• 미생물학회지
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• 제47권2호
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• pp.103-109
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• 2011

Pectobacterium carotovorum subsp. carotovorum (PCC)는 세균성 무름병균으로 주로 감자, 양배추 등의 식물에서 질병을 일으킨다. 본 연구에서는 현장에서 신속하게 진단하기 위해 loop-mediated isothermal amplification법을 이용하여 1시간 내에 등온에서 검출 가능한 진단법을 개발하였으며, 이를 PCC-LAMP법이라 명명하였다. PCC의 lytic murein transglycolase 유전자를 특이적으로 증폭시키는 4개의 프라이머를 제작하였으며 최적 온도가 $61^{\circ}C$임을 확인하고 최적 조건을 확립하였다. 최적 조건을 바탕으로 4개의 프라이머가 $1{\times}10^3$ copies까지 검출하는 민감성을 확인할 수 있었다. 본 연구에서 개발된 PCC-LAMP법은 특이성 검사를 통해 PCC만이 특이적으로 검출됨을 확인하였으며, 이는 실제 시료에서도 적용 가능함을 확인하였다. PCC-LAMP법을 통하여 PCC를 신속하고 정확하게 검출함으로써 현장에서 유용하게 적용될 수 있을 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제20권1호
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• pp.193-201
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• 2010

A specific and rapid real-time PCR assay for detecting Ralstonia solanacearum in horticultural soil and plant tissues was developed in this study. The specific primers RSF/RSR were designed based on the upstream region of the UDP-3-O-acyl-GlcNAc deacetylase gene from R. solanacearum, and a PCR product of 159 bp was amplified specifically from 28 strains of R. solanacearum, which represent all genetically diverse AluI types and all 6 biovars, but not from any other nontarget species. The detection limit of $10^2\;CFU/g$ tomato stem and horticultural soil was achieved in this real-time PCR assay. The high sensitivity and specificity observed with field samples as well as with artificially infected samples suggested that this method might be a useful tool for detection and quantification of R. solanacearum in precise forecast and diagnosis.

• 한국미생물·생명공학회지
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• 제37권3호
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• pp.238-242
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• 2009

분자생물학적 방법에 의한 김치유산균 연구의 진행으로 Leuconostoc, Lactobacillus, Weissella 속 유산균의 김치발효 과정 중 생육에 대한 새로운 결과들이 도출되었지만, Pediococcus속의 생육에 대한 새로운 결과는 거의 보고되지 않았다. 본 연구에서는 김치에 존재하는 Pediococcus 속 유산균의 존재에 대한 재조명을 위하여 ampicillin (A)의 첨가로 pediococci의 선발 유효성이 향상된 것으로 보고된 pediococci selective medium(PSM)+A를 이용하여 pediococci 선택적 검출의 유효성을 검토한 결과, 기존에 보고된 결과와는 달리 Pediococcus pentosaceus 외에도 leuconostocs, Lactobacillus casei, Lactobacillus curvatus, Oenococcus oeni, Streptococcus thermophilus에 대한 생육저해가 발생하지 않았다. PSM+A 한천배지를 이용하여 김치에 생육하는 미생물을 검출한 경우, leuconostocs, P. pentosaceus, Weissella koreensis, Lb. curvatus, Lactobacillus brevis, Lactobacillus sakei가 생육하여 김치와 같은 발효침채류의 발효에 관여하는 pediococci의 선택적 선발에는 PSM+A가 유효하지 않은 것으로 확인되었다.

• Fisheries and Aquatic Sciences
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• 제7권3호
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• pp.148-162
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• 2004

The monitoring activities at the National Fisheries Research and Development Institute (NFRDI) in Korea have been extended to include all the coastal waters of Korea after the outbreak of Cochlodinium polykrikoides blooms in 1995. We used several alternative methods including climatological analysis, spectral and optical methods which may offer potential detection of the major species of red tide in Korean waters. In the climatological analysis, NOAA, SeaWiFS, OCM satellite data was chosen using the known C. polykrikoides red tide bloom data and the area was mapped by helicopter reconnaissance and ground observation. The relationship between the distribution of sea surface temperature to C. polykrikoides bloom areas was studied. The anomalies of SeaWiFS chlorophyll a imageries against the imageries of non-occurring red tide for August, 2001 showed where the C. polykrikoides occurred. The anomalies of chlorophyll a concentrations from the satellite data during red tide outbreaks showed a similar distribution of C. polykrikoides in the red tide in August, 2001. The distribution between differences in sea surface temperatures during the day and at night also showed a possibility for red tide detection. We used a corrected vegetation index (CVI) to detect floating vegetation and submerged vegetation containing algal blooms. The results of from the optical absorption of C. polykrikoides in the ultraviolet band (340 nm) showed that if we use the optical characteristics from each red tide, we will be able to establish the feasibility of red tide detection.

