• The Journal of Korean Society of Virology
• /
• v.30 no.1
• /
• pp.29-37
• /
• 2000

The aim of this study was to investigate viral etiology in dilated cardiomyopathy (DCM) by polymerase chain reaction (PCR) or nested reverse transcription PCR (RT-PCR), and characterize the enteroviral RNA presented in the clinical specimens. Twenty-eight paraffin-embedded heart tissue samples were assayed to detect cytomegalovirus, herpes simplex virus type 1, type 2, parvovirus, adenovirus, and enterovirus (EV) with each specific primer. Of these 28 patients (mean age: 27, M: 24, F: 4), 26 were histologically diagnosed as DCM and 2 as myocardial infarction (MI). Nested RT-PCR detected enteroviral RNA in 7 (26.9%) of 26 patients with DCM, and none of patients with MI. And none of DNA viruses tested were detected from the samples. Amplified products were also genotyped by single-strand conformation polymorphism (SSCP). Three subtypes can be differentiated from 7 clinical specimens. Furthermore, direct sequence analysis was performed to determine whether genetic variation of EV is present in the explanted heart tissues from patients with DCM. Although most of the sequences among the wild isolates have the greatest similarity to those of coxsackievirus B3, there are specific regions of variable sequences (no 490 - no 510). The data suggest that enterovirus may be a major viral pathogen for the DCM in Korea and nucleotide sequence data indicate that coxsackievirus B3 may be a leading etiologic agent of DCM.

• Parasites, Hosts and Diseases
• /
• v.53 no.3
• /
• pp.315-320
• /
• 2015

Acarapis mites, including Acarapis woodi, Acarapis externus, and Acarapis dorsalis, are parasites of bees which can cause severe damage to the bee industry by destroying colonies and decreasing honey production. All 3 species are prevalent throughout many countries including UK, USA, Iran, Turkey, China, and Japan. Based on previous reports of Acarapis mites occurring in northeast Asia, including China and Japan, we investigated a survey of Acarapis mite infestations in honey bees in Korean apiaries. A total of 99 colonies of Apis mellifera were sampled from 5 provinces. The head and thorax of 20 bees from each colony were removed for DNA extraction. PCR assays were performed with 3 primer sets, including T, A, and K primers. Results indicated that 42.4% (42/99) of samples were Acarapis-positive by PCR assay which were sequenced to identify species. Each sequence showed 92.6-99.3% homology with reference sequences. Based on the homology, the number of colonies infected with A. dorsalis was 32 which showed the highest infection rate among the 3 species, while the number of colonies infected with A. externus and A. woodi was 9 and 1, respectively. However, none of the Acarapis mites were morphologically detected. This result could be explained that all apiaries in the survey used acaricides against bee mites such as Varroa destructor and Tropilaelaps clareae which also affect against Acarapis mites. Based on this study, it is highly probable that Acarapis mites as well as Varroa and Tropilaelaps could be prevalent in Korean apiaries.

• Korean Journal of Veterinary Research
• /
• v.47 no.1
• /
• pp.33-41
• /
• 2007

Streptococcus spp. comprising Streptococcus (S.) uberis S. dysgalactiae strains is major causeof bovine mastitis from particularly well-managed or low somatic cell count herds that have successfullycontrolled contagious pathogens. In this study, antimicrobial susceptibility and genetic characteristics of S.uberis isolated from clinical or subclinical mastitis milk at 2003 were investigated. Eighty seven isolatesof Streptococus spp. were identified by the conventional biochemical methods. The antimicrobialsusceptibility by disk diffusion method was determined for 46 S. uberis, 11 S. bovis, 10 S. oralis, 6 S.uberis and 14 other Streptococcus spp.. Overall, the tested strains were susceptible to tetracycline (11.5%),amikacin (14.9%), streptomycin (16.1%), neomycin (26.4%), kanamycin (35.6%), gentamicin (65.2%),oxacillin (70.1%), ampicillin (75.9%), chloramphenicol (78.2%), and cephalothin (97.7%). Additionally, S.uberis strains were susceptible to pencillin G (97.8%), but resistant to erythromycin (76.0%) by minimalinhibitory concentration test. The multiple-drug resistance rate of isolated bacteria to 4 more thanamplification fingerprinting patterns amplifed with primer 8.6d showed that 3 to 8 number of distinguishableDNA fragments ranged from 180 bp to 1,20 bp. Thirty seven isolates of S. uberis strains were subtypedinto 8 distinct patterns. Each subtype revealed a typical pattern of antimicrobial susceptibilities. Thesefindings demonstrate that S. uberis isolates were mastitis pathogens of diverse serotypes, and oftenencountered the diverse resistant patterns.

