Comparison of ELISA diagnostic kits used in China for hepatitis C virus IgG antibody detection / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To compare the commercial diagnostic reagent available in China for hepatitis C virus ( HCV) IgG antibody detection in order to provide some useful information for HCV prevention.</p><p><b>METHODS</b>The HCV-IgG detection reagents produced by six different Enterprises named A, B, C, D, E and F were chosen and applied to detect 160 HCV specious sera samples. HCV-IgG reagent from ABBOTT was adopted as gold-standard and the samples in gray zone were determined by RIBA method finally.</p><p><b>RESULTS</b>160 sera samples comprised 88 positive samples and 72 negative samples. The total conformity ranged from 88.13% to 98.13% and the Youden indexes ranged from 0.74 to 0.96 when the reagents from six different Enterprises were compared with gold-standard. The highest conformity was 98.13%, observed in D reagent with the highest Youden index of 0.96.</p><p><b>CONCLUSION</b>The total conformity rates were more than 88% when the HCV-IgG antibody detection reagents from six different Enterprises were compared with ABBOTT reagent. It was highly conformable. However, some reagent proved to be less conformable in negative samples detection.</p>

Hepatitis C viruses infection situation in the human population of six provinces in China / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the seroprevalence of hepatitis C viruse infection in the human population in six regions of Beijing, Heilongjiang, Shandong, Ningxia, Gansu and Sichuan in China.</p><p><b>METHODS</b>ELISA was used for detecting anti-HCV IgG of the serum samples. All sample were collected in 2006-2008 in six areas.</p><p><b>RESULTS</b>9538 samples were detected. The total positive rate of anti-HCV was 0.39% (37), 0.23% (3/1328) in Beijing, 0.74% (12/1629) in Heilongjiang, 0.26% (5/1962) in Shandong, 0.1% (1/1000) in Ningxia, 0.44% in Gansu (9/2 037) and Sichuan (7/1582), respectively. The 37 positive samples at the sex were 19 (51.35%) of man and 18 (48.65%) of female. At the age group were 1 (2.70%) of < 10 years old, 5 (13.51%) of 10-19 years old, 4 (10.81%) of 20-29 years old, 6 (16.22% ) of 30-39 years old, 9 (24.32%) of 4049 years old, 12 (32.43%) of > or = 50 years old. The positive samples were detected anti-HAV-IgG, anti-HEV-IgG and HBsAg/ HBcAb /HBeAb. The positive Number was 35 (94.59%), 10 (27.03%) and 2 (5.41%) respectively.</p><p><b>CONCLUSIONS</b>HCV infection rate in the population was 0.1% -0.74%, 1/3 of > or = 50 years old in HCV positive. Hepatitis C virus co-infection with HAV, HEV and HBV.</p>

Hepatitis viruses infection situation in human population of the Gansu province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the seroprevalence of hepatitis viruses in human population of Ganso province.</p><p><b>METHODS</b>ELISA was used for detecting anti-HAY IgG, HBsAg/HBsAb, anti-HCV IgG and anti-HEV IgG of the serum samples. All sample were collected in four areas of KL, LT, HN and ZhL of Gansu province in 2008.</p><p><b>RESULTS</b>1977 samples were detected, The positive rates of anti-HAY, HBsAg/HBsAh, anti-HCV, and anti-HEY are 84.57% (1672/1977), 4.81% (95/1977) and 28.73% (568/1977), 0.46% (9/1977),and 13.10% (259/1977), respectively. The positive rate at different age group, for anti-HAY was 43.21% of c 9 years old, 80.23% of 10-19 years old, 93.02% of 20-29 years old, 95.55% of 30-39 years old, 95.60%-97.52% of 40-60 years old. For HBsAg/HBsAb were 2.09% or 29.27%, 3.02% or 21.81%, 6.20% or 36.43%, 5.93% or 31.16%, 6.43% or 3l.43%, 7.32% or 24.88%, 3.33% or 28.89% at the same age group, respectively, for anti-HCV, was 0.59% of 30-39 years old, 1.79% of 40-49 years old, 0.98% of 50-59 years old. For HEY-IgG was 3.48% of < or =9 years old, 6.05% of 10-19 years old, 8.53% of 20-29 years old, 19.29%-21.67% of 30- > or =60 years old.</p><p><b>CONCLUSIONS</b>The inoculation againt HAY and HBY is enhanced in the youngster population. HBsAg carrier and UCY infection is decreasing. The HEY infection in the Hid people is lower than the Han nationality.</p>

