Correlation analysis between serum NRSN2 with epstein-Barr virus antibody and DNA in nasopharyngeal carcinoma / 中国医师杂志

Objective@#To investigate the expression of serum Neurensin 2 (NRSN2) in patients with nasopharyngeal carcinoma (NPC), and to analyze its relationships with epstein-barr virus (EBV) antibody and EBV-DNA.@*Methods@#120 patients with NPC admitted to our hospital from April 2016 to May 2018 were selected as the study group, and 56 healthy people in the physical examination center were selected as the control group. Enzyme linked immunosorbent assay (ELISA) was used to detect the expressions of serum viral capsid antigen-IgA (VCA-IgA), nuclear related tumor antigen-IgA (EBNA-IgA) and early antigen-IgG (EA-IgG). The expression of serum NRSN2 and EBV-DNA were detected by fluorescence quantitative polymerase chain reaction (qPCR). Receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of serum NRSN2 level for NPC. Spearman was used to analyze the relationship between serum NRSN2 and EBV antibody with EBV-DNA in nasopharyngeal carcinoma, multivariate logistic regression analysis was used to analyze the risk factors of NPC.@*Results@#The positive rates of serum VCA-IgA, EBNA-IgA and EA-IgG in the study group were significantly higher than those in control group (P<0.05); the level of serum NRSN2 and the positive expression rate of EBV-DNA in the study group was significantly higher than that in the control group (P<0.05); ROC curves showed that the area under curve (AUC) of serum NRSN2 level in diagnosing NPC disease was 0.759, with sensitivity 63.33%, specificity 80.36%; there were significant positive correlations between serum NRSN2 with VCA-IgA, EBNA-IgA, EA-IgG and EBV-DNA in NPC patients (P<0.05); multivariate regression analysis showed that VCA-IgA, EBNA-IgA, EA-IgG, EBV-DNA and NRSN2 expressions were supportive predictive value for diagnostic NPC (P<0.05).@*Conclusions@#Up-regulation of serum NRSN2 levels in NPC patients is positively correlated with serum VCA-IgA, EBNA-IgA, EA-IgG and EBV-DNA levels, which may provide a reference for the prediction, diagnosis and treatment of NPC.

Correlation analysis between serum NRSN2 with epstein-Barr virus antibody and DNA in nasopharyngeal carcinoma / 中国医师杂志

Objective To investigate the expression of serum Neurensin 2 (NRSN2) in patients with nasopharyngeal carcinoma (NPC),and to analyze its relationships with epstein-barr virus (EBV) antibody and EBV-DNA.Methods 120 patients with NPC admitted to our hospital from April 2016 to May 2018 were selected as the study group,and 56 healthy people in the physical examination center were selected as the control group.Enzyme linked immunosorbent assay (ELISA) was used to detect the expressions of serum viral capsid antigen-IgA (VCA-IgA),nuclear related tumor antigen-IgA (EBNA-IgA) and early antigen-IgG (EA-IgG).The expression of serum NRSN2 and EBV-DNA were detected by fluorescence quantitative polymerase chain reaction (qPCR).Receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of serum NRSN2 level for NPC.Spearman was used to analyze the relationship between serum NRSN2 and EBV antibody with EBV-DNA in nasopharyngeal carcinoma,multivariate logistic regression analysis was used to analyze the risk factors of NPC.Results The positive rates of serum VCA-IgA,EBNA-IgA and EA-IgG in the study group were significantly higher than those in control group (P ＜0.05);the level of serum NRSN2 and the positive expression rate of EBV-DNA in the study group was significantly higher than that in the control group (P ＜ 0.05);ROC curves showed that the area under curve (AUC) of serum NRSN2 level in diagnosing NPC disease was 0.759,with sensitivity 63.33％,specificity 80.36％;there were significant positive correlations between serum NRSN2 with VCA-IgA,EBNA-IgA,EA-IgG and EBV-DNA in NPC patients (P ＜ 0.05);multivariate regression analysis showed that VCA-IgA,EBNA-IgA,EA-IgG,EBV-DNA and NRSN2 expressions were supportive predictive value for diagnostic NPC (P ＜ 0.05).Conclusions Up-regulation of serum NRSN2 levels in NPC patients is positively correlated with serum VCA-IgA,EBNA-IgA,EA-IgG and EBV-DNA levels,which may provide a reference for the prediction,diagnosis and treatment of NPC.

