미세방울 디지털 PCR(Droplet digital PCR, ddPCR) 법은 정량 PCR 법과 같이 형광물질을 사용하여 정량분석이 가능하다는 점은 유사하지만, PCR 반응액을 만든 후 이 반응액을 바로 PCR에 사용하지 않고 수 만 개의 동일한 크기의 미세방울(droplet)로 구획(partitioning)한 후에 PCR 반응을 수행하는 점이 기존의 PCR 법과 가장 큰 차이라고 볼 수 있다. 이렇게 구획된 나노리터 크기의 미세방울 중에서 형광신호가 검출되는 미세방울과 검출되지 않은 미세방울의 수를 계수하여 프아송 분포(Poisson distribution) 계산에 적용하면 표준검량선 없이 목적 유전자의 절대정량 분석이 가능하다. 곤충이 매개하는 질병 바이러스의 경우 소량의 바이러스 감염 여부를 확인하기 위해서 다수의 곤충 유전자를 확인해야 하는데, 이와 같이 미세방울 형태의 ddPCR을 이용하면 기존 Real-time PCR 법에 비해 극소량의 목적 유전자를 높은 민감도로 검출할 수 있으며, PCR 저해요소(inhibitor)에도 큰 영향을 받지 않는다는 장점이 있다. 또한 미세방울 방식의 디지털 PCR을 이용하면 다중 PCR 분석이 가능하여 1개의 시료에서 다양한 질병매개 바이러스를 검출할 수 있다.

Korean horn beetles, Allomyrina dichotoma (Linnaeus, 1771) reared in many local farms are suffering from a fatal viral disease as reported in 2015, and recent sequencing analysis revealed that the virus is very closely related to Oryctes rhinoceros nudivirus. In a nationwide investigation, it was concluded that the virus can transmit vertically from an infected adult to the offspring by indirect manner, and 70 to 77% of young larvae dead in early stage were diagnosed as this virus positive. Here, we report for the establishment of on-site diagnosis method for the viral disease using loop-mediated isothermal amplification (LAMP) assay. A. dichotoma larvae were assayed for the OrNV infectivity with LAMP primers targeting OrNV_gp102 gene. To evaluate the LAMP specificity, two bacterial pathogens, Bacillus thuringiensis and Serratia marcescens, causing disease in A. dichotoma were tested along with OrNV. Only from the OrNV-infected larvae the reaction resulted positive. Also, to avoid DNA extraction process, the LAMP assay used diluted hemolymph directly and 50-fold dilution was set for diagnosis standardization and convenient on-site application for infected larvae from local farms. The LAMP diagnosis is fast and convenient for nontechnical user in the field and expected to stop this foreign virus spread all over the country.

구제역은 발생 시 전염이 쉽게 일어나며 심각한 경제적 피해를 일으키는 질병이다. 구제역의 방역정책은 발견 직 후 빠른 살처분이 최선책이나, 전파 속도나 상황 등에 따 라 타지역 백신 접종 등의 방법을 시행할 수도 있다. 이러 한 방법을 적용하기 위해서는 구제역을 빠르고 정확하게 진단할 필요성이 있다. 개발된 진단법들은 구제역의 확진, 혈청형의 동정, 백신 접종 후 항체의 생성 확인 등에 사용 된다. 많은 진단법들이 개발되었지만 아직은 빠른 시간 내 에 검출이 가능하며 동시에 정확성도 가진 방법이 드물다. 그렇기에 기존의 방법들을 개선시킨 새로운 진단법이 필 요하다. 현재는 대부분 혈청학적 진단법인 ELISA에 의존 하거나 분자 유전학적 기술인 PCR을 사용한다. 가장 최근 기술은 그 둘을 합치는 방법으로, 어떻게 하면 더 신속하 고 저비용이면서, 민감하고 정확한 방법이 될 수 있을지 연구가 진행되고 있다.

