Objective:To report high co-positivity of anti-dengue virus(DV)and anti-Japanese encephalitis virus(JEV)IgM in an area endemic for both the viruses and to discuss the possibilities of coinfection.Methods:Serum samples...Objective:To report high co-positivity of anti-dengue virus(DV)and anti-Japanese encephalitis virus(JEV)IgM in an area endemic for both the viruses and to discuss the possibilities of coinfection.Methods:Serum samples from the patients who presented with fever,suspected central nervous system infection and thrombocytopenia,were tested for anti-DV IgM and antiJEV IgM antibodies.Conventional reverse transcriptase polymerase chain reaction was done for detection of DV RNA and JEV RNA.Results:Of 1 410 patient sera tested for anti-DV and antiJEV antibodies,129(9.14%)were co-positive for both.This co-positivity was observed only in those months when anli-JEV IgM positivily was high.Tilers of both anli-DV IgM and anti-JEV IgM were high in most of the co-positive eases.Among these 129 co-positive cases,76 were lesled by conventional reverse Iranscriplase polymerase chain reaction for both flaviviruses,of which eight cases were co-positive for DV and JEV.Conclusions:Co-infection with more than one fluvivirus species can occur in hyperendemic areas.展开更多
Viral nervous necrosis （VNN） causes high mortality in marine fish, especially in the grouper, worldwide and in China. Since there is no effective vaccine or drug to deal with VNN, early detection and prevention is i...Viral nervous necrosis （VNN） causes high mortality in marine fish, especially in the grouper, worldwide and in China. Since there is no effective vaccine or drug to deal with VNN, early detection and prevention is important to block its outbreak. In this study, a reverse transcription-polymerase chain reaction （RT-PCR） was developed for the rapid, convenient, and sensitive detection of the VNN pathogen, nervous necro- sis virus （NNV）, in the grouper. The whole process was completed within 3.5 h from the RNA extraction to PCR product visualization. The detection limit of this method was 200 copies of NNV RNA standard, which corresponded to 200 copies of virus particles. This RT-PCR method was specific to the NNV detection with no cross-reactivity to other fish viral disease pathogens, such as infectious pancreatic necrosis virus （IPNV）, infectious hematopoietic necrosis virus （IHNV）, spring viraemia of carp virus （SVCV）, epizootic haematopoietic necrosis virus （EHNV）, and large yellow croaker iridovirus （LYC1V）. With this method, the orange-spotted grouper （Epinephelus coioides） fry from hatcheries with or without incidence of the VN- N epidemic in Fujian Province were detected. The results showed that all or 93% of the fry from the two hatcheries with incidence of the epidemic were diagnosed as positive, while 40% or 25% of fry from the t- wo hatcheries without the VNN epidemic were also detected as NNV positive, indicating that this RT-PCR method can be used for rapid, sensitive detection of NNV infection and applied in the VNN epidemic alert.展开更多
The expression of the cytokines IL-2, IL-10, TNF-α and their roles in mice with herpes simplex viral encephalitis （HSE） were studied. By using semiquantitative reverse transcription polymerase chain reaction （RT-...The expression of the cytokines IL-2, IL-10, TNF-α and their roles in mice with herpes simplex viral encephalitis （HSE） were studied. By using semiquantitative reverse transcription polymerase chain reaction （RT-PCR）, the expressions of IL-2, IL-10 and TNF-α mRNA in control group, HSE group and acyclovir （ACV）-treated group were detected and the pathological changes of brain were observed. It was found that after HSV1 infection, the cerebral lesions of haemorrhage and necrosis in mice were observed under the microscopy, and the levels of IL-2, IL-10 and TNF-α were increased remarkably. After treatment with ACV after HSV1 infection, the cerebral lesions in mice were improved, the level of IL-2 maintained stable, IL-10 was increased consistently, and TNF-α was decreased significantly as compared with those in HSE group. In acute HSE, many cytokines are upregulated, including IL-2, IL-10 and TNF-α to eliminate virus and TH1 type response is dominant. In convalescence, there is a shift in the cytokine expression profile from TH1 profile to TH2 profile and the shift can inhibit the overexpression of immune response in animals. ACV has remarkable effects in the treatment of HSE.展开更多
Forty-two patients with hepatocellular carcinoma (HCC) were examined for hepatitis C virus (HCV) RNA in liver tissues by reverse transcription-polymerase chain reaction (RT-PCR) with primers deduced from 5'-non-co...Forty-two patients with hepatocellular carcinoma (HCC) were examined for hepatitis C virus (HCV) RNA in liver tissues by reverse transcription-polymerase chain reaction (RT-PCR) with primers deduced from 5'-non-coding region (5'-NCR). HCV liver samples of展开更多
根据大蒜普通潜隐病毒(Garlic common latent virus,GCLV)的外壳蛋白区域的保守序列设计合成一对寡核苷酸引物,以带毒植物的总RNA为模板,进行反转录和PCR扩增,通过反应体系和反应程序的建立与优化,扩增得到长300 bp的目的片段,并将目的...根据大蒜普通潜隐病毒(Garlic common latent virus,GCLV)的外壳蛋白区域的保守序列设计合成一对寡核苷酸引物,以带毒植物的总RNA为模板,进行反转录和PCR扩增,通过反应体系和反应程序的建立与优化,扩增得到长300 bp的目的片段,并将目的片段转入大肠杆菌进行了克隆和序列测定。测序结果与GenBank中其他GCLV相应区域的序列同源性最高达98%。并对其检测的特异性和灵敏度进行了验证,从而建立了快速、灵敏、特异性强的GCLV分子生物学检测方法。展开更多
基金Financial support from Indian Council of Medical Research.New DelhiCouncil of Scientific,Industrial Research,New Delhi
文摘Objective:To report high co-positivity of anti-dengue virus(DV)and anti-Japanese encephalitis virus(JEV)IgM in an area endemic for both the viruses and to discuss the possibilities of coinfection.Methods:Serum samples from the patients who presented with fever,suspected central nervous system infection and thrombocytopenia,were tested for anti-DV IgM and antiJEV IgM antibodies.Conventional reverse transcriptase polymerase chain reaction was done for detection of DV RNA and JEV RNA.Results:Of 1 410 patient sera tested for anti-DV and antiJEV antibodies,129(9.14%)were co-positive for both.This co-positivity was observed only in those months when anli-JEV IgM positivily was high.Tilers of both anli-DV IgM and anti-JEV IgM were high in most of the co-positive eases.Among these 129 co-positive cases,76 were lesled by conventional reverse Iranscriplase polymerase chain reaction for both flaviviruses,of which eight cases were co-positive for DV and JEV.Conclusions:Co-infection with more than one fluvivirus species can occur in hyperendemic areas.