Antibody detection of hepatitis E virus in some human population, swine and chicken in Beijing in China / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the seroprevalence of HEV infection in human population, swine and chicken in Beijing region.</p><p><b>METHODS</b>EIA was used for detecting anti-HEV IgG of the serum samples. All samples were collected in 2006-2007 in Beijing areas.</p><p><b>RESULTS</b>The anti-HEV IgG was detected positive in 21.52% of human (260/1208), 46.88 % (15/32) of swine, but was negative in chickens (0/24). The positive rate of human at different age group, was 5.60% (14/250) of 11-20 year, 20% (42/210) of 21-30 year, 24.03% (62/258) of 31-40 year, 26.44% (78/295) of 41-50 year, 32.82% (64/195) of 51-60 year. The male (29.51%) was higher than the female (21.70%).</p><p><b>CONCLUSION</b>The HEV infection was correlation with age and sex significantly. The infection rate was increased with age, the positive rate in swine was more double than the human population.</p>

Hepatitis viruses infection situation in Mianyang of the Sichuan province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the seroprevalence of hepatitis viruses in Mianyang of the Sichuan province.</p><p><b>METHODS</b>EIISA was used for detecting anti-HAV IgG, HBsAg/HBsAb, anti-HCV IgG and anti-HEV IgG of the serum samples. All sample were collected in Mianyang areas in 2007.</p><p><b>RESULTS</b>1352 samples were detected. The positive rates of anti-HAV, HBsAg/HBsAb, anti-HCV,and anti-HEV are 81.07% (1096/1352), 5.40% (73/1352) and 61.32% (829/1352), 0.37% (5/1352) and 49.26% (666/1352), respectively. The positive rate at different age group, for anti-HAV was 38.21% of 10-19 years old, 83% of 20-29 years old, 88% of 30-39 years old, 95.03% of 40-49 years old, 97% of 50-59 years old, 97.77% of 60-69 years old, 97.52% of > or =70 years old. For HBsAg/HBsAb were 5.65% or 50.83%, 10.0% or 68.0%, 5.20% or 78.80%, 5.97% or 78.11%, 6.50% or 62.50%, 1.12% or 51.40%, 4.96% or 30.58% at the same age group, respectively,for anti-HCV, was 0.33% of 10-19 years old, 0.80% of 30-39 years, 0.56% of 60-69 years old, 0.83% of > or =70 years old.For HEV-IgG was 26.58% of 10-19 years old, 42.0% of 20-29 years old, 55.22%-61.0% of 30-> or =70 years old, for anti-HEV IgM, was 10.06% (53/527) in the positive samples of HEV-IgG.</p><p><b>CONCLUSION</b>The inoculation againt HAV and HBV is enhanced in the young population. HBsAg carrier and HCV infection is decreasing. The HEV infection is actually increasing.</p>

Antibody detection and sequencing analysis of hepatitis E virus in human population, swine and chicken in Sichuan region in China / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the seroprevalence of HEV infection and genotype.</p><p><b>METHODS</b>ELISA were used for detecting anti-HEV IgG of the serum samples, the nested reverse transcriptase PCR (RT-nPCR) was used for detecting HEV RNA in patient serum and swine bile samples. All samples were collected in 2005-2007 in some districts in Sichuan province. The primers used for genotyping were the ORF2 region of HEV genome.</p><p><b>RESULTS</b>The anti-HEV IgG was detected positive in childrens 6.10% (41/672), adults 42.26% (280/ 661), swines 88.89% (32/36), chickens negative (0/59). 1 case of 15 serum samples of anti-HEV IgM positive and 3 of 54 swine bile samples were positive for HEV RNA by RT-PCR.Sequence analysis of 4 isolates has 92.1% to 98.6% nucleotide sequence homology. These isolates from human and swine were identified closely related to Ch-T21 strain 90.1%-96.9% sequence homology, which belonged to HEV genotype 4B.</p><p><b>CONCLUSIONS</b>The swine were the risk factors in the spread of hepatitis E virus.</p>