Virus infections of the honeybee(Apis mellifera) have been increasingly investigated during the last decade. In general, honeybee viruses are widespread and most of them persist as inapparent infections. We screened honeybee colonies for the presence of several bee viruses, including deformed wing virus(DWV), black queen virus(BQCV), Kashmir bee virus(KBV), Israeli acute paralysis virus (IAPV), sacbrood virus(SBV), acute bee paralysis virus(ABPV), using uniplex RT-PCR. Frequently simultaneous infections with different viruses are diagnosed in seemingly healthy bee colonies. Therefore we developed a multiplex RT-PCR assay for the simultaneous detection of multiple bee viruses.

Human voice reacts very sensitively to human's minute physical condition For instance, human voice disorders affect patients, profoundly especially in the case of Parkinson's disease Acoustic tools such as MDVP, can function as an equipment that measures

In this chapter 8, I discuss a running piglet disease(奔豚氣病), its causes, symptoms and prescription. The name of the running piglet(奔豚) disease dates back to old times, and it is not clinically reported these days. The symptoms of the running piglet disease(奔豚氣病) is characterized by unberable pain in the lower abdomen, the chest and the throat, followed by complete disappearance of these symptoms with passage of time. In my view, the causes of the running piglet disease(奔豚氣病) can be divided into three categories: startle and panic, and the excessive fluid(水飮). In this chapter 13, I elaborate on three diseases: a wasting-thirst(消渴), a dribbling urination(小便不利),1)a strangury disease(淋病). I discuss these three diseases in one chapter because they share the same symptoms of thirst and abnormal urination and the symptoms occur mainly in the kidney(腎) and the urinary bladder(膀胱). A wasting-thirst(消渴) has the main symptoms of drinking water and eating food excessively as well as urinating a lot. A disease symptomized by thirst(消渴) is typically divided into upper wasting-thirst(上消), middle wasting-thirst(中消), and lower wasting-thirst(下消) by clinicians. Specifically, I discuss the mechanism of disease of upper wasting-thirst(上消) and lower wasting-thirst(下消), treatment principles of and prescriptions for upper wasting-thirst(上消) and lower wasting-thirst(下消), and pathology of middle wasting-thirst(中消) in this chapter. Based on the view that a dribbling urination(小便不利) is not a disease per se but a label for the symptom of short and insufficient urination, I discuss water amassment syndrome(蓄水證), yang bringtness disease pattern(陽明病), and a dribbling urination(小便不利) that is accompanied by various diseases. A strangury disease(淋病) has the main symptoms of insufficient and trickling urination that is often accompanied by pain perception. In this chapter, I discuss about stone strangury(石淋) in particular. These three diseases merit in-depth discussion since they are clinically observed with high frequency.

Background : Development of new real-time PCR diagnosis method for simultaneous diagnosis of Cylindrocarpon destructans and Fusarium solani, causative fungi of ginseng root rot disease. C. destructans and F. solani are known to be the major pathogens of ginseng root rot disease. Root rot caused by these pathogens is a disease that is difficult to control because the disease progresses slowly and it is difficult to diagnose early and even when symptoms of plant seeding are present, the disease is already spread in the roots. Diagnostic methods to detect the presence or absence of ginseng roots rot fungi in soil before ginseng cultivation are currently being used as a method for controlling. However, commercialized soil extraction kits and PCR diagnostics have cost, diagnostic time, and single diagnostic problems, and need to develop new diagnostic methods. Methods and Results : Primers and probes in the beta-tubulin 2 gene were designed for species-specific detection. In silico analysis, the detection rate of C. destructans was 100% and the detection rate of F. solani was 95%. The multiplex real time PCR optimization conditions including the internal control were established. The analytical sensitivity using positive samples was 10 copies/㎕ for C. destructans and 10 copies/㎕ for F. solani. As a result of performance comparison test with conventional PCR diagnosis methods, it was confirmed that the developed multiplex real time PCR method has the same or better performance in terms of sensitivity. In the developed soil extraction kit, the extraction time was reduced and the extracted DNA quality was improved, compared to the used soil extraction kit. Conclusion : From the above results, we expect that the developed C. destructans / F. solani multiplex real time PCR diagnosis method and soil extraction kit will be useful for real-time monitoring of ginseng root rot pathogenic fungi in the soil of ginseng cultivation area and diagnosis of suitability of ginseng cultivation area.