基金The National Basic Research Program under contract No.2012CB114402the National High Technology Research and Development Program of China (863 Program) under contract No.2012AA092202Scientific Research Project of Marine Public Welfare Industry of China under contract No.200905020-07
文摘Viral nervous necrosis （VNN） causes high mortality in marine fish, especially in the grouper, worldwide and in China. Since there is no effective vaccine or drug to deal with VNN, early detection and prevention is important to block its outbreak. In this study, a reverse transcription-polymerase chain reaction （RT-PCR） was developed for the rapid, convenient, and sensitive detection of the VNN pathogen, nervous necro- sis virus （NNV）, in the grouper. The whole process was completed within 3.5 h from the RNA extraction to PCR product visualization. The detection limit of this method was 200 copies of NNV RNA standard, which corresponded to 200 copies of virus particles. This RT-PCR method was specific to the NNV detection with no cross-reactivity to other fish viral disease pathogens, such as infectious pancreatic necrosis virus （IPNV）, infectious hematopoietic necrosis virus （IHNV）, spring viraemia of carp virus （SVCV）, epizootic haematopoietic necrosis virus （EHNV）, and large yellow croaker iridovirus （LYC1V）. With this method, the orange-spotted grouper （Epinephelus coioides） fry from hatcheries with or without incidence of the VN- N epidemic in Fujian Province were detected. The results showed that all or 93% of the fry from the two hatcheries with incidence of the epidemic were diagnosed as positive, while 40% or 25% of fry from the t- wo hatcheries without the VNN epidemic were also detected as NNV positive, indicating that this RT-PCR method can be used for rapid, sensitive detection of NNV infection and applied in the VNN epidemic alert.
文摘The expression of the cytokines IL-2, IL-10, TNF-α and their roles in mice with herpes simplex viral encephalitis （HSE） were studied. By using semiquantitative reverse transcription polymerase chain reaction （RT-PCR）, the expressions of IL-2, IL-10 and TNF-α mRNA in control group, HSE group and acyclovir （ACV）-treated group were detected and the pathological changes of brain were observed. It was found that after HSV1 infection, the cerebral lesions of haemorrhage and necrosis in mice were observed under the microscopy, and the levels of IL-2, IL-10 and TNF-α were increased remarkably. After treatment with ACV after HSV1 infection, the cerebral lesions in mice were improved, the level of IL-2 maintained stable, IL-10 was increased consistently, and TNF-α was decreased significantly as compared with those in HSE group. In acute HSE, many cytokines are upregulated, including IL-2, IL-10 and TNF-α to eliminate virus and TH1 type response is dominant. In convalescence, there is a shift in the cytokine expression profile from TH1 profile to TH2 profile and the shift can inhibit the overexpression of immune response in animals. ACV has remarkable effects in the treatment of HSE.
文摘Forty-two patients with hepatocellular carcinoma (HCC) were examined for hepatitis C virus (HCV) RNA in liver tissues by reverse transcription-polymerase chain reaction (RT-PCR) with primers deduced from 5'-non-coding region (5'-NCR). HCV liver samples of
文摘根据大蒜普通潜隐病毒(Garlic common latent virus,GCLV)的外壳蛋白区域的保守序列设计合成一对寡核苷酸引物,以带毒植物的总RNA为模板,进行反转录和PCR扩增,通过反应体系和反应程序的建立与优化,扩增得到长300 bp的目的片段,并将目的片段转入大肠杆菌进行了克隆和序列测定。测序结果与GenBank中其他GCLV相应区域的序列同源性最高达98%。并对其检测的特异性和灵敏度进行了验证,从而建立了快速、灵敏、特异性强的GCLV分子生物学检测方法。
