Objectives: To compare multiplex fluorescent PCRwith serum type-specific antibody detection in thediagnosis of herpes simplex virus (HSV) infection andto evaluate its significance in the diagnosis of genitalherpes.Met...Objectives: To compare multiplex fluorescent PCRwith serum type-specific antibody detection in thediagnosis of herpes simplex virus (HSV) infection andto evaluate its significance in the diagnosis of genitalherpes.Methods: We detected HSV infection in 121 speci-mens collected from patients with genital herpesusing both multiplex fluorescent PCR and serum type-specific antibody detection. HSV viral isolation wasused as the standard control.Results: When compared with the viral isolation, thesensitivity and specificity for multiplex fluorescentPCR were 100% and 88.89%, respectively afterdiscrepant analysis. The sensitivity and specificity fortype-specific antibody detection was 77.68 % and77.78 %, respectively. However, the type-specificantibody detected HSV in two asymptomatic patientswhile the multiplex fluorescent PCR couldn’t detectany HSV DNA from those specimens.Conclusions: Multiplex fluorescent PCR is a verysensitive and specific method for detection and typingof HSV in the lesion of genital herpes, it failed todetect HSV DNA from the asymptomatic patients.Serum type-specific antibody detection was a lesssensitive and specific test but could detect the specificantibody from some asymptomatic patients. Thecombination of these two techniques would allow rapid,sensitive and accurate detection and typing of HSVand help clinical diagnosis and epidemiologic survey-ing of genital herpes.展开更多
Grass carp reovirus （GCRV）, a double stranded RNA virus that infects aquatic animals, often with disastrous effects, belongs to the genus Aquareovirus and family Reoviridea. Similar to other reoviruses, genome repli...Grass carp reovirus （GCRV）, a double stranded RNA virus that infects aquatic animals, often with disastrous effects, belongs to the genus Aquareovirus and family Reoviridea. Similar to other reoviruses, genome replication of GCRV in infected cells occurs in cytoplasmic inclusion bodies, also called viral factories. Sequences analysis revealed the nonstmctural protein NS80, encoded by GCRV segment 4, has a high similarity with μNS in MRV（Mammalian orthoreovimses）, which may be associated with viral factory formation. To understand the function of the μNS80 protein in virus replication, the initial expression and identification of the immunogenicity of the GCRV NS80 protein inclusion forming-related region （335-742） was investigated in this study. It is shown that the over-expressed fusion protein was produced by inducing with IPTG at 28℃. In addition, serum specific rabbit antibody was obtained by using super purified recombinant NS80（335-742） protein as antigen. Moreover, the expressed protein was able to bind to anti-his-tag monoclonal antibody （mouse） and NS80〈335.742） specific rabbit antibody. Further western blot analysis indicates that the antiserum could detect NS80 or NS80C protein expression in GCRV infected cells. This data provides a foundation for further investigation of the role of NS80 in viral inclusion formation and virion assembly.展开更多
Hepatitis B virus (HBV) is a common viral pathogen that causes a substantial health burden worldwide. Remarkable progress has been made in our understanding of the natural stages of chronic HBV infection. A dynamic ba...Hepatitis B virus (HBV) is a common viral pathogen that causes a substantial health burden worldwide. Remarkable progress has been made in our understanding of the natural stages of chronic HBV infection. A dynamic balance between viral replication and host immune response is pivotal to the pathogenesis of liver disease. Knowledge of the HBV genome organization and replication cycle can unravel HBV genotypes and molecular variants, which contribute to the heterogeneity in outcome of chronic HBV infection. Most HBV infections are spontaneously resolved in immunocompetent adults, whereas they become chronic in most neonates and infants at a great risk of developing complications such as cirrhosis and hepatocellular carcinoma (HCC). Those with chronic HBV infection may present in one of the four phases of infection: immune tolerance, immune clearance [hepatitis B eantigen (HBeAg)-positive chronic hepatitis B (CHB)], inactive carrier state, and reactivation (HBeAg-negative CHB). Understanding the dynamic nature of chronic HBV infection is crucial in the management of HBV carriers. Long-term monitoring and optimal timing of antiviral therapy for chronic HBV infection help to prevent progression of HBV-related liver disease to its later stage, particularly in patients with higher risk markers of HCC, such as serum DNA concentration, HBeAg status, serum aminotransferase, HBV genotypes, and pre-core or core mutants.展开更多