Evaluation of the ELISA diagnostic kits for hepatitis E virus antibody in the reference serum, the suspect patients of hepatitis E and normal persons' sera / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To evaluate the quality of the ELISA diagnostic kits for detecting hepatitis E virus (HEV) specific IgG antibody.</p><p><b>METHODS</b>Five diagnostic kits (WT, GD, HM, GeneLabs and KH) for anti-HEV IgG were assayed by detecting HEV IgG in the HEV diagnostic reference sera from 24 positive cases and 30 negative cases. 42 cases clinical suspect patients with hepatitis E from Ditan hospital in Beijing in 1994 to 1995 and 230 normal persons' sera from Chengdu city, Sichuan province in September in 2005.</p><p><b>RESULTS</b>The conformity rates of positive and negative sera was 95.83% (23) and 96.67% (29) for WT kit, 87.50% (21) and 100% (30) for GD kit, 87.50% (21) and 96.67% (29) for HM kit, 66.67% (16) and 100% (30) for GeneLabs kit, 45.83% (11) and 100% (30) for KH kit for detecting anti-HEV positive and negative reference sera, respectively. The five kits were used to detect anti-HEV IgG among 42 clinical suspect patients with hepatitis E and 230 normal persons' sera, The positive rate was 97.62% (41) and 31.74% (73) by WT, 100% (42) and 17.39% (40) by GD kit, 97.62% (41) and 23.91% (55) by HM kit, 90.48% (38) and 7.83% (18) by GeneLabs kit, 90.48% (38) and 3.48% (8) by KH, respectively.</p><p><b>CONCLUSION</b>The positive rates of all the 5 kits were over 90% and had a very high conformity in detecting anti-HEV IgG in clinical patient with hepatitis E, positive rates of 3.48% to 31.74% were found in detecting the normal persons' sera. The research showed insufficiency of the ELISA kits for epidemiological survey of HEV infection in population.</p>

Expression of the hepatitis Delta antigen in prokaryotic cell and evaluation of its application as an EIA diagnostic reagent / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To construct the pRSETB-HDAg recombinant expression plasmid and to obtain soluble hepatitis D virus antigen with high biological and antigenic activity.</p><p><b>METHODS</b>HDAg gene fragment was inserted into fusion expression pRSET B vector that includes T7 promoter and a polyhistidine tag. The recombinant plasmid was transformed into host bacterium BL21 after induction with IPTG. The expression supernatant was purified by chelating affinity chromatography and the recombinant HDAg antigenic activity was detected by EIA.</p><p><b>RESULTS</b>EIA detection using the recombinant HDAg showed strong positive reaction with hepatitis D patients sera. The positive rates of the EIA, compared with HDAg from USA and Hua Mei EIA kit in detecting 26 cases of anti-HDV positive reference sera, were 100%, 96.15% and 100%, respectively.</p><p><b>CONCLUSION</b>Recombinant plasmid for HDAg with good antigenicity was successfully constructed and could be used as hepatitis D antibody detection reagent.</p>

Antigencity identification of recombinant hepatitis E virus ORF2 protein expressed in Pichia pastoris / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To determine the antigenicity of recombinant hepatitis E virus ORF2 (rHEV ORF2) protein expressed in Pichia pastoris (P. pastoris).</p><p><b>METHODS</b>By using the rHEV ORF2 protein from E.coli as control, an indirect ELISA was adopted to identify the sensitivity, specificity and stability of rHEV ORF2 protein from P. pastoris in detection of HEV IgM and IgG antibody in sera from patients with hepatitis E. The reactivity of the rHEV ORF2 against 5 HEV ORF2 monoclonal antibodies (McAbs) was also tested.</p><p><b>RESULTS</b>The minimum concentration of coated antigen with which HEV IgG could be detected was 12.5 ng/ml, while the highest serum dilution to detect both IgM and IgG antibodies against HEV was 1:5 120. No cross-reaction was found with sera from patients with any other types of hepatitis. The 37 degree C acceleration test showed that the rORF2 was highly stable within 12 months at 4 degrees C. The 5 HEV ORF2 McAbs showed better reaction with the rORF2 from P. pastoris, especially that 4B2, 2E2, whose reaction against the rORF2 were 125 and 25 times respectively higher than that of rORF2 from E.Coli.</p><p><b>CONCLUSION</b>There may be more extensive conformational epitopes in the rHEV ORF2 from P. pastoris. The excellent antigenicity, sensitivity and stability suggest that it can be served as a new candidate antigen for the development of diagnostic reagents of hepatitis E.</p>