In this critical review the applications of gold catalysed reactions in total synthesis during the years since our last article are reviewed. At the end of this article a literature analysis is conducted to evaluate the progress in this field.

Gold catalysis in total synthesis – recent achievements

Loop-mediated isothermal amplification (LAMP) has become a powerful alternative to polymerase chain reaction (PCR) for pathogen detection in clinical specimens and food matrices. Nontyphoidal Salmonella is a zoonotic pathogen of significant food and feed safety concern worldwide. The first study employing LAMP for the rapid detection of Salmonella was reported in 2005, 5 years after the invention of the LAMP technology in Japan. This review provides an overview of international efforts in the past decade on the development and application of Salmonella LAMP assays in a wide array of food and feed matrices. Recent progress in assay design, platform development, commercial application, and method validation is reviewed. Future perspectives toward more practical and wider applications of Salmonella LAMP assays in food and feed testing are discussed.

/pdf/loop-mediated-isothermal-amplification-for-salmonella-v5c7ygyj1m.pdf

Loop-Mediated Isothermal Amplification for Salmonella Detection in Food and Feed: Current Applications and Future Directions.

Formal Total Synthesis of Spirangien A

We developed a loop-mediated isothermal amplification (LAMP) method to rapidly diagnose Wheat streak mosaic virus (WSMV) during quarantine inspections of imported wheat, corn, oats, and millet. The LAMP method was developed as a plant quarantine inspection method for the first time, and its simplicity, quickness, specificity and sensitivity were verified compared to current reverse transcription-polymerase chain reaction (RT-PCR) and nested PCR quarantine methods. We were able to quickly screen for WSMV at quarantine sites with many test samples; thus, this method is expected to contribute to plant quarantine inspections.

Loop-mediated Isothermal Amplification Assay to Rapidly Detect Wheat Streak Mosaic Virus in Quarantined Plants

Molecular Genetics and Subcellular Localization of Flavonoid Metabolism in Arabidopsis

살모넬라는 음식과 식수에서 흔히 나오는 중요한 병원체로 세계 곳곳에서 급성 위장염과 같은 감염증을 일으키며, 일반적으로 인간의 혈청형 임상종으로는 Salmonella enterica의 혈청형인 S. Typhimurium과 S. Enteritidis가 있다. 일반적인 검출 방법으로 살모넬라를 기본으로 하여 선택적인 배양으로 샘플을 수집하였고 살모넬라를 일으키는 군체의 특징을 생화학과 혈청학상인 테스트를 하였으나 이러한 방법들은 일반적으로 시간이 걸리고 높은 감도를 보이지 않았다. 최근, 등온증폭반응법과 real-time PCR법을 이용하여 높은 감도, 특이성으로 현재 병원성 박테리아에 빠르게 수행할 수 있게 되었다. 본 연구에서는 등온증폭반응과 real-time PCR법을 사용하여 S. Typhimurium과 S. Enteritidis의 검출하였다. 선택적인 타겟 유전자로, invA를 살모넬라종의 염기서열에 특이적으로 임의복제 하였다. 등온증폭반응과 real-time PCR은 살모넬라종으로부터 임의의 염기서열을 증폭하여 검출하였고, invA는 S. Typhimurium과 S. Enteritidis의 두 가지 종을 모두 검출하였다. 이러한 등온증폭반응과 real-time PCR법으로 S.Typhimurium과 S. Enteritidis의 검출 가능성을 보였으며, 살모넬라 종에 대한 특이성, 민감성을 갖춘 유용한 검출방법을 제시하였다. 【Salmonella is an important food-and water-borne pathogen associated with acute gastrointestinal illnesses around the world. The most common serotypes isolated from humans are Salmonella enterica serotype Typhimurium (S. Typhimurium) and S. Enteritidis. Traditional detection methods for Salmonella are based on cultures using selective media and characterization of suspicious colonies by biochemical and serological tests. These methods are generally time-consuming and not so highly sensitive. Recently, the Loop Mediated Isothermal Amplification and real-time PCR has been used as a highly sensitive, specific, and rapid test for the presence of pathogenic bacteria. In this study, a LAMP and real-time PCR was used to detect S. Typhimurium and S. Enteritidis. We selected target genes, which were the in invA and a randomly cloned sequence specific for the genus Salmonella. With LAMP and real-time PCR, random sequence was detected from Salmonella spp, invA were detected from all strain of S. Typhimurium and S. Enteritidis. This assay indicate that the specificity, sensitivity and rapid of the LAMP and real-time PCR make them potentially valuable tools for detection of S. Typhimurium and S. Enteritidis.】