Viral nervous necrosis （VNN） has been reported to infect larvae and juvenile of humpback grouper, and until now, this virus is one of main problem for humpback grouper. Co-agglutination test proved to be a simple, r...Viral nervous necrosis （VNN） has been reported to infect larvae and juvenile of humpback grouper, and until now, this virus is one of main problem for humpback grouper. Co-agglutination test proved to be a simple, rapid and reliable diagnostic test suitable for use in the field or laboratory without any special apparatus. The study aimed to study application of a co-agglutination test using Staphylococci sensitized with specific antibody for the diagnostic of VNN in grouper. First, brain and eye organ samples from diseased fish are homogenized with phosphate buffer saline （PBS） of pH 7.2. Then, the supernatant is collected after centrifugation at 8,000 rpm for 20 min, and finally one drop of the supematant and one drop of anti VNN antibody sensitized Staphylococci suspension are mixed on a glass slid and observation of the agglutination is performed after 5-10 min. The result shows that co-agglutination technique detects positive VNN in the brain and eye samples. The co-agglutination technique may provide valid result in a very short time as compared with complex method, such as enzyme-linked immunosorbant essay （ELISA） and fluorescient antibody technique （FAT） that require a high cost. Thus, this test can detect VNN faster, simple and more economic.展开更多
OBJECTIVE： To test the in vitro antiviral activity of a crude tissue extract （CTE/from the earthworm Eisenia fetida, determine any effective components in the CTE, andelucidate possiblemechanismsofaction. METHODS： ...OBJECTIVE： To test the in vitro antiviral activity of a crude tissue extract （CTE/from the earthworm Eisenia fetida, determine any effective components in the CTE, andelucidate possiblemechanismsofaction. METHODS： A CTE was made by homogenizing earthworms, followed by treatment with ammoni- um sulfate, then thermal denaturation. Inhibition of virus-induced cytopathic effect （CPE） was used to assess antiviral activity. Chromatographic analy- sis was used to identify effective components in the CTE. RESULTS： The CTE inhibited viral CPE at non-cyto- toxic concentrations. Chromatography indicated that antiviral components corresponded to three active peaks indicative of proteases, nucleases and lysozymes. For adenoviruses, reduction in viral ac- tivity occurred for 100 lag/mL CTE. The reduction in adenoviral activity for four fractions was 100%, 91.8%, 86.9%, and 94.7%. For influenza viruses, re- duction in viral activity of 100%, 86.6%, 69.1% and 88.3% was observed for 37 pg/mL CTE. In addition, three active fractions mixture had stronger antiviral activity （98.7% and 96.7%） than three fractions alone.Gel electrophoresis results indicated that nu- cleases from E. fetida could degrade the genome of influenza viruses and adenoviruses. CONCLUSION： The earthworm CTE displayed non-specific antiviral properties, possibly mediated by a combination of proteases, nucleases and lyso- zymes. Nucleases likely participate in the antiviral process, and degrade the genome of the virus thereby preventing further replication.展开更多
文摘Objectives: To compare multiplex fluorescent PCRwith serum type-specific antibody detection in thediagnosis of herpes simplex virus (HSV) infection andto evaluate its significance in the diagnosis of genitalherpes.Methods: We detected HSV infection in 121 speci-mens collected from patients with genital herpesusing both multiplex fluorescent PCR and serum type-specific antibody detection. HSV viral isolation wasused as the standard control.Results: When compared with the viral isolation, thesensitivity and specificity for multiplex fluorescentPCR were 100% and 88.89%, respectively afterdiscrepant analysis. The sensitivity and specificity fortype-specific antibody detection was 77.68 % and77.78 %, respectively. However, the type-specificantibody detected HSV in two asymptomatic patientswhile the multiplex fluorescent PCR couldn’t detectany HSV DNA from those specimens.Conclusions: Multiplex fluorescent PCR is a verysensitive and specific method for detection and typingof HSV in the lesion of genital herpes, it failed todetect HSV DNA from the asymptomatic patients.Serum type-specific antibody detection was a lesssensitive and specific test but could detect the specificantibody from some asymptomatic patients. Thecombination of these two techniques would allow rapid,sensitive and accurate detection and typing of HSVand help clinical diagnosis and epidemiologic survey-ing of genital herpes.