Chronic kidney disease can be treated if it is detected early, but as the disease progresses, it becomes impossible to recover. Finally, renal replacement therapy such as transplantation or dialysis should be used. Ultrasonography is used to diagnose kidney cancer, inflammatory disease, nodular disease, and chronic kidney disease. It is used to identify information about degree of inflammation using information such as kidney size, internal echo characteristics. Currently, the degree of disease in the clinic uses the value of glomerular filtration rate. However, even in ultrasound, changes in the degree of inflammation and disease can be observed. In this study, we used ultrasound images to quantify the changes in brightness, size, cortex, and subclinical changes of the kidney with progression of the disease, and compared them with the glomerular filtration rate used in clinical practice . In 105 cases, we performed 35 cases of normal kidney, 35 cases of early kidney disease, and 35 cases of terminal kidney. The brightness of the cortex of the image was obtained and the difference in brightness between the cortex and the proximal portion was obtained by the slope. The graph of the portion which was not smooth due to the ultrasonic characteristics was used as the function regrass. The size reduction was obtained from the original data. The results were as follows: It was proportional to the glomerular filtration rate. It is considered that the algorithm can be applied to the disease if the algorithm study continues.

Rosai-Dorfman Disease is a very rare systemic immunologic disease, but most patients show improvement without specific treatment. However, when confined to the breast, it is difficult to exclude from malignancy since they are share similar radiographic characteristics. Thus, we report a case of Rosai-Dorfman Disease confined to the breast that pathological confirmation is needed to diagnose the disease via core needle biopsy or surgical excision.

알츠하이머병은 치매를 일으키는 가장 흔한 퇴행성 뇌 질환이다. 뇌에 축적되는 베타 아밀로이드 단백질은 기억력감퇴, 언어능력 저하등 일상생활 수행 능력이 어렵게 된다. 베타 아밀로이드 플라그 농도는 인지 장애를 가진 성인환자에서 알츠하이머 질환과 인지장애의 다른 원인인지를 평가하는 데 사용된다. 베타아 밀로이드 단백질에 대한 높은 민감성과 특이성을 가진 18F-Florbetaben을 이용하여 알츠하이머병을 조기진단에 유용성을 알아보고자 한다. 18F-FDG Brain 영상에서 특이소견 없음을 보인다. 그리고 MR-Brain 영상에서 해마의 위축이 없는 것으로 보였다. 하지만 18F-Florobetaben에서 베타 아밀로이드의 섭취는 알츠하이머 병의 진전이 되고 있음을 알려준다. 따라서, 18F-Florobetaben은 알츠하이머병을 조기 진단하는 데 매우 유용하다.

본 연구는 간 초음파영상에서 통계적 속성 기반의 밝기 히스토그램에 기초한 픽셀 질감분석 파라미터(평균밝기, 왜곡도, 균일도, 엔트로피)와 간과 콩팥실질의 밝기 차를 이용한 영상분석을 통해 미만성 간질환의 컴퓨터보조진단 적용 가능성을 알아보고자 하였다. 실험은 간 초음파영상(정상, 지방간, 간경화)에서 관심영역(50×50픽셀)을 설정하고 4가지의 픽셀 질감분석 파라미터와 간과 콩팥의 실질 밝기의 차를 이용하여 질환인식률을 평가하였다. 그 결과 평균밝기, 균일도, 엔트로피의 질환인식률은 100%, 왜곡도 96%로 높게 나타났으며, 간과 콩팥의 실질 밝기 차는 정상 -1.129±12.410, 지방간 33.182±11.826으로 뚜렷한 차이를 나타내었으나, 간경화의 경우 -1.668±10.081로 정상과는 다소 작은 차이를 나타내었다. 이러한 결과를 바탕으로 높은 질환인식률을 보인 픽셀 질감분석 파라미터와 실질 밝기 차를 이용한 컴퓨터보조진단은 미만성 간질환의 감별에 유용한 도구로써 임상적인 활용 가능성이 있으며, 판독 오류를 최소화하고 정확한 진단과 치료방향 제시에 도움이 될 것으로 기대된다.