{"title":"[TapMan实时RT-PCR检测HHV-6方法的建立及临床应用]。","authors":"Qian-Qian Chen, Bing Zhang, Zhi-Ping Xie, Jin-Song Li, Han-Chun Gao, Ni-Guang Xiao, Le-Yun Xie, Tian Yu, Sai-Zhen Zeng, Ping Gong, Zhao-Jun Duan","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To establish a rapid, sensitive and specific real-time PCR method for detection of Human Herpesvirus-6 (HHV-6).</p><p><strong>Methods: </strong>According to the reference, a pair of primers and a probe were designed located in U65-66 gene and to set up the standards. We established a real-time RT-PCR method for detection of HHV-6, and to verify the specificity, sensitivity, reproducibility.</p><p><strong>Results: </strong>The correlation coefficient was 0.999, E = 97.9%, the coefficient of variation values of Ct were 0.61% and 3.13% in real-time PCR assay for inter and intra assay, respectively. The results of all viruses were negative except of HHV-6 for the assay. The quantitative detection limit of the assay was 3 x 10(0) copies/microl.</p><p><strong>Conclusion: </strong>The real-time PCR assay is highly specific, sensitive and reproducible, which can be used to quatitative detecting clinical samples.</p>","PeriodicalId":70973,"journal":{"name":"中华实验和临床病毒学杂志","volume":"27 2","pages":"144-6"},"PeriodicalIF":0.0000,"publicationDate":"2013-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Establishment and clinical application of TapMan real-time RT-PCR method for detection of HHV-6].\",\"authors\":\"Qian-Qian Chen, Bing Zhang, Zhi-Ping Xie, Jin-Song Li, Han-Chun Gao, Ni-Guang Xiao, Le-Yun Xie, Tian Yu, Sai-Zhen Zeng, Ping Gong, Zhao-Jun Duan\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>To establish a rapid, sensitive and specific real-time PCR method for detection of Human Herpesvirus-6 (HHV-6).</p><p><strong>Methods: </strong>According to the reference, a pair of primers and a probe were designed located in U65-66 gene and to set up the standards. We established a real-time RT-PCR method for detection of HHV-6, and to verify the specificity, sensitivity, reproducibility.</p><p><strong>Results: </strong>The correlation coefficient was 0.999, E = 97.9%, the coefficient of variation values of Ct were 0.61% and 3.13% in real-time PCR assay for inter and intra assay, respectively. The results of all viruses were negative except of HHV-6 for the assay. The quantitative detection limit of the assay was 3 x 10(0) copies/microl.</p><p><strong>Conclusion: </strong>The real-time PCR assay is highly specific, sensitive and reproducible, which can be used to quatitative detecting clinical samples.</p>\",\"PeriodicalId\":70973,\"journal\":{\"name\":\"中华实验和临床病毒学杂志\",\"volume\":\"27 2\",\"pages\":\"144-6\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2013-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"中华实验和临床病毒学杂志\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"中华实验和临床病毒学杂志","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
[Establishment and clinical application of TapMan real-time RT-PCR method for detection of HHV-6].
Objective: To establish a rapid, sensitive and specific real-time PCR method for detection of Human Herpesvirus-6 (HHV-6).
Methods: According to the reference, a pair of primers and a probe were designed located in U65-66 gene and to set up the standards. We established a real-time RT-PCR method for detection of HHV-6, and to verify the specificity, sensitivity, reproducibility.
Results: The correlation coefficient was 0.999, E = 97.9%, the coefficient of variation values of Ct were 0.61% and 3.13% in real-time PCR assay for inter and intra assay, respectively. The results of all viruses were negative except of HHV-6 for the assay. The quantitative detection limit of the assay was 3 x 10(0) copies/microl.
Conclusion: The real-time PCR assay is highly specific, sensitive and reproducible, which can be used to quatitative detecting clinical samples.
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