• Research in Plant Disease
• /
• v.15 no.2
• /
• pp.88-93
• /
• 2009

In 2008, leaf rot and blight on pepper seeding ("Dokya-chungchung") occurred in a pepper farm at Hwaseong-si, Gyeonggi-do, Korea. The typical symptom is water-soaking and dark brown leaf blight at edges and tips of leaves. The fungal colonies isolated from infected tissues were pinkish at first and turned gradually to gray. Conidia were fusiform, non-septum, and $8.1-17.0{\times}2.0-3.8{\mu}m$ in size. Several specific PCR primers derived from the sequence of the internal transcribed spacer (ITS) region of the rDNA, such as CaINT, CgINT and CcINT were used for the identification of the fungal pathogen. The C. acutatum-specific primer CaINT was amplified single fragment of 496 bp that discriminated C. acutatum from the other species. The pathogenicity test was performed on seedlings and fruits of red pepper. On the basis of the morphological, molecular characteristics and pathogenicity test, we identified as Colletotrichum acutatum. This is the first report on leaf rot and blight on pepper seedling caused by C. acutatum in Korea.

• Journal of Microbiology and Biotechnology
• /
• v.15 no.3
• /
• pp.502-509
• /
• 2005

Sexual reproduction plays an important role in the biology and epidemiology of oomycete plant pathogens such as the heterothallic species Phytophthora infestans. Recent worldwide dispersal of A2 mating type strains of P. infestans resulted in increased virulence, gene transfer, and genetic variation, creating new challenges for disease management. To develop a genetic assay for mating type identification in P. infestans, we used the Amplified Fragment Length Polymorphism (AFLP) technique. The primer combination E+AT/M+CTA detected a fragment specific to A1 mating type (Mat-A1) of P. infestans. This fragment was cloned and sequenced, and a pair of primers (INF-1, INF-2) were designed and used to differentiate P. infestans Mat-A1 from Mat-A2 strains. The Mat A1-specific fragment was detected using Southern blot analysis of PCR products amplified with primers INF-1 and INF-2 from genomic DNA of 14 P. infestans Mat-A1 strains, but not 13 P. infestans Mat-A2 strains or 8 other isolates representing several Phytophthora spp. Southern blot analysis of genomic DNAs of P. infestans isolates revealed a 1.6 kb restriction enzyme (EcoRI, BamHI, AvaI)-fragment only in Mat-A1 strains. The A1 mating type-specific primers amplified a unique band under stringent annealing temperatures of $63^{\circ}C-64^{\circ}C$, suggesting that this PCR assay could be developed into a useful method for mating type determination of P. infestans in field material.

• Journal of Life Science
• /
• v.8 no.2
• /
• pp.126-130
• /
• 1998

A pair of designed primers (sequences from Gene Bank) amplified 16S fRNA gene of V. vulnificus within polymerase chain reaction (PCR) machine. This PCR product is about 1.3kb DNA fragment. Six enzymes (BamH I, Alu I, Sau3A I, Hind III, Sal I, Sma I) were used for restriction pattern analysis of amplified 16S rRNA gene of V. vulnificus ATCC 27562. Digested fragments are resolved by 3% agarose gel. BamH I did not show digested fragment so, there was no cutting site of BamH I in PCR product. Alu I produced three small fragments from 400 bp to 200 bp. Sau3A I produced three fragments larger than Alu I from 70 bp and 500 bp. One of fragments of Sal I was same with 500 bp of Hind III fragment and the other was 750 bp. Sma I showed two fragments of 800 bp and 470 bp. The profile of digested fragments of 16S rRNA of V.vulnificus ATCC 27562 will may be able to use standard profile for identification of V. vulnificus.

• Clinical and Experimental Reproductive Medicine
• /
• v.23 no.3
• /
• pp.341-349
• /
• 1996

초기배의 성판정은 대상가축의 성을 선발하는 수단으로써 뿐만아니라 인간의 유전적 질병의 조기진단법으로서 매우 가치가 크다. 체외수정 소 초기배의 성을 결정하기 위해 PCR과 PRINS를 이용하였으며 성판정에 이용된 8 세포${\sim)$배반포기 초기배는 체외수정후 난관상피세포와의 공배양에 의해 생산되었다. 초기배 의 DNA는 $200{\mu}g/ml$ proteinase K가 함유된 PCR lysis buffer에 하나의 초기배를 부유한 후 $50^{\circ}C$에서 1시간동안 처리하여 준비하였다. 중기 염색체 spreads는 초기배를 nocodazole로 처리한 후 air-drying 방법을 이용하여 준비하였다. 가능한 false positive signals을 배제하기 위해 소특이 및 Y 염색체 특이 primers를 이용하여 PCR을 수행한 결과, 웅성 초기배에서는 두 개의 증폭산물 (소특이 및 Y 염색체 특이)이 합성된 반면 자성 초기배에서는 하나의 증폭산물만 합성되었다. 한편 중기염색체상의 Y 염색체를 동정하기 위해 FISH와 PRINS를 수행한 결과, FISH에서보다 PRINS에서 더 강한 Y 염색체 특이 형광 signals이 탐지되었다. 이러한 결과는 PCR에 의한 체외생산 소 초기배의 신속정확하고 효율적인 성판정이 가능함을 보여주었다. 또한 PRINS를 통해 PCR 에 이용된 Y 특이 probe의 신뢰성이 염색체 수준에서 확인되었다.

