OBJECTIVETo investigate hepatitis E virus (HEV) infection among pigs in Henan province.

METHODSA total of 623 swine sera, collected from 5 districts, were divided into two groups, under 3-month of age and over 3-month of age. They were tested for HEV antigen and antibody by using ELISAs, respectively. The sera positive for HEV antigen were tested for HEV RNA with RT-PCR. The positive products of RT-PCR were cloned and sequenced.

RESULTSThe positive rates of anti-HEV antibody of the groups under 3-month and over 3-month of age were 90.27% and 92.55%, respectively, without statistical difference, while those of HEV antigen were 15.93% and 5.69%, respectively, with significant difference. The positive rates of anti-HEV antibody and HEV antigen were significantly different among different districts. HEV RNA was detectable in 5 of 47 HEV antigen positive samples. The sequence analysis showed that in 4 of 5 specimens the sequence belonged to genotype 4 while in the remaining one the sequence was genotype 1.

CONCLUSIONThe prevalence rate of HEV infection in pigs was high in Henan province and the rate differed in different districts.

Animals ; Antibodies, Viral ; analysis ; immunology ; Antigens, Viral ; analysis ; immunology ; China ; Genotype ; Hepatitis E ; epidemiology ; immunology ; veterinary ; virology ; Hepatitis E virus ; genetics ; immunology ; isolation & purification ; Phylogeny ; RNA, Viral ; analysis ; genetics ; Sequence Analysis, DNA ; Swine ; virology ; Swine Diseases ; epidemiology ; immunology ; virology

This experiment was performed to determine whether cultured rabbit corneal cells were infected with HSV-1. The direct immunofluorescent method was used to detect the presence of the herpes simplex antigen in HSV-1 infected corneal epithelial cells, keratocytes, and endothelial cells in vitro with fluorescein-labeled antiserum to HSV-1. Immunofluorescent localization of the HSV-1 antigen was expressed in all three types of corneal cells.
Animals ; Antigens, Viral/*analysis ; Cells, Cultured ; Cornea/*microbiology ; Herpesviridae Infections/*microbiology ; Rabbits ; Simplexvirus/*immunology

Adenoviridae ; immunology ; Antigens, Viral ; analysis ; Child ; Fluorescent Antibody Technique ; Humans ; Respiratory Syncytial Viruses ; immunology ; Respiratory Tract Infections ; immunology ; Respirovirus ; immunology ; Virus Diseases ; immunology

BACKGROUNDSeveral kinds of intercellular adhesion molecules closely relate to hepatitis B. The complex of CD(2) and CD58 plays an important role in enhancing the adhesion of T lymphocytes to target cells, hyperplasia and activation of T lymphocytes. In this study, we explored the relationship between the expression of CD58 in liver tissue and chronic hepatitis B infection.

METHODSWe determined the expression of the CD58 molecule on the surface of hepatocytes by using immunohistochemistry and the levels of serum HBV DNA from patients with HBV infection and from normal controls. The biochemical parameters of hepatic function were analyzed as well.

RESULTSCD58 expression in hepatocytes significantly increased with the severity progression of chronic HBV infection. The IOD levels (log10) of CD58 in the control, mild, moderate, and severe chronic HBV infection groups were 0, (7.20+/-4.64) x 10(3), (25.63+/-7.41) x 10(3) and (37.47+/-11.17) x 10(3) respectively (P<0.05 compared with the control group, respectively).

CONCLUSIONCD58 probably increases cell mediated immunity to eliminate hepatitis B virus and leads to damage of hepatocytes.

CD58 Antigens ; analysis ; DNA, Viral ; blood ; Hepatitis B, Chronic ; immunology ; Humans ; Immunohistochemistry ; Liver ; immunology ; physiopathology ; T-Lymphocytes ; physiology

BACKGROUNDTo study the composition and significance of the inclusion bodies of human cytomegalovirus (HCMV).

METHODSMicrodissection of inclusion bodies, PCR and Southern blot were adopted to detect DNA, and immunohistochemistry method and catalyzed signal amplification (CSA) were used to detect the different antigens of HCMV.