Genetic variations in open reading frame 2 gene of porcine circovirus type 2 isolated in Korea during 2016–2017 / 대한수의학회지

The capsid protein of porcine circovirus type 2 (PCV2) encoded by open reading frame 2 (ORF2) is important for neutralizing activity against PCV2 infection. This study investigated the heterogeneity of the ORF2 gene of PCV2 isolated in Korea during 2016–2017. The results revealed that PCV2d is currently the predominant genotype. Moreover, comparison of ORF2 from 17 PCV2 isolates revealed 88.3–100% homology at the nucleotide (deduced amino acid 86.3–100%) level. Interestingly, 61.5% (8/13) of the PCV2d isolates had glycine at position 210. These data provide a useful information for PCV2 epidemiology in Korea.

L1 Recombinant Proteins of HPV Tested for Antibody Forming Using Sera of HPV Quadrivalent Vaccine

Virus-like particles (VLPs) derived from human papillomavirus (HPV) L1 capsid proteins were used for HPV quadrivalent recombinant vaccine. The HPV quadrivalent vaccine is administrated in a 3-dose regimen of initial injection followed by subsequent doses at 2 and 6 months to prevent cervical cancer, vulvar, and vaginal cancers. The type 6, 11, 16, or 18 of HPV infection is associated with precancerous lesions and genital warts in adolescents and young women. The HPV vaccine is composed of viral L1 capsid proteins are produced in eukaryotic expression systems and purified in the form of VLPs. Four different the L1 protein of 3 different subtypes of HPV: HPV11, HPV16, and HPV18 were expressed in Escherichia coli divided into 2 fragments as N- and C-terminal of each protein in order to examine the efficacy of HPV vaccine. Vaccinated sera failed to recognize N-terminal L1 HPV type 16 and type 18 by western blot while they detected N-terminal L1 protein of HPV type 11. Moreover, the recombinant C-terminal L1 proteins of type 16 was non-specifically recognized by the secondary antibody conjugated with horseradish peroxidase. This expression and purification system may provide simple method to obtain robust recombinant L1 protein of HPV subtypes to improve biochemical analysis of antigens with immunized sera.

Inhibitory effects of the Chlamydiaphage phiCPG1 capsid protein Vp1 on Chlamydia psittaci strain guinea pig inclusion conjunctivitis and Chlamydia trachomatis serovar E / 中华皮肤科杂志

Objective To evaluate inhibitory effects of the Chlamydiaphage phiCPG1 capsid protein Vp1 on Chlamydia psittaci strain guinea pig inclusion conjunctivitis (GPIC) and Chlamydia trachomatis (Ct) serovar E,and to provide new ideas for the treatment of Ct infection.Methods The Chlamydiaphage phiCPG1 capsid protein Vp1 was expressed in Escherichia coli BL21 transfected with the recombinant plasmid Vp1-pET30a (+),identified by Western blot analysis and purified by using dialysis bags.Bicinchonininc acid (BCA) assay was performed to determine the concentration of Vp1 protein.GPIC and Ct serovar E strains were both classified into 4 groups to be firstly incubated with Vp1 protein (Vp1 group),Tris-glycine solution (Tris group),S protein (S group) or Dulbecco's Modified Eagle Medium (DMEM,DMEM group) at room temperature for 3 hours,then were used to infect Hela cells followed by 72-hour (GPIC) or 48-hour (Ct serovar E) culture with the presence of Vp 1 protein (Vp 1 group),Tris-glycine solution (Tris group),S protein (S group) or DMEM (DMEM group).Subsequently,immunofluorescence staining was conducted to observe and count chlamydial inclusions.Results The number of GPIC inclusions was significantly different between the 4 groups after 72-hour culture (F=476.632,P＜ 0.05),and lower in the Vp1 group (5.0 ± 1.5) than in the Tris group (24 ± 1.2,P＜ 0.05),S group (25 ± 1.7,P＜ 0.05) and DMEM group (25 ± 1.5,P＜ 0.05),but insignificantly different between the latter 3 groups (P ＞ 0.05).Compared with the DMEM group,the Vp1 group showed a significant decrease of 80.2％ ± 3.99％ and 77.2％ ± 1.79％ in the number of GPIC and Ct serovar E inclusions respectively,with no significant difference in the inhibitory effect of Vp1 on GPIC versus Ct serovar E (t =2.057,P ＞ 0.05).Conclusion The phiCPG1 capsid protein Vp1 can obviously inhibit GPIC and Ct serovar E infections to a similar degree.