• The Plant Pathology Journal
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• 제15권6호
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• pp.335-339
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• 1999

Citrus tristeza virus(CTV), an aphid-borne closterovirus, is one of the most destructive pathogens of citrus. It has caused rapid decline in growth, stem pitting and death in citrus trees. A reverse transcriptase polymerase chain reaction (RT-PCR) assay was developed for detection of CTV and investigation of the CTV infection status of citrus and its related cultivars in Cheju island. For RT-PCR based CTV detection, primers were designed to amplify 670bp of coat protein gene. A screening test for CTV in citrus cultivars was conducted from March to July in 1999. Seventy individual citrus trees representing 9 species of 3 genera were tested. The infection rates of CTV for leaves from the years or older trees of late maturing citrus varieties such as Yuzu (C. junos Sieb. ex Tanaka), Navel orange (C.sinensis Osbeck), Kiyomitanger (C. unshiu x C. sinensis), and Shiranuhi ((C. unshiu x C. sinensis) x C. reticulata) were 100%, 80%, 60%, and 60% respectively. The CTV infection rates in Early satsuma mandarins such as 'Miyagawa Early' Satsuma mandarins (C. unshiu Marc. var. Miyagawa) and 'Okitsu Early' Satsuma mandarins (C. unshiu Marc. var. Okitsu) were 100%, and 60%, respectively. CTV was not detected in Cheju native Dangyooja (C. unshiu Marc. var. Osbeck), Trifoliate orange (Poncirus trifoliata) and Kumquat (Fortunella margarita Swingle). In conclusion, RT-PCR assay can be successfully applied to the detection of CTV in citrus trees.

• Journal of Microbiology and Biotechnology
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• 제23권2호
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• pp.246-250
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• 2013

In this study, a loop-mediated isothermal amplification (LAMP) method to rapidly detect Staphylococcus aureus strains was developed and evaluated by extensively applying a large number of S. aureus isolates from clinical and food samples. Six primers were specially designed for recognizing eight distinct sequences on the species-specific femA gene of S. aureus. The detection limits were 100 fg DNA/tube and $10^4$ CFU/ml. The LAMP assay was applied to 432 S. aureus strains isolated from 118 clinical and 314 food samples. Total detection rates for the LAMP and polymerase chain reaction assays were 98.4% (306/311) and 89.4% (278/311), respectively.

• 한국자원식물학회지
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• 제23권3호
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• pp.233-241
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• 2010

In vitro elimination of Sweet potato leaf curl virus (SPLCV) from infected sweet potato is difficult due to low number of virus-free plants obtained from meristem tip culture and long growth period required for the virus detection. In this study, efficient production of the SPLCV-free sweet potato by in vitro therapy coupled with a PCR assay for virus detection was investigated. Infected shoots cultured on Murashige and Skoog medium were treated at three different temperatures for 7 weeks followed by meristem tip culture on the medium with or without ribavirin at 50 mg/L. The regenerated plantlets were tested for virus infection by a PCR assay. The results showed that the both heat- and cold-treatments, and addition of the ribavirin did not have significant effect on efficiency of the virus elimination. The meristem size, however, greatly affected the survival rate. Meristems sized over 0.4 mm survived better than smaller ones (0.2-0.3 mm). The PCR assay was approved to be a rapid, sensitive and reliable for the SPLCV detection in regenerated plantlets. Therefore, combination of cultivating meristem tips sized 0.4-0.5 mm on the medium at $22^{\circ}C$ without ribavirin and detection of SPLCV in the regenerated plantlets by the PCR assay was an efficient system for the SPLCV elimination from infected sweet potato.