RESULTSThe inclusion bodies of HCMV were separated from the tissue section of human salivary gland. The fragments amplified by PCR from these dissected inclusion bodies were confirmed to be the DNA of HCMV. With the immunohistochemical method CSA, the immediately early and early antigens of HCMV were detected with monoclonal antibodies DDG9/CCH2, while matrix protein AAC10 was negative in the inclusion bodies.

CONCLUSIONThe ingredient of inclusion bodies of HCMV included the DNA and the antigens expressed in specific stage of the virus.

Antigens, Viral ; analysis ; immunology ; Cytomegalovirus ; genetics ; immunology ; Cytomegalovirus Infections ; diagnosis ; immunology ; virology ; DNA, Viral ; analysis ; genetics ; Humans ; Immunohistochemistry ; Inclusion Bodies ; chemistry ; immunology ; virology ; Microdissection ; Salivary Glands ; chemistry ; immunology ; virology

OBJECTIVETo develop the monoclonal antibody against N protein of SARS virus and study its applicability.

METHODSBALB/c mice were immunized with recombinant N protein. Spleen cells were collected and infused with SP2/0 cell. The infused cells were screened for anti-N protein antibody with ELISA. The positive cells were cloned and injected into abdominal cavity. The antibodies were purified from ascites. The affinities of those purified antibodies were analyzed with ELISA. The ELISA for detection of SARS virus antigen was developed by using antibody with the highest affinity. Its sensitivity and specificity were also evaluated primarily.

RESULTSEleven monoclonal cells secreting antibody have been developed. Three of the 11 purified monoclonal antibodies had very high affinity to N protein, while 4 purified McAbs showed very weak reaction to N protein, the affinities of remaining 4 McAbs were in between. The ELISA for detection of SARS virus antigen was developed with McAb 7. Its sensitivity was about 31 PFU/ml and had no cross reaction with other respiratory viruses.

CONCLUSIONThe monoclonal antibody has good specificity and may be used to detect SARS virus antigen. However, its sensitivity is to be evaluated further with clinical samples from SARS patients, especially at acute phase.

Animals ; Antibodies, Monoclonal ; biosynthesis ; Antibodies, Viral ; biosynthesis ; Antigens, Viral ; analysis ; Humans ; Hybridomas ; Mice ; Mice, Inbred BALB C ; Nucleocapsid Proteins ; immunology ; SARS Virus ; immunology ; Sensitivity and Specificity

OBJECTIVETo better understand the duplication of hepatitis B virus (HBV) in order to improve clinical diagnoses and treatments via quantitative measurement of HBV-DNA and comparison of correlation of HBV-DNA with HBeAg and anti-HBe.

METHODSFor 883 hepatitis B patients with positive HBsAg, HBV-DNA was measured by COBAS AMPLICOR HBV MONITOR reagent and COBAS AMPLICOR quantitative PCR instrument. Microparticle enzyme immunoassay analysis (MEIA) was then carried out with fully automatic enzyme immunoassay analysis instrument made by Abbott Axsym from the U.S. to measure HBeAg and anti-HBe. Correlation was analysed by SPSS.

RESULTS(1)Positive correlation between 690 HBV-DNA positive and HBeAg positive with r= 0. 505 (P< 0.01) was found with mean values as:HBV-DNA:7.12 x 10(12) copies/ml;HBeAg:218.31 S/CO. HBV-DNA:10(4) copies/ml, HBeAg: 104 S/CO; HBV-DNA: 10(5)-10(8) copies/ml, HBeAg: 112 S/CO; HBV-DNA: 10(9)-10(15) copies/ml, HBeAg: 252 S/CO. (2) No correlation was found between 193 HBV-DNA and anti-HBe + with r= -0.052(P= 0.477> 0.05) with Mean: HBV-DNA: 8.0x 10(10) copies/ml anti-HBe: 0.18 S/CO.

CONCLUSIONHBV-DNA and HBeAg appeared to have had linear correlation, showing that HBeAg> 100 S/CO,HBV-DNA> 10(4) copies/ml and hepatitis B virus were reproduced. However, HBV-DNA did not show linear correlation with anti-HBe as HBeAg negative and anti-HBe positive, the level of hepatitis B viral replication decrease slightly. But the virus load is still high. Infectivity can not neglect.