VCA-IgA and Rta-IgG joint detection diagnosis and effectiveness of nasopharyngeal carcinoma / 中华检验医学杂志

Objective To investigate the clinical efficacy of combined detection of VCA-IgA and Rta-IgG in the diagnosis of nasopharyngeal carcinoma.Methods From May 2013 to November 2014, 3 913 serum samples(male 2 367,female 1 546) from healthy people who had health examination in our medical center were collected and 169 serum samples(male 118,female 51) were collected from the patients who were diagnosed as nasopharyngeal carcinoma by pathological biopsy.Serum samples in two groups were detected by EBV RTA-IgG, VCA-IgA assay ( ELISA ) respectively.SPSS17.0 statistical software and receiver operating characteristic curve ( ROC) were applied to data analysis.Results The Rta-IgG positive rates of EB virus were 93.5%in NPC group (158/169) and 2.4%(93/3 913) in healthy group;while the VCA-IgA positive rates were 79.3%in NPC group ( 134/169 ) and 8.9% ( 349/3 913 ) in healthy group. The sensitivity(χ2 =14.49,P<0.05) and specificity(χ2 =157.15,P<0.05) of Rta-IgG in the diagnosis of nasopharyngeal carcinoma were significantly better than that of VCA-IgA. Using VCA-IgA/Rta-IgG combined detection analysis, not only failed to effectively improve the diagnosis of nasopharyngeal cancer, but to reduce the detection sensitivity to 72.8%( 123/169 ) , compared with Rta-IgG detection only. Conclusions Rta-IgG is significantly better than that of VCA-IgA.There was no significant improvement in the clinical diagnostic efficacy of nasopharyngeal carcinoma using VCA-IgA/Rta-IgG combined detection mode.

Bioinformatic analysis of non-VP1 capsid protein of coxsackievirus A6 / 华中科技大学学报（医学）（英德文版）

This study bioinformatically analyzed the non-VP1 capsid proteins (VP2-VP4) of Coxasckievirus A6 (CVA6), with an attempt to predict their basic physicochemical properties, structural/functional features and linear B cell eiptopes. The online tools SubLoc, TargetP and the others from ExPASy Bioinformatics Resource Portal, and SWISS-MODEL (an online protein structure modeling server), were utilized to analyze the amino acid (AA) sequences of VP2-VP4 proteins of CVA6. Our results showed that the VP proteins of CVA6 were all of hydrophilic nature, contained phosphorylation and glycosylation sites and harbored no signal peptide sequences and acetylation sites. Except VP3, the other proteins did not have transmembrane helix structure and nuclear localization signal sequences. Random coils were the major conformation of the secondary structure of the capsid proteins. Analysis of the linear B cell epitopes by employing Bepipred showed that the average antigenic indices (AI) of individual VP proteins were all greater than 0 and the average AI of VP4 was substantially higher than that of VP2 and VP3. The VP proteins all contained a number of potential B cell epitopes and some eiptopes were located at the internal side of the viral capsid or were buried. We successfully predicted the fundamental physicochemical properties, structural/functional features and the linear B cell eiptopes and found that different VP proteins share some common features and each has its unique attributes. These findings will help us understand the pathogenicity of CVA6 and develop related vaccines and immunodiagnostic reagents.

