Dengue virus,a member of the Flavivirus genus in the Flaviviridae family,has four serotypes(DEN-1 to DEN-4),and is responsible for both mild and severe dengue fever(dengue hemorrhagic fever and dengue shock syndrome)[...Dengue virus,a member of the Flavivirus genus in the Flaviviridae family,has four serotypes(DEN-1 to DEN-4),and is responsible for both mild and severe dengue fever(dengue hemorrhagic fever and dengue shock syndrome)[1].Dengue cases are under reported in India and the cases are treated clinically based on the clinical triage of symptoms[2].Nonstructural protein(NS1)antigen is a highly conserved glycoprotein found in Flavivirus group such as Dengue,Zika,and Japanese encephalitis virus.Simultaneous detection of NS1 antigen,IgM and IgG antibody in a single cassette is used for testing primary and secondary dengue infection.Early diagnosis of disease progression associated with severe dengue and accessibility to appropriate medical care reduces severe dengue fatality rates to less than 1%.This retrospective study,conducted from January 2021 to December 2021 in the tertiary care hospital,South India,was aimed to determine the seroprevalence of dengue adult patients with acute febrile illness.展开更多
目的建立SYBR Green I荧光染料实时定量RT-PCR方法,测定猴免疫缺陷病毒(SIV)RNA拷贝数。方法巢式RT-PCR扩增SIV病毒RNAgag基因上1360-1837之间的长度为477 bp的片段,将该片段克隆到pGEMT载体上,构建pGEM-SIVgag477质粒。该质粒经限制性...目的建立SYBR Green I荧光染料实时定量RT-PCR方法,测定猴免疫缺陷病毒(SIV)RNA拷贝数。方法巢式RT-PCR扩增SIV病毒RNAgag基因上1360-1837之间的长度为477 bp的片段,将该片段克隆到pGEMT载体上,构建pGEM-SIVgag477质粒。该质粒经限制性内切酶NotⅠ酶切后,进行体外转录,转录出的RNA产物(RS)纯化后10倍系列稀释,作出标准曲线,作为SIV病毒RNA荧光定量检测的外标准品。结果应用Qiagen公司QuantiTect SYBR GREEN RT-PCR Kit,该标准品可精确定量到100 copies/μL。结论制备的RS外标准品纯度高,SYBR Green I荧光染料实时定量RT-PCR法特异性、敏感性高,稳定性好,可用于定量测定猴免疫缺陷病毒(SIV)RNA拷贝数。展开更多
文摘Dengue virus,a member of the Flavivirus genus in the Flaviviridae family,has four serotypes(DEN-1 to DEN-4),and is responsible for both mild and severe dengue fever(dengue hemorrhagic fever and dengue shock syndrome)[1].Dengue cases are under reported in India and the cases are treated clinically based on the clinical triage of symptoms[2].Nonstructural protein(NS1)antigen is a highly conserved glycoprotein found in Flavivirus group such as Dengue,Zika,and Japanese encephalitis virus.Simultaneous detection of NS1 antigen,IgM and IgG antibody in a single cassette is used for testing primary and secondary dengue infection.Early diagnosis of disease progression associated with severe dengue and accessibility to appropriate medical care reduces severe dengue fatality rates to less than 1%.This retrospective study,conducted from January 2021 to December 2021 in the tertiary care hospital,South India,was aimed to determine the seroprevalence of dengue adult patients with acute febrile illness.
文摘目的建立SYBR Green I荧光染料实时定量RT-PCR方法,测定猴免疫缺陷病毒(SIV)RNA拷贝数。方法巢式RT-PCR扩增SIV病毒RNAgag基因上1360-1837之间的长度为477 bp的片段,将该片段克隆到pGEMT载体上,构建pGEM-SIVgag477质粒。该质粒经限制性内切酶NotⅠ酶切后,进行体外转录,转录出的RNA产物(RS)纯化后10倍系列稀释,作出标准曲线,作为SIV病毒RNA荧光定量检测的外标准品。结果应用Qiagen公司QuantiTect SYBR GREEN RT-PCR Kit,该标准品可精确定量到100 copies/μL。结论制备的RS外标准品纯度高,SYBR Green I荧光染料实时定量RT-PCR法特异性、敏感性高,稳定性好,可用于定量测定猴免疫缺陷病毒(SIV)RNA拷贝数。