/pdf/detection-of-salmonella-using-the-loop-mediated-isothermal-16gltki722.pdf

Detection of Salmonella Using the Loop Mediated Isothermal Amplification and Real-time PCR

. In the meat industry, correct breed information in food labeling is required to assure meat quality. Genetic markers provide corroborating evidence to identify breed. We described the development of DNA markers to discriminate between Korean beef cattle (Hanwoo), Holstein, and mixed cow beefs. As most breeds are standardized for coat colour, the melanocortin 1 receptor (MC1R) gene, involved in the regulation of eu/pheomelanins synthesis, has been suggested as marker for breed traceability of products of animal origin. We also designed sex-determining region Y (SRY) gene specific primers for Y chromosome detection. In thisstudy, fragments of MC1R gene and SRY gene were amplified by multiplex-PCR and subjected to digestion by Msp A1I restriction endonuclease. Reaction products were analysised by denaturing high performance liquid chromatography (DHPLC). As a result, we identified 6 DHPLC peak types from MC1R gene and SRY gene analysis. DHPLC method showed more sensitive than RFLP method for DNA fragments analysis. Therefore, DHPLC method can apply to identify for Hanwoo, Holstein and mixed beef.

Discrimination of Hanwoo from Holstein and Mixed Beef by DHPLC

Chungnam Animal Science Center GenetBio, Konyang Univ, Nonsan 320-711, Korea(Received April 8, 2008)요약. 결핵은 전 세계적으로 심각한 공중보건문제로 남아있다. 최근에는 결핵의 발병률이 증가하고 있는추세이며 이에 따른 결핵의 정확한 조기치료와 심각한 부작용, 그리고 전염을 방지하기 위해서는 신속하고 정확한 결핵의 진단이 절실하게 필요하다. 본 연구에서는 결핵유전자를 빠르게 검출하는데 있어서 등온증폭법이 가지고 있는 높은 특이성과 신속성에 대하여 평가하였다. 결핵 DNA의 순차적 10배위 정량희석 DNA를 사용하였으며 일반PCR 방법과 등온증폭법의 실험방법의 검출한계, 민감성, 특이성, 재현성을 비교하였다. 그 결과 등온 증폭법은 빠른 증폭 시간과 높은 민감성, 높은 특이성을 가지고 있었으며 병원이나 연구실, 진단 검사실 등에서 결핵유전자를 빠르고 정확하게 진단할 수 있는 유용한 방법이 될 수 있을 것이다. 주제어: 결핵, PCR, 등온증폭법, FIP(Forward Inner Primer), BIP(Back Inner Primer) ABSTRACT. Mycobacterium tuberculosis (MTB) remains a major worldwide public health problem. In recent years,the incidence of MTB has been rising. Rapid and reliable diagnosis of Mycobacterium tuberculosis is essential to initiatecorrect treatment, avoid severe complications, and prevent transmission. LAMP was used to develop a rapid and sensitivelaboratory diagnostic system for the MTB. In this research, the loop-mediated isothermal amplification method (LAMP)that amplifies DNA with high specificity and rapidity at an isothermal condition was evaluated for rapid detection ofMTB. Undiluted DNA (2.10 × 10

Detection of Mycobacterium Tuberculosis by Loop-Mediated Isothermal Amplification Assay

