{"title":"SYBR Green I、SYBR Safe和Calcein三种染料环介导等温扩增检测人乳头瘤病毒16的敏感性评价","authors":"Nuur Ezzatyhusna, Nurul Izzati, Siti Suraiya, Mn","doi":"10.17576/MH.2017.1202.07","DOIUrl":null,"url":null,"abstract":"Loop-mediated isothermal amplification (LAMP) is a gene amplification technique whereby the amplification products are commonly visualized as turbidity by naked eye in the presence of magnesium pyrophosphate precipitation. An appropriate intercalating dye is important as it could increase the sensitivity and reduce the false positive and false negative results for the detection. The study aimed to compare the performance of three different intercalating dyes; SYBR Green I, SYBR Safe and calcein-based dyes in HPV-16 LAMP assay by naked-eye visualization, gel electrophoresis and real-time monitoring. The LAMP assay was carried out using a Loopamp DNA amplification kit in 25 μl volumes. The reaction mixture was incubated at 60","PeriodicalId":53946,"journal":{"name":"Medicine and Health","volume":"123 1","pages":"220-229"},"PeriodicalIF":0.1000,"publicationDate":"2017-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"Sensitivity Evaluation of SYBR Green I, SYBR Safe and Calcein Dyes for Detection of Human Papillomavirus 16 by Loop-Mediated Isothermal Amplification\",\"authors\":\"Nuur Ezzatyhusna, Nurul Izzati, Siti Suraiya, Mn\",\"doi\":\"10.17576/MH.2017.1202.07\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Loop-mediated isothermal amplification (LAMP) is a gene amplification technique whereby the amplification products are commonly visualized as turbidity by naked eye in the presence of magnesium pyrophosphate precipitation. An appropriate intercalating dye is important as it could increase the sensitivity and reduce the false positive and false negative results for the detection. The study aimed to compare the performance of three different intercalating dyes; SYBR Green I, SYBR Safe and calcein-based dyes in HPV-16 LAMP assay by naked-eye visualization, gel electrophoresis and real-time monitoring. The LAMP assay was carried out using a Loopamp DNA amplification kit in 25 μl volumes. The reaction mixture was incubated at 60\",\"PeriodicalId\":53946,\"journal\":{\"name\":\"Medicine and Health\",\"volume\":\"123 1\",\"pages\":\"220-229\"},\"PeriodicalIF\":0.1000,\"publicationDate\":\"2017-12-22\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Medicine and Health\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.17576/MH.2017.1202.07\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"MEDICINE, GENERAL & INTERNAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Medicine and Health","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.17576/MH.2017.1202.07","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"MEDICINE, GENERAL & INTERNAL","Score":null,"Total":0}
引用次数: 1
摘要
环介导等温扩增(LAMP)是一种基因扩增技术，其扩增产物通常在焦磷酸镁沉淀存在下以肉眼可见为浊度。适当的插层染料是重要的，因为它可以提高灵敏度，减少假阳性和假阴性检测结果。本研究旨在比较三种不同插层染料的性能;SYBR Green I、SYBR Safe和calcein基染料在HPV-16 LAMP检测中的裸眼可视化、凝胶电泳和实时监测。LAMP检测采用Loopamp DNA扩增试剂盒，体积为25 μl。反应混合物在60℃下孵育
Sensitivity Evaluation of SYBR Green I, SYBR Safe and Calcein Dyes for Detection of Human Papillomavirus 16 by Loop-Mediated Isothermal Amplification
Loop-mediated isothermal amplification (LAMP) is a gene amplification technique whereby the amplification products are commonly visualized as turbidity by naked eye in the presence of magnesium pyrophosphate precipitation. An appropriate intercalating dye is important as it could increase the sensitivity and reduce the false positive and false negative results for the detection. The study aimed to compare the performance of three different intercalating dyes; SYBR Green I, SYBR Safe and calcein-based dyes in HPV-16 LAMP assay by naked-eye visualization, gel electrophoresis and real-time monitoring. The LAMP assay was carried out using a Loopamp DNA amplification kit in 25 μl volumes. The reaction mixture was incubated at 60
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