Salmonella spp.는 식중독의 주요 원인균으로, 신속하고 정확한 검출 방법이 요구된다. 본 연구에서는 간소화된 direct multiplex real-time PCR 방법인 FS Finder SL키트 의 분석법 활용 가능성을 평가하고, 평판배지법과의 검출 성능을 비교하였다. 또한, real-time PCR의 검출 효율을 평 가하기 위해 FS Finder SL 키트에서 제공하는 세 가지 전 처리 방법(Method 1, 2, 3)을 비교 분석하였다. 실험 결과, 세 가지 전처리 방법을 이용한 direct multiplex real-time PCR 방법은 Salmonella spp.를 100% 검출할 수 있었으며, Ct 값 비교를 통한 통계적 분석에서도 세 방법 간 유의한 차이가 없는 것으로 나타났다(P>0.05). 반면, 선택 배지를 이용한 검출에서는 2 log CFU/g 이상으로 접종된 샘플에 서만 Salmonella spp.가 검출되었으나 real-time PCR법의 경우 0-3 log CFU/g 범위의 샘플에서 모두 검출이 가능 하였다. 또한, 실험에 사용된 세 가지 즉석섭취식품군(알 가공품, 닭가슴살 제품, 편의점 도시락)에서 자연균총의 영 향을 평가한 결과, 도시락 샘플의 일반세균수가 3.56±0.18 log CFU/g으로 가장 높았으며, 알가공품과 닭가슴살 제품 에서는 검출되지 않았다. 결론적으로 FS Finder SL 키트 를 활용한 real-time PCR 방법은 기존의 평판배지법보다 높은 검출 감도를 보였으며, 검출까지의 소요 시간을 대 폭 단축할 수 있었다. 특히, 복합적인 식품 매트릭스에서 도 신속하고 정밀한 검출이 가능함을 확인하였다. 본 연 구는 즉석섭취식품 중 Salmonella spp. 검출을 위한 효율 적인 direct multiplex real-time PCR 분석법의 적용 가능 성을 제시하며, 향후 식품안전 관리 시스템에서 활용될 수 있는 기초 자료를 제공한다.

Monochamus alternatus (M. alternatus) and Monochamus saltuarius (M. saltuarius) are major vectors for Bursaphelenchus xylophilus in South Korea. When an adult, they are easily distinguishable by several morphological classification. However, it is difficult to identification between M. alternatus and M. saltuarius when they are larvae as they have very similar morphological characters. Thus, they are not easily distinguishable without expertise about Cerambycidae taxonomy. Furthermore, during epidemiological investigation, sometimes, adults or larvae would not be founded in death pine trees. For these reasons, in this experiment, we are able to identified between M. alternatus and M. saltuarius by mitochondrial 12S rRNA gene primers that are specific to 12S rRNA gene fragment of M. alternatus using larvae tissue and frass. Moreover, we had examined whether vectors that were already escaped from dead pine tree have Bursaphelenchus xylophilus or not by multiplex PCR using larva frass that was remained in dead pine tree.

It is difficult to identification between Bursaphelenchus spp. and Pine Wood Nematode (PWN) by morphological characteristics without expertise about nematode taxonomy. Furthermore, Baermann funnel method, which is nematode extraction method from wood chips or soil, requires at least 24 hours to extract nematode that is unsuitable to rapid diagnose the Pine Wilt Disease (PWD). For these reasons, the aim of this experiment is not only to improve accuracy of a PCR based method but also to reduce total experiment time for detection Bursaphelenchus spp. and PWN in the wood chips of PWD infected pine tree. In this experiment, we had been employed two PCR primer sets, which were originated from PWN specific Internal Transcribed Spacer (ITS) sequence region and Bursaphenchus spp. universal mitochondrial Cytocrome Oxidase subunit I (mtCOI) sequence region in order to discrimination between Bursaphelenchus spp. and PWN at the same PCR reaction. This experimental procedure was able to reduce experiment time and cost as well as to improve accuracy of detection than previous PCR based detecting method by not using Baermann funnel method and commercial genomic DNA extraction kit but using direct pine wood chips lysis method.

Lepidopteran pests monitoring in adult stage was generally performed using delta or corn typed trap including rubber septa impregnated sex pheromone (lure). Sometimes, unfortunately trapped samples were severly damaged because of biotic and/or abiotic environments such as micro-organism, predator and rain, sticky material, respectively. In our case, we monitored potato tuber moth, PTM, Phthorimaea operculella distribution during 2009~2012 in Korea. However, we encountered unexpected problem, another species can be trapped in species specific sex pheromone trap. Therefore, species confirmation was needed in trapped samples. Here we developed confirmation method by direct PCR (without DNA extraction) or sequencing methods which trapped samples that cannot identified by morphologically. We designed multi-plex PCR universal primers and species specific primers in rRNA region because to check the success of PCR and species identification. This direct PCR method can be applied in other species confirmation which monitored using pheromone trap.

The mating-type genes control formation of the dikaryon from two haploid strains. These genes are now used in mating-type-assisted breeding programs for economically important mushrooms, especially the oyster mushroom, Pleurotus ostreatus, aiming at high-yield and high-quality standard mushroom production. However, it improves the breeding program when the breeder is able to quickly identify compatible strains in a given set of progeny. The two mating factors with their mating-type loci are used as markers for breeding and have been incorporated in a chromosome mapping investigation. The linkage maps include not only genetic markers such as the mating types that can be cored, but also molecular markers such as PCR-assisted approaches, e.g. RAPD analyses, or RFLP markers. Once mating-type genes within progeny may be more easily identified by the use of PCR-directed cloning of partial mating-type genes. We analyzed homeodomain (HD1 and HD2) and pheromone receptor(rcb1, 2 and 3) genes as molecular markers for breeding using mating type A and B of Pleurotus eryngii and Pleurotus ferulae by direct PCR.