Staphylococcus aureus is the most important pathogen in nosocomial infections, including bloodstream infections. Prompt identification of S. aureus from blood cultures and detection of methicillin resistance are essential in cases of suspected sepsis. We have studied a new method for the sequence-specific visual detection of minute amounts of nucleic acids using intercalating reaction by addition of SYBR Green to amplicons of LAMP, and it's a unique gene amplification method in which DNA can be isothermally amplified using only one enzyme. Staphylococcus-LAMP, which targets the spa gene, encoding S.aureus-specific protein A, and the mecA gene, encoding penicillin-binding protein-2' for methicillin resistance, detected MRSA and MRSE. In this study, by using LAMP assay, I detected for Staphylococcus aureus and Staphylococcus epidermidis concentration in the clinical sample. The detection of Staphylococcus aureus and Staphylococcus epidermidis was tested by using serial 10-fold dilutions standard solution. I have accurate detected the limit of detection, sensitity, specificity and reproducibility of the assay. The Bio-chip based LAMP assay allowed easy, rapid, accurate and sensitive detection of infection with Staphylococcus and especially applicable in a resource-limited situation.

/pdf/detection-for-methicillin-resistant-staphylococcus-aureus-in-375iqmwk7u.pdf

Detection for Methicillin Resistant Staphylococcus aureus in Using Bio-Chip Based Loop Mediated Isothermal Amplification Assay

Deparment of Chemistry, School of Advanced Science and Basic Science Research Institute, Dankook University,Cheonan 330-714, Korea(Received August 28, 2007)요약 MODY는 일반적으로 제 2 형 당뇨의 특수형으로 25세 이전에 발병하며 상염색체 우성으로 유전되며 인슐린 분비장애를 특징으로 한다 GCK와 HNF-1α의 유전자 돌연변이는 제 2 형 당뇨에서 가장 큰 원인중에 하나이다 따라서 이들 유전자다형성의 연관성을 다양한 분석 방법으로 연구할 필요성이 대두되고 있다GCK와 HNF-1α 유전자의 exon과 exon에 근접한 intron을 실험하였고, 또한 promotor까지 PCR-DHPLC(Polymerase Chain Reaction - Denaturing High Performance Liquid Chromatography) 방법과 direct sequencing 방법을 사용하였다 시료는 11명의 환자와 20명의 정상인에서 DNA를 추출하였고 GCK와 HNF-1α의 단일 염기다형성을 PCR-DHPLC 방법으로 확인하였다 결과적으로 GCK 유전자에서는 1개(R135G)를 검출하였고 HNF-1α유전자에서는 2개(I27L, S487N)를 검출하였으며 intron 8에서도 돌연변이를 검출할 수 있었다주제어: MODY, GCK, HNF-1α, DHPLCABSTRACT Maturity-onset diabetes of the young (MODY) is a genetically heterogeneous subtype of Type 2 dia-betes characterized (non-insulin-dependent) by early onset, usually before 25 years of age, autosomal dominant inher-itance and a primary defect in insulin secretion Mutations in the glucokinase (GCK) and hepatocyte nuclear factor(HNF)-1α genes are the major causes of monogenic forms of Type 2 diabetes mellitus Therefore it is need to study rela-tion with these polymorphisms by diverse analysis methods The promotor and coding regions inclusive intron exonboundaries of the GCK, HNF-1α genes were examined by PCR-DHPLC (Polymerase Chain Reaction - Denaturing HighPerformance Liquid Chromatography) and direct sequencing We extracted DNA from 11 patients and 20 normals Thenwe confirmed a single-nucleotide polymorphism using PCR-DHPLC As results, we identified one mutation (R135G)in GCK gene and two mutations (I27L, S487N) in HNF-1a and at the same time detected mutation in intron 8Keywords: MODY, GCK, HNF-1α, DHPLC

/pdf/analysis-of-the-gck-and-hnf-1a-gene-polymorphism-in-korean-14bzlhe5r8.pdf

Analysis of the GCK and HNF-1α Gene Polymorphism in Korean Type 2 Diabetic Patients by PCR-DHPLC

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