Antibodies, Viral ; analysis ; Carrier State ; DNA, Viral ; analysis ; Hepatitis B ; diagnosis ; genetics ; immunology ; Hepatitis B e Antigens ; analysis ; Hepatitis B virus ; genetics ; immunology ; Humans ; Immunoenzyme Techniques ; Polymerase Chain Reaction ; Viral Load ; Virus Replication

OBJECTIVETo establish a quantitative assay for enterovirus 71, this can be used in detecting the virus content during vaccine development and production.

METHODSWe established the method of quantitative assay for EV71 by using double antibody sandwich ELISA. The sensitivity, accuracy,precision and specificity of the method were evaluated.

RESULTSWe developed an ELISA method to quantitative assay for EV71. The quantitation limit of the method is 0.23 microg/ml and the quantitation scope of the method is 7.32-0.23 microg/ml, the coefficient correlation is R2 = 0.9976; The method showed good accuracy, precision and specificity. The recovery is between 90%-110% and the variation coefficient is lower than 10%.

CONCLUSIONAn ELISA method was developed for the quantitative assay of EV71 virus, which can be used for the rapid quantitative determination of EV71 virus during vaccine development and production.

Animals ; Antigens, Viral ; analysis ; immunology ; Cell Line ; Enterovirus ; immunology ; isolation & purification ; Enterovirus Infections ; diagnosis ; immunology ; virology ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Sensitivity and Specificity

The purpose of this study is to delineate the histopathologic findings of the spleen after Hantaan viral inoculation, which is the largest lymphoid organ in rats, and to identify the viral location by anti-Hantaan virus (HTNV) monoclonal antibody. All the sixty one suckling rats of less than twenty four hours of age were used. Except twenty one rats of control group, twenty-five rats inoculated intracerebrally for the early change and fifteen suckling rats inoculated intramuscularly for the late change were uniformly susceptible to lethal infection with the ROK 84-105-1 strain of seed HTNV. The characteristic histopathologic findings were; appearance of macrophages below the splenic capsule on the 3rd day, small lymphocytes around the periarteriolar sheath on the 5th day increasing in numbers on the 7th day, and a markedly expanded marginal zone with some immunoblasts and plasma cells as well as decreased extramedullary hematopoiesis on the 9th and 14th days. Time of onset of histopathologic changes in spleen thickness, appearance of medium and large lymphocytes and degree of extramedullary hematopoiesis were influenced by inoculation route, whereas expansion of the marginal zone was affected by postnatal age.
Animals ; Animals, Suckling ; Antigens, Viral/analysis ; Hantavirus/immunology ; Hematopoiesis ; Hemorrhagic Fever with Renal Syndrome/*pathology ; Rats ; Rats, Inbred Strains ; Spleen/*pathology

OBJECTIVEGene sequence data were clustered to explore evolution lineages of H3 antigen of influenza A virus.

METHODSAll data of H3 RNA sequence in NCBI Genbank and Influenza sequence database were downloaded and aligned in ClustalX while two step cluster method were applied to explore the data.

RESULTSAll sequences were aggregated into ten clusters, while seven of them mainly were human virus. Human virus and avian/other mammal virus were separated into different clusters distinctively, but coexisted into same clusters with swine virus. Time and host distribution were very distinctive in these clusters, but no geographic distribution features were found.

CONCLUSIONWith the interaction of human immunity system, H3 antigen mutated significantly every 5 - 7 years, and the speed of mutation had accelerated with the application of influenza vaccines in recent years. Mean while, human and swine influenza virus were not separated distinctly between clusters indicating that they had short inheritance distance. Result showed again that swine served as the mixer for antigenic recombination of different influenza virus.

Antigenic Variation ; genetics ; Antigens, Viral ; genetics ; Cluster Analysis ; Evolution, Molecular ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Hemagglutinins, Viral ; genetics ; immunology ; Humans ; Influenza A virus ; genetics ; immunology ; Mutation ; Sequence Analysis, DNA ; Viral Envelope Proteins ; genetics