• 한국균학회소식:학술대회논문집
• /
• 2014.10a
• /
• pp.41-41
• /
• 2014

Agaricus bisporus, commonly known as the button mushroom, is the most widely cultivated species of edible fungi. Low frequency of recombination ratio and homokaryotic or monokaryotic spore on meiotic basidia form obstacles for breeding programs. Since the first hybrid varieties for white button mushrooms were released in Europe, new varieties released afterwards were either identical of very similar to these first hybrids on morphologies. Therefore, different DNA markers have been used to define unique varieties of A. bisporus strains. Aim of this study is to assess the genetic diversity of different A. bisporus strains in Korea. Twelve UFP (Universal fungal primer, JK BioTech. Ltd), 12 simple sequence repeat (ISSR) and 30 SSR primers were used to assess genetic diversity of monokaryotic and dikaryotic Agaricus bisporus strains including other 19 Agaricus spp. Of them, four UFP, four SSR primers, $(GA)_8T$, $(AG)_8YC$, $(GA)_8C$ and $(CTC)_6$ and seven SSR markers produced PCR polymorphic bands between the Agaricus species or within A. bisporus strains. PCR polymorphic bands were inputted for UPGMA cluster analysis. Forty five strains of A. bisporus are genetically clustered into 6 groups, showing coefficient similarity from 0.75 to 0.9 among them. In addition, genetic variations of monokaryotic and dikaryotic Agaricus bisporus strains were partially detected by PCR technologies of this study. The varieties, Saea, saedo, Saejeong and Saeyeon that have recently been developed in Korea were involved in the same group with closely genetic relationship of coefficient similarity over 0.96, whereas, other strains were genetically related to A. bisporus strains that were introduced from USA, Eroupe and Chinese.

• Korean Journal of Veterinary Research
• /
• v.38 no.1
• /
• pp.173-181
• /
• 1998

Esherichia coli O157 : H7의 slt I, slt II, uid A 및 eaeA 4종 유전자를 동시에 검출하기 위한 multiplex PCR 기법을 확립하고 쇠고기중 직접 E coli O157 : H7 검출시험을 실시하였다. 4 set의 primers를 이용한 multiplex PCR 기법으로 31종의 장내세균에 대한 특이성을 조사한 결과 E coli O157 : H7 에서 1,087bp (eae A), 584bp (slt II), 348bp (slt I) 또는 252bp (uid A)크기의 DNA를 동시에 특이적으로 검출할 수 있었다. E coli O157 : H7 15주는 모두 uid A 및 eae A 유전자가 동시에 검출되었고, 다른 장내세균에서는 검출되지 않았다. slt I 또는 slt II 유전자를 가지고 있는 E coli 표준균주 24종을 이용하여 multiplex PCR 기법과 Vero cell cytotoxicity assay을 비교검사한 결과 베로톡신 산생능과 PCR법의 결과는 일치하였다. mutiplex PCR 기법의 쇠고기중 검출한계는 modified EC(mEC)에서 증균없이는 E coli O157 : H7균 $10^4cells/g$ 이상에서 검출이 가능하였으나 mEC에 1차 증균후 modified TSB 증균하였을 경우에는 10cells/g이하까지도 검출이 가능하였다. 개발된 multiplex PCR 기법을 쇠고기 40종에 직접 적용한 결과 E coli O157 : H7은 검출되지 않았으나 slt I 및 slt II유전자를 가지고 있는 E coli 4종이 검출되었으며, 이들의 혈청형은 O6, O112, O115 및 O139 였다. 이 연구에서 개발된 multiplex PCR은 쇠고기중 E coli O157 : H7을 신속하고 특이적으로 검출하는데 사용할 수 있을 것으로 사료된다.

• Korean Journal of Plant Resources
• /
• v.22 no.5
• /
• pp.446-453
• /
• 2009

RAPD analyses were performed to investigate genetic relationships and useful molecular maker for 3 species and their 17 regional populations of the Pueraria lobata and related taxa. The length of amplified DNA fragments ranged from 200 to, 2,800 bp. Two hundred and eight scorable polymorphic makers and three scorable monomorphic makers were found from the PCR reactions with 15 random oligo primers, and those were analyzed by Nei's genetic distance coefficient. Based on the UPGMA phenogram from RAPD analyses, two major groups (9 populations from Korea; 3 populations from foreign countries) were recognized. And it showed distinct genetic differences from related taxa. The RAPD results was very useful to define the samples by geographical distribution and to discuss the relationships among the populations and their related taxa of the Pueraria lobata.