基金National Basic Research Program of China (973 Program, Grant No. 2009CB118701)National Natural Scientific Foundation of China (Grant Nos. 30671615, 30871940)Innovation project of the Chinese Academy of Sciences (Grant No.KSCX2-YW-N-021)
文摘Grass carp reovirus （GCRV）, a double stranded RNA virus that infects aquatic animals, often with disastrous effects, belongs to the genus Aquareovirus and family Reoviridea. Similar to other reoviruses, genome replication of GCRV in infected cells occurs in cytoplasmic inclusion bodies, also called viral factories. Sequences analysis revealed the nonstmctural protein NS80, encoded by GCRV segment 4, has a high similarity with μNS in MRV（Mammalian orthoreovimses）, which may be associated with viral factory formation. To understand the function of the μNS80 protein in virus replication, the initial expression and identification of the immunogenicity of the GCRV NS80 protein inclusion forming-related region （335-742） was investigated in this study. It is shown that the over-expressed fusion protein was produced by inducing with IPTG at 28℃. In addition, serum specific rabbit antibody was obtained by using super purified recombinant NS80（335-742） protein as antigen. Moreover, the expressed protein was able to bind to anti-his-tag monoclonal antibody （mouse） and NS80〈335.742） specific rabbit antibody. Further western blot analysis indicates that the antiserum could detect NS80 or NS80C protein expression in GCRV infected cells. This data provides a foundation for further investigation of the role of NS80 in viral inclusion formation and virion assembly.
基金Supported by Science and Technology Department of Qingdao Government 07-2-1-15-nsh
文摘Hepatitis B virus (HBV) is a common viral pathogen that causes a substantial health burden worldwide. Remarkable progress has been made in our understanding of the natural stages of chronic HBV infection. A dynamic balance between viral replication and host immune response is pivotal to the pathogenesis of liver disease. Knowledge of the HBV genome organization and replication cycle can unravel HBV genotypes and molecular variants, which contribute to the heterogeneity in outcome of chronic HBV infection. Most HBV infections are spontaneously resolved in immunocompetent adults, whereas they become chronic in most neonates and infants at a great risk of developing complications such as cirrhosis and hepatocellular carcinoma (HCC). Those with chronic HBV infection may present in one of the four phases of infection: immune tolerance, immune clearance [hepatitis B eantigen (HBeAg)-positive chronic hepatitis B (CHB)], inactive carrier state, and reactivation (HBeAg-negative CHB). Understanding the dynamic nature of chronic HBV infection is crucial in the management of HBV carriers. Long-term monitoring and optimal timing of antiviral therapy for chronic HBV infection help to prevent progression of HBV-related liver disease to its later stage, particularly in patients with higher risk markers of HCC, such as serum DNA concentration, HBeAg status, serum aminotransferase, HBV genotypes, and pre-core or core mutants.
文摘Viral nervous necrosis （VNN） has been reported to infect larvae and juvenile of humpback grouper, and until now, this virus is one of main problem for humpback grouper. Co-agglutination test proved to be a simple, rapid and reliable diagnostic test suitable for use in the field or laboratory without any special apparatus. The study aimed to study application of a co-agglutination test using Staphylococci sensitized with specific antibody for the diagnostic of VNN in grouper. First, brain and eye organ samples from diseased fish are homogenized with phosphate buffer saline （PBS） of pH 7.2. Then, the supernatant is collected after centrifugation at 8,000 rpm for 20 min, and finally one drop of the supematant and one drop of anti VNN antibody sensitized Staphylococci suspension are mixed on a glass slid and observation of the agglutination is performed after 5-10 min. The result shows that co-agglutination technique detects positive VNN in the brain and eye samples. The co-agglutination technique may provide valid result in a very short time as compared with complex method, such as enzyme-linked immunosorbant essay （ELISA） and fluorescient antibody technique （FAT） that require a high cost. Thus, this test can detect VNN faster, simple and more economic.
文摘OBJECTIVE： To test the in vitro antiviral activity of a crude tissue extract （CTE/from the earthworm Eisenia fetida, determine any effective components in the CTE, andelucidate possiblemechanismsofaction. METHODS： A CTE was made by homogenizing earthworms, followed by treatment with ammoni- um sulfate, then thermal denaturation. Inhibition of virus-induced cytopathic effect （CPE） was used to assess antiviral activity. Chromatographic analy- sis was used to identify effective components in the CTE. RESULTS： The CTE inhibited viral CPE at non-cyto- toxic concentrations. Chromatography indicated that antiviral components corresponded to three active peaks indicative of proteases, nucleases and lysozymes. For adenoviruses, reduction in viral ac- tivity occurred for 100 lag/mL CTE. The reduction in adenoviral activity for four fractions was 100%, 91.8%, 86.9%, and 94.7%. For influenza viruses, re- duction in viral activity of 100%, 86.6%, 69.1% and 88.3% was observed for 37 pg/mL CTE. In addition, three active fractions mixture had stronger antiviral activity （98.7% and 96.7%） than three fractions alone.Gel electrophoresis results indicated that nu- cleases from E. fetida could degrade the genome of influenza viruses and adenoviruses. CONCLUSION： The earthworm CTE displayed non-specific antiviral properties, possibly mediated by a combination of proteases, nucleases and lyso- zymes. Nucleases likely participate in the antiviral process, and degrade the genome of the virus thereby preventing further replication.
