{"title":"用聚合酶链反应扩增HIV-1前病毒DNA的相对定量","authors":"AAM Jiman-Fatani, PJ Vallely, DJ Morris","doi":"10.1016/0888-0786(94)90034-5","DOIUrl":null,"url":null,"abstract":"<div><p>A 141-base pair fragment of human immunodeficiency virus-1 (HIV-1) DNA was amplified using the polymerase chain reaction (PCR). The products were slot-blotted onto a nitrocellulose membrane, revealed with a digoxigenin-labelled probe and quantitated by scanning densitometry. This method for the relative quantitation of HIV-1 DNA achieved reliable results and avoided the use of radioisotopes and the electrophoretic transfer of DNA. Testing of serial dilutions of HIV-1 extracted from infected cells revealed smooth titration curves. A reproducible increase in peripheral blood HIV-1 DNA was documented in a haemophilia patient during disease progression.</p></div>","PeriodicalId":101161,"journal":{"name":"Serodiagnosis and Immunotherapy in Infectious Disease","volume":"6 4","pages":"Pages 209-212"},"PeriodicalIF":0.0000,"publicationDate":"1994-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0888-0786(94)90034-5","citationCount":"0","resultStr":"{\"title\":\"Relative quantitation of HIV-1 proviral DNA amplified using the polymerase chain reaction\",\"authors\":\"AAM Jiman-Fatani, PJ Vallely, DJ Morris\",\"doi\":\"10.1016/0888-0786(94)90034-5\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>A 141-base pair fragment of human immunodeficiency virus-1 (HIV-1) DNA was amplified using the polymerase chain reaction (PCR). The products were slot-blotted onto a nitrocellulose membrane, revealed with a digoxigenin-labelled probe and quantitated by scanning densitometry. This method for the relative quantitation of HIV-1 DNA achieved reliable results and avoided the use of radioisotopes and the electrophoretic transfer of DNA. Testing of serial dilutions of HIV-1 extracted from infected cells revealed smooth titration curves. A reproducible increase in peripheral blood HIV-1 DNA was documented in a haemophilia patient during disease progression.</p></div>\",\"PeriodicalId\":101161,\"journal\":{\"name\":\"Serodiagnosis and Immunotherapy in Infectious Disease\",\"volume\":\"6 4\",\"pages\":\"Pages 209-212\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1994-12-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/0888-0786(94)90034-5\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Serodiagnosis and Immunotherapy in Infectious Disease\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/0888078694900345\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Serodiagnosis and Immunotherapy in Infectious Disease","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/0888078694900345","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
利用聚合酶链反应(PCR)扩增了人类免疫缺陷病毒-1 (HIV-1) DNA 141碱基对片段。将产物在硝化纤维素膜上进行槽印迹，用地高辛标记探针显示，并通过扫描密度测定法定量。该方法对HIV-1 DNA的相对定量获得了可靠的结果，避免了使用放射性同位素和DNA的电泳转移。从感染细胞中提取的HIV-1的连续稀释测试显示平滑的滴定曲线。一名血友病患者在疾病进展期间外周血HIV-1 DNA可重复增加。
Relative quantitation of HIV-1 proviral DNA amplified using the polymerase chain reaction
A 141-base pair fragment of human immunodeficiency virus-1 (HIV-1) DNA was amplified using the polymerase chain reaction (PCR). The products were slot-blotted onto a nitrocellulose membrane, revealed with a digoxigenin-labelled probe and quantitated by scanning densitometry. This method for the relative quantitation of HIV-1 DNA achieved reliable results and avoided the use of radioisotopes and the electrophoretic transfer of DNA. Testing of serial dilutions of HIV-1 extracted from infected cells revealed smooth titration curves. A reproducible increase in peripheral blood HIV-1 DNA was documented in a haemophilia patient during disease progression.
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