The natural history of cervical intraepithelial neoplasia Ⅰ and the clinical significance of p16INK4a protein as a marker of progression in cervical intraepithelial neoplasiaⅠ / 中华妇产科杂志

Objective To describe the natural history of cervical intraepithelial neoplasia(CIN)Ⅰand the biologic factors associated with the progression of CINⅠ and to analyze the predictive values of p16INK4a protein for the progression of CINⅠ. Methods From August 2010 to July 2013, 104 patients referred for abnormal cytology [≤ low-grade squamous intraepithelial lesion (LSIL); including negative for intraepithelial lesion or malignancy (NILM), atypical squamous cells of undetermined significance (ASCUS), LSIL] and high-risk (HR) HPV positive,and were diagnosed CINⅠ by colposcopy-assisted biopsy and followed at 1-year intervals in the First Affiliated Hospital of Nanjing Medical University. In order to analyze the relationship between the progression of CINⅠ with clinical biologic factors, including patient age, cervical cytology before colposcopy, loads of HR HPV, HPV L1 capsid protein, p16INK4a protein,χ2 tests was used to compare the different frequencies of factors in groups of progressed and persisted/regressed CINⅠ, then five factors with progressed CINⅠwere processed into binary logistic regression analysis. Results (1) In the first year of follow-up, among 104 patients(including 15 cases NILM,78 cases ASCUS,11 cases LSIL), 52 cases of them were NILM and HR HPV negative, 30 cases were negative for intraepithelial lesion, 10 cases were CINⅠ, 5 cases were CINⅡand 7 cases were CINⅢ. In total, 82 cases (78.8%,82/104) cases had regressed, 10 cases (9.6%,10/104) persisted, 12 cases (11.5%,12/104) progressed [including 5 cases (4.8%,5/104) progressed to CIN Ⅱ, 7 cases (6.7%,7/104) progressed to CIN Ⅲ, none progressed to invasive cancer]. (2) All patients, 88 cases of them accepted immunohistochemical detection the expression of p16INK4a protein. The result shown that 30 cases (34%,30/88) were positive and 58 cases (66%,58/88) were negative. And 94 cases accepted immunocytochemical detection the expression of HPV L1 capsid protein, 49 cases (52%,49/94) were positive and 45 cases (48%,45/94) were negative. (3) Univariate analysis showed that age of the patient, loads of HR HPV, cervical cytology before colposcopy and the expression of HPV L1 capsid protein were not risk factors of the progression of CINⅠ(all P>0.05) except for the expression of p16INK4a protein (P<0.05). Multivariable analysis found that p16INK4a protein positive was associated with progression of CINⅠ(OR=5.1,95%CI:1.162-22.387,P=0.031). (4) Thirty-one cases were p16INK4a protein positive, 8 cases (27%,8/30) of them progressed,while 4 cases (7%,4/58) of 58 cases with p16INK4a protein negative progressed,in which there were significant difference (P<0.05). The sensitivity was 75%, the specificity was 71%, the positive predictive value was 27%and the negative predictive value was 93%for progression to CINⅡ-Ⅲof p16INK4a protein staining. Conclusions The progression rate of CINⅠwith abnormal cytology (≤LSIL) and HR HPV positive was lower, and there was no progression to invasion at 1-year intervals. Immunostaining of p16INK4a protein as the risk factors of CINⅠprogression could have a role in prediction of CINⅠand the management of high-risk CINⅠ.

Clinical significance of HPV L1 capsid protein detection in cervical exfoliated cells in high-risk HPV positive women / 中华妇产科杂志

