PERFORMANCE OF LOOP-MEDIATED ISOTHERMAL AMPLIFICATION FOR PATHOGEN DETECTION

• Main Authors: Po-Ching Chen, 陳柏卿
• Other Authors: Chi-Yang Yu
• Format: Others
• Language: en_US
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/16612531153331864482

碩士 === 大同大學 === 生物工程學系(所) === 96 === This thesis reviewed the articles that applied loop-mediated isothermal amplification (LAMP) to the detection of pathogens, and compared the specificities and sensitivities of LAMP to PCR, reverse transcription PCR, nested PCR, as well as real-time PCR. The LAMP reaction requires a set of four specially designed primers and a DNA polymerase, under isothermal conditions (60-65℃), can rapidly amplify target DNA. The specificity of LAMP is extremely high because the set of four specially designed primers recognize six distinct target DNA sequences which play the role of initiating the follow-up reactions. Comparing to PCR, LAMP has short reaction time because it doesn’t need the thermal cycling steps. Furthermore, simple equipments such as a water bath or heat block are sufficient for the DNA amplification. LAMP reaction yields a large amount of the by-product pyrophosphate ion, which reacts with magnesium ions in the reaction to form white magnesium pyrophosphate precipitate. As a result, real-time monitoring of the LAMP reaction can be achieved by the naked eye or by real-time measurement of turbidity. In addition, the sensitivity of staining LAMP amplicons with fluorescent intercalating dye such as SYBR green equals that of gel electrophoresis method. Since the first report of LAMP in 2000, applications of LAMP for the detection of pathogens of human, animals, agriculture and fishing industry diseases were reported by more and more publications. After reviewing selected articles, I conclude that LAMP is highly specific and the sensitivity of LAMP is superior to PCR, and is comparable to nested PCR and real-time PCR.