• Food Science and Biotechnology
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• 제17권4호
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• pp.761-765
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• 2008

Bacillus cereus causes two different types of food poisoning syndromes: diarrhea and emesis. The diarrheal syndrome is attributed to various enterotoxins, including nonhemolytic enterotoxin, hemolytic enterotoxin, and enterotoxin-T, whereas the emetic syndrome is caused by the dodecadepsipeptide toxin cereulide. A multiplex polymerase chain reaction (PCR) assay was developed to rapidly detect and identify B. cereus strains. Three primer pairs specific to regions within genes encoding nonhemolytic enterotoxin (nheA), molecular chaperonin (groEL), and cereulide synthetase (ces) were used to identify and differentiate between the enterotoxin-producing and emetic toxin-producing B. cereus strains. The cereulide-producing emetic B. cereus showed 3 PCR products of 325, 405, and 685 bp for the groEL, ces, and nheA genes, respectively, whereas the enterotoxin-producing B. cereus showed 2 PCR products without a ces gene specific DNA fragment. Specific amplifications and differentiations by multiplex PCR assay were obtained using 62 B. cereus strains and 13 strains' of other bacterial species. The detection limit of this assay for enterotoxin-producing strain and emetic toxin-producing strain from pure cultures were $2.4{\times}10^1$ and $6.0{\times}10^2\;CFU/tube$, respectively. These results suggest that our multiplex PCR method may be useful for the rapid detection and differentiation of B. cereus strains in foods.

• The Plant Pathology Journal
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• 제38권6호
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• pp.629-636
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• 2022

Colletotrichum species is known as the major causal pathogen of red pepper anthracnose in Korea and various groups of fungicides are registered for the management of the disease. However, the consistent use of fungicides has resulted in the development of resistance in many red pepper-growing areas of Korea. Effective management of the occurrence of fungicide resistance depends on constant monitoring and early detection. Thus, in this study, various methods such as agar dilution method (ADM), gene sequencing, allele-specific polymerase chain reaction (PCR), and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were applied for the detection of benzimidazole resistance among 24 isolates of Colletotrichum acutatum s. lat. and Colletotrichum gloeosporioides s. lat. The result of the ADM showed that C. gloeosporioides s. lat. was classified into sensitive and resistant isolates to benomyl while C. acutatum s. lat. was insensitive at ≥1 ㎍/ml of benomyl. The sequence analysis of the β-tubulin gene showed the presence of a single nucleotide mutation at the 198th amino acid position of five isolates (16CACY14, 16CAYY19, 15HN5, 15KJ1, and 16CAYY7) of C. gloeosporioides s. lat. Allele-specific PCR and PCR-RFLP were used to detect point mutation at 198th amino acid position and this was done within a day unlike ADM which usually takes more than one week and thus saving time and resources that are essential in the fungicide resistance management in the field. Therefore, the molecular techniques established in this study can warrant early detection of benzimidazole fungicide resistance for the adoption of management strategies that can prevent yield losses among farmers.

• 한국어병학회지
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• 제23권1호
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• pp.85-98
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• 2010

농림수산식품부와 국립수산과학원에서는 2009년도 방류 품종 (해면 품종 22종과 내수면 품종 11품종)을 대상으로 수산동물전염병의 감염 여부를 검사하였다. 총 12개 지방자치단체에 서 방류를 실시한 것으로 나타났으며, 이 중에서 경상남도, 전라남도, 제주도 및 충청남도는 해면 품종을 주로 방류하였으며, 경기도, 전라북도 및 충청북도는 내수면 품종을 많이 방류하는 것으로 나타났다. 검사품종중에서 해산품종으로는 전복이 24.5%로 가장 많았으며, 그 다음 해삼(15.2%), 넙치 (11.5%), 감성돔과 조피볼락(6.8%), 꽃게 (5.6%), 돌돔 (5.1%), 볼락 (4.6%), 붉은쏨뱅이 (4.5%)로 나타났다. 내수면 품종 중에서는 붕어가 19.4%로 가장 많았으며, 그 다음으로 뱀장어 (17.0%), 동자개 (12.3%), 다슬기 (12.0%), 메기 (8.4)의 순으로 검사 실적이 많았다. 총 33종의 품종을 대상으로 1,080회의 검사가 의뢰되었으며, 검사항목별로 2,066건의 검사를 실시한 결과 19건에서 red sea bream iridovirus (RSIV), koi herpesvirus (KHV) 또는 white spot syndrome virus (WSSV)와 같은 병원체가 검출되어 불합격 처리되었다.