Objective To explore the clinical significance of human papillomavirus L1 capsid protein detection in cervical exfoliated cells in high-risk HPV positive women. Methods From November 2012 to June 2013,386 high-risk HPV positive (detected by hybrid capture Ⅱ) cases were enrolled as eligible women from Huzhou Maternity&Child Care Hospital and Women′s Hospital,School of Medicine, Zhejiang University. All eligible women underwent liquid-based cytology (ThinPrep) followed by colposcopy. Biopsies were taken if indicated. Cervical exfoliated cells were collected for HPV L1 capsid protein detection by immunocytochemistry. Expression of HPV L1 capsid protein in groups with different histological diagnosis were compared, and the role of HPV L1 capsid protein detection in cervical exfoliated cells in cervical lesions screening was accessed. Results Total 386 enrolled eligible women were finally diagnosed histologically as follwed:162 normal cervix, 94 low-grade squamous intraepithelial lesion (LSIL), 128 high-grade squamous intraepithelial lesion (HSIL) and 2 squamous cervical cancer (SCC). The positive expression rate of HPV L1 in HSIL+(HSIL or worse) group was significantly lower than that in LSIL-(LSIL or better) group (19.2% vs 66.4%,P=0.000). While identifying HSIL+ in HPV positive cases and compared with cytology, HPV L1 detection resulted in significant higher sensitivity (80.77%vs 50.77%,P=0.000) and negative predictive value (NPV;87.18% vs 76.47%,P=0.004), significant lower specificity (66.41% vs 81.25%,P=0.000),and comparable positive predictive value (PPV;54.97% vs 57.89%, P=0.619). To identify HSIL+in HPV-positive/cytology-negative women, the sensitivity, specificity, PPV, and NPV of HPV L1 detection were 87.50%, 61.54%, 41.18%, and 94.12%respectively, while 80.00%, 86.36%, 80.00%and 86.36%respectively in HPV-positive/atypical squamous cell of undetermined significance(ASCUS)women. Conclusions HPV L1 capsid detection in cervical exfoliated cells have a role in cervical lesions screening in high-risk HPV positive women, and may be a promising triage for high-risk HPV-positive/cytology-negative or ASCUS women.

Preliminary observation on antitumor effect of HPV58 composite DNA vaccine / 中华妇产科杂志

Objective To initially observe the antitumor immune of PVAX1-HPV58mE6E7FcGB composite DNA vaccine.Methods Before detecting immune effect of the vaccine,the B16-HPV58E6E7 tumor cell line was built which could steadily express HPV58E6E7 fusion gene.Then,HPV58E6E7-GST fusion protein as an antigen was expressed and purified.Before or after immunized with the vaccine,the C57BL/6 mice were challenged by B16-HPV58E6E7 cells.Anti-tumor transplantation and tumor growth inhibition experiment were performed to observe prevention and treatment effects on the vaccine.Specific humoral and cellular immune responses in the immunized mice were detected by ELISA,enzyme linked immunospot assay (ELISPOT) and cytotoxic T lymphocyte (CTL) method.Results In the anti-tumor transplantation experiment,tumor formation rate was only 9/15 in the mice which were immunized by PVAX1-HPV58mE6E7FcGB vaccine,time before tumor formation was the longest [(13.6 ± 1.7) days] and tumor growth was the slowest in the vaccine group.In tumor growth inhibition experiment,inhibition rate reached 81.4％ in the vaccine group.Except tumor formation rate,all data in the vaccine group was superior to the pure antigen PVAX1-HPV58mE6E7Fc group (P ＜ 0.05).Humoral immune effect showed that both the vaccine and the pure antigen could induce specific antibody in the immunized mice,and the highest titer were 1 ∶ 25600 and 1 ∶ 12800,respectively.Although there was not significant difference of antibody titer between the vaccine and the pure antigen group (P ＞ 0.05),the number of activated T cells in the vaccine group was almost four times as that in the pure antigen group [(219 ±34)/4 × 105 spleen lymphocytes versus (55 ±25)/4 × 105 spleen lymphocytes,P ＜ 0.05],and the highest specific CTL that vaccine induced was significantly higher than that of pure antigen (43.3％ versus 31.3％,P ＜ 0.05).Conclusions Humoral and cellular immune response could be effectively stimulated by PVAX1-HPV58mE6E7FcGB composite DNA vaccine.Growth of B16-HPV58E6E7 cells was significantly inhibited in the immunized mice.The cellular immune effect on the vaccine was superior to the pure antigen.Therefore,PVAX1-HPV58mE6E7FcGB could be used as a candidate vaccine for immune therapy to the HPV58 positive tumors and precancerous lesions.

