ERIC, SERE, BOX, P-1254 and OPB-17). And their discriminative abilities (DA) were also compared in order to determine the most effective and reliable analysis method. 118 S. typhimurium isolates, cultured from diverse animals and human patients in Korea beginning in 1993, were analyzed and subjected to a comparison of Simpson's index of diversity (SID), using both PFGE and PCR methods. PFGE by XbaI enzyme digestion allowed for discrimination into 9 pulsotypes, with high SID values (0.991) on the genomic DNA level. This shows that PFGE is a very discriminative genotypic tool, and also that multiple clones of S. typhimurium isolates had existed in domestic animals and humans in Korea since 1993. However, we could ultimately not to trace the definitive sources or animal reservoirs of specific S. typhimurium isolates examined in this study. Depending on the SID values, the combined method (7 kinds of method) was found to be the most discriminative method, followed by (in order) SERE-PCR, REP-PCR, ERIC-PCR, PFGE & OPB-17 (RAPD), P-1254 (RAPD), and BOX-PCR at the $80\%$ clone cut-off value. This finding suggests that the REP-PCR method (which utilizes 4 primer types) may be an alternative tool to PFGE for the genotyping of S. typhimurium isolates, with comparable cost, time, and labor requirement. The establishment of a highly reliable and discriminatory method for epidemiologic analysis is considered necessary in order for researchers to trace the sources of specific pathogens and, consequently, to control and prevent the spread of epidemic S. typhimurium isolates to humans.

• 한국식품과학회지
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• 제31권1호
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• pp.183-188
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• 1999

국내 식육중 Y. enterocolitica의 분포 조사 및 분리균의 특성을 조사하기 위하여 4월부터 11월사이에 상, 하반기로 나누어 소고기, 돼지고기, 닭고기 463건을 서울, 대전, 광주, 부산에서 구입하여 실험하였다. 전체 식육중 17.5% (81건)에서 Y. enterocolitica가 분리되었으며 닭고기(26.0%)가 가장 많이 오염되어 있었다. 분리율은 계절에 따라 차이를 보였는데 하반기에 수거한 돼지고기와 닭고기에서 상반기보다 2배이상 높은 균 분리율을 보였다. 항생제 감수성 실험결과 대부분의 분리균은 carbenicillin, ampicillin, erythromycin, penicillin, bacitracin에 대해 내성을 나타내는 반면 polymyxin B, kanamycin, trimethoprim/sulfamethoxazole, ciprofloxacin, tetracycline, nalidixic acid, gentamicin등에 감수성을 나타내었다. 분리된 Y. enterocolitica의 혈청형은 serotype O:5가 주종이었고 일본 및 유럽 등지에서 많이 분리되어지는 O:9는 분리되지 않았다. PCR 실험결과 81개의 분리균주중 8균주(10%)가 병원성을 갖고있는 것으로 밝혀졌는데 그 중 6균주가 돼지고기에서 발견되었다. 그리고 돼지고기 분리균주중 병원성이 없는 것으로 널리 알려져 있는 serotype O:5가 PCR 양성반응으로 병원성을 보여 이전 연구들과 상반된 결과를 나타낸 것에 대해서는 더 연구해야할 과제로 남아 있다.

• 한국식품과학회지
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• 제31권5호
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• pp.1324-1329
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• 1999

국내 유통중인 냉동만두와 피자 중 L. monocytogenes의 분포와 분리균의 특성을 조사하기 위하여 1998년 8월부터 11월에 걸쳐 시료 72개를 구입하여 실험하였다. 전체 냉동식품 중 9.7%(7개)에서 L. monocytogenes가 분리되었으며 냉동만두 중 11.1%,피자 중 5.6%가 오염되어 있었다. USDA와 FDA방법. 그리고 modified cold enrichment 방법 중 USDA방법이 가장 L. monocytogenes의 분리율이 높았으며 분리된 L. monocytogenes 혈청형은 4였다. PCR실험결과 CAMP test와 API Listeria kit를 사용하여 분리된 균주가 L. monocytogenes임이 확인되었다. USDA방법과 PCR을 이용하여 L. monocytogenes를 분리하고 확인하는데 3-4일 정도의 시간이 소요되어 시간을 단축시킬 수 있는 분리 및 확인방법의 개발이 필요하리라 사료된다. 분리된 L. monocytogenes의 내열성을 조사한 결과 tryptic soy broth에서 D값이 $60^{\circ}C$에서 49.2초, $65^{\circ}C$에서 8.8초였으며 냉동식품의 안전한 사용을 위하여 L. monocytogenes의 분리율이 높은 식품에서 D값의 측정이 필요하다고 사료된다.