The prevalence of human bocavirus among children with acute respiratory tract infection in Guangdong / 中华传染病杂志

Objective To investigate the prevalence of human bocavirus(HBoV)among children with acute respiratory tract infection(ARTI)in Guangdong Province.Methods Four hundred and forty-seven nasopharyngeal aspirates or swabs samples from children with ARTI in Guangdong Province were collected from June 2007 to May 2008.HBoV capsid protein VP gene fragments were detected using polymerase chain reaction(PCR).Positive PCR products were sequenced.The DNA and translated amino acid sequences were aligned with known HBoV sequences in GenBank and phylogenetic analysis was also done.Results The positive rate of HBOV was 5.1%of samples from 447 ARTI cases.Ten samples were positive for both HBoV and other respiratory virus,which was 43.5%of positive samples.The main diagnosis for HBoV positive children included wheezing pneumonia,bronchiolitis and bronchial pneumonia.HBoV positive children ranged from 42 days to 6 years old,and most of them were younger than one year.HBOV infection was more common during summer,early autumn and late spring.Through sequence alignment and phylogenetie analysis,the DNA sequences and amino acid sequences of VP gene fragments of isolated HBoV strains showed 97.8%-98.8%and 99.3%-100.0%identity with ST1,respectively.Conclusions HBoV is one of the important pathogens of lower respiratory tract infection in children in Guangdong Province,which is more prevalent in infants younger than one year.Although VP gene fragment of HBoV is conservative in general,there are still some missense mutations.

Preparation and preliminary application of monoclonal antibody against Vp1 protein of chlamydiaphage ΦCPG1 / 中华皮肤科杂志

Objective To express recombinant Vp1 protein of chlamydiaphage φCPG1, prepare monoclonal antibody against Vp1 protein and utilize it to screen clinical isolates of Chlamydia trachomatis. Methods The Vp1 protein was obtained by prokaryotic expression, and monoclonal antibody against this protein was prepared by hybridoma technique. ELISA and Western blot were used to identify monoclonal antibodies. Then,the monoclonal antibody was prepared in quantity by injecting hybridoma cells into the abdominal cavity of BALB/C mice, and purified by using protein G affinity chromatography. Clinical isolates of Chlamydia trachomatis were screened for the chlamydiaphage by immumofluorescence assay using the prepared monoclonal antibody.Results Purified Vp1 protein was obtained. The monoclonal antibody against Vp1 protein was gained after 3times of sub-clone and consistently identified as IgG1. Three hybridoma cell strains that stably secreted monoclonal antibody were generated. Chromosome analysis of hybridoma cells showed that the mean number of chromosome was 96, most of them were telocentric and a few were submetacentric. The titer of purified monoclonal antibody was more than 1: 12 800. Twenty clinical isolates were screened by using the monoclonal antibody and no positive results were obtained. Conclusions The monoclonal antibody against Vp1 protein of chlamydiaphage φCPG1 is successfully prepared, while no chlamydiaphage is detected by immumofluorescence assay using the prepared antibody in 20 clinical isolates of Ct.

Cloning, expression and identification of phage's capsid Vp3 protein of guinea pig inclusion conjunctivitis chlamydia / 中华传染病杂志

Objective To get phagc's capsid Vp3 gene and protein of guinea pig inclusion conjunctivitis (GPIC) chlamydia. Methods The genome DNA was extracted from the φCPG1 phage.The full sequence of Vp3 gene was amplified by polymerase chain reaction (PCR) from the above genome DNA. The Vp3 gene was digested by restriction endonuclease and then inserted into prokaryotic plasmid vector pET30a (+). The recombinant plasmid was transformed into E. coil BL21, and was identified by restriction endonuclease, PCR and sequencing. The E. coil BL21 with expected recombinant plasmid was induced and the expressed recombinant Vp3 protein was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, then purified by agarose gel. Results The recombinant gene was sequenced and proved to be 447 bp which was consistent with the φCPG1 Vp3 gene sequence in GenBank. A 25 000 capsid protein was expressed and confirmed by SDS-PAGE and Western blot. The purified protein was obtained. Conclusion The capsid Vp3 protein of φCPG1 is successfully expressed and purified, which is helpful for the further study on its mechanism and clinical applications.

Detection of human papillomavirus L1 capsid protein expression in liquid-based cytology samples with abnormal cytology / 中华妇产科杂志

Objective To investigate the possibility of detection of the human papillomavirus (HPV)L1 capsid protein to predict the coarse of mild or moderate cervical intraepithelial neoplasia(CIN).Methods Immunocytochemical analysis using antibody against HPV L1 capsid protein was carried out on 274 samples obtained from women performed Tri Path Pap tests.positive for high-risk HPV DNA detected by hybrid capture Ⅱ (HC-Ⅱ)or cytologic diagnosed atypical squamous cells of undetermined significance (ASCUS)or more severe.For cytological diagnosed,there were ASCUS 105 cases,low-grade squamous intraepithelial lesions (LSIL) 119 cases,atypical squamous cells cannot exclude high-grade squamous intraepithelial lesion (ASC-H)9 cases,high-grade squamous intraepithelial lesions(HSIL)36 cases,and squamous cell carcinoma(SCC)5 cases.But for the pathologic diagnosed,there were chronic cervicitis 96 cases.CIN Ⅰ 55 cases,CIN Ⅱ 55 cases,CIN Ⅲ 32 cases,and SCC 6 cases.Results Of the 274 cases,HPV L1 capsid protein was positive in 69.8%(67/96) of cervicitis,83.5%(71/85)of CIN Ⅰ,41.8% (23/55)of CIN Ⅱ,3.1%(1/32)of CIN Ⅲ and 0(0/6)of SCC.Cytologic diagnosis revealed a higher expression rate in LSIL(75.6%.90/119) than that in ASCUS(63.8%,67/105)or in HSIL + SCC (9.8%,4/41;all P<0.01).Of 71 cases with ASCUS and ISIL without treated,none of HPV L1 positive cases(0/55)progressed in cytology,while 19%(3/16)of HPV L1 negative cases progressed to ASC-H,HSIL(P<0.01).Conclusion The expression rates of HPV L1 protein in liquid-based cell specimen is decreased as the cytopathology diagnosis severe degree.which may imply the histopathology diagnosis of cervix,predict the progression of cervical lesion,and help to treat the cases with ASCUS and LSIL.

Detection of antibodies to Epstein-Barr virus with ELISA based on fusion capsid protein / 中华检验医学杂志

and 98.6% (436/442), respectively. Conclusion The ELISA based on fusion viral capsid proteins is sensitive, specific and accurate method for determining antibodies to VCA of EBV for both clinical diagnosis and epidemiology studies.

Prokaryotic expression and application of human herpes virus 8 small capsid protein encoded by open reading frame 65 / 中华检验医学杂志

Objective To construct prokaryotic vector and express human herpes virus 8 (HHV8) small capsid protein open reading frame 65 (ORF65) in Escherichia coli.Methods DNA was extracted from BCB-1 cells (HHV8-positive but EBV-negative).HHV80RF65 coding sequence was amplified by PCR and inserted into the prokaryotic expression vector pThioHisA.Recombinant plasmid was transformed into E.coli BL21 to express fusion protein induced by IPTG.The expressed products were purified by affinity chromatography on Ni-NTA resin.The antigenieity and the specificity of the recombinant proteins was identified by Western blot with HHVS-positive serum samples.ELISA coating with the recombinant proteins was used to screen 568 serum samples,which were simultaneously detected by IFA kit(Biotrin) and ELISA kit(Qiagen).Results Gene sequencing showed that the target gene was identical with that of HHV8 standard species,the 31 500 fusion protein was found in SDS-PAGE.ELISA coating with the recombinant proteins had a good agreement with IFA kit(Biotrin) and ELISA kit(Qiagen).The detection for the clinical samples showed the ELISA kit was feasible,applicable and consistant.Conclusion The recombinant proteins showed good antigenicity,and it is valuable for further study on HHV8 specific antibody detection.