Clinical Characteristics of Children with Hemophagocytic Syndrome with Different EB Virus DNA Loads / 中国实验血液学杂志

OBJECTIVE@#To analyze the clinical characteristics of hemophagocytic syndrome (HLH) children with different EB virus (EBV) DNA loads, and to explore the relationship between differential indicators and prognosis.@*METHODS@#Clinical data of 73 children with HLH treated in our hospital from January 2015 to April 2022 were collected. According to EBV DNA loads, the children were divided into negative group (≤5×102 copies/ml), low load group (>5×102-<5×105 copies/ml) and high load group (≥5×105copies/ml）. The clinical symptoms and laboratory indexes of the three groups were compared, and the ROC curve was used to determine the best cut-off value of the different indexes. Cox regression model was used to analyze the independent risk factors affecting the prognosis of children, and to analyze the survival of children in each group.@*RESULTS@#The proportion of female children, the swelling rate of liver and spleen lymph nodes and the involvement rate of blood, liver, circulation and central nervous system in the high load group were higher than those in the negative group. The incidence of disseminated intravascular coagulation(DIC) and central nervous system(CNS) involvement in the high load group were higher than those in the low load group. The liver swelling rate and circulatory system involvement rate in the low load group were higher than those in the negative group(P<0.05). PLT counts in the high load group were significantly lower than those in the negative group, and the levels of GGT, TBIL, CK-MB, LDH, TG, SF, and organ involvement were significantly higher than those in the negative group. The levels of CK, LDH, SF and the number of organ involvement in the high load group were significantly higher than those in the low load group. The levels of GGT and TBIL in low load group were significantly higher than those in negative group. In terms of treatment, the proportion of blood purification therapy in the high and low load group was significantly higher than that in the negative group(P<0.01). ROC curve analysis showed that the best cut-off values of PLT, LDH, TG and SF were 49.5, 1139, 3.12 and 1812, respectively. The appellate laboratory indicators were dichotomized according to the cut-off value, and the differential clinical symptoms were included in the Cox regression model. Univariate analysis showed that LDH>1139 U/L, SF>1812 μg/L, dysfunction of central nervous system, number of organ damage, DIC and no blood purification therapy were the risk factors affecting the prognosis of children (P<0.05); Multivariate analysis shows that PLT≤49.5×109/L and dysfunction of central nervous system were risk factors affecting the prognosis of children (P<0.05). Survival analysis showed that there was no significant difference in the survival rate among the three groups.@*CONCLUSION@#The incidence of adverse prognostic factors in children with HLH in the EBV-DNA high load group is higher, and there is no significant difference in the survival rate of the three groups after blood purification therapy. Therefore, early identification and application of blood purification therapy is of great significance for children with HLH in the high load group.

The Prognostic Role of Circulating Epstein-Barr Virus DNA Copy Number in Angioimmunoblastic T-Cell Lymphoma Treated with Dose-Adjusted EPOCH / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Determine the frequency and prognostic value of circulating Epstein-Barr virus (EBV) DNA copy number in angioimmunoblastic T-cell lymphoma (AITL) patients who were treated with dose-adjusted etoposide, prednisone, vincristine, cyclophosphamide and doxorubicin (DA-EPOCH) regimens. MATERIALS AND METHODS: Sixty newly-diagnosed AITL patients were retrospectively enrolled in the present study. All patients were treated with DA-EPOCH regimen. RESULTS: Twenty-two subjects (36.7%) had a EBV DNA-positive test at diagnosis. EBV DNA‒positive patients were associated with lower lymphocyte-monocyte ratio (p=0.024). Median follow-up was 40 months (range, 14 to 100 months). The overall response rate for all the 60 AITL patents were 71.7% (95% confidence interval [CI], 58.6 to 82.5) with 3-year progressive-free survival (PFS) rate of 30.9%±6.1% and overall survival (OS) rate of 60.1%±6.6%. Not only did PFS estimation differ between the EBV DNA‒positive and EBV DNA‒negative group (hazard ratio [HR], 2.24; 95% CI, 1.15 to 4.35; p=0.006), but also worse OS was observed in the pretreatment EBV DNA‒positive group than in the EBV DNA‒negative group (HR, 2.74; 95% CI, 1.22 to 6.19; p=0.006). EBV DNA test positivity was independent prognostic marker for both PFS (HR, 2.17; 95% CI, 1.17 to 4.00; p=0.014) and OS (HR, 3.24; 95% CI, 1.48 to 7.11; p=0.004) after adjusting International Prognostic Index and prognostic index for AITL score. Reduction in EBV copies was significantly associated with therapy-response. CONCLUSION: Circulating EBV DNA level was an important prognostic and monitoring marker for AITL patients who treated with DA-EPOCH regimens which cannot improve outcomes for AITL patients.

Performance of Hepatitis B Core-Related Antigen Versus Hepatitis B Surface Antigen and Hepatitis B Virus DNA in Predicting HBeAg-positive and HBeAg-negative Chronic Hepatitis

BACKGROUND: We examined changes in hepatitis B core-related antigen (HBcrAg) during the four sequential phases of chronic hepatitis B virus (HBV) infection: hepatitis B e antigen (HBeAg)-positive chronic infection (EPCI) and hepatitis (EPCH), followed by HBeAg-negative chronic infection (ENCI) and hepatitis (ENCH). We compared the performance of serum HBcrAg, hepatitis B surface antigen (HBsAg), and HBV DNA in predicting EPCH and ENCH. METHODS: We enrolled 492 consecutive patients: 49 with EPCI, 243 with EPCH, 101 with ENCI, and 99 with ENCH. HBcrAg was detected by chemiluminescent enzyme immunoassays. HBsAg and HBeAg were detected by chemiluminescent microparticle immunoassays. HBV DNA was detected by real-time PCR. Predictive performance of HBcrAg, HBsAg, and HBV DNA was evaluated using ROC curves. RESULTS: Areas under ROC curves (AUCs) of HBcrAg, HBsAg, and HBV DNA for predicting EPCH were 0.738, 0.812, and 0.717, respectively; optimal cutoffs were ≤1.43×105 kU/mL, ≤1.89×104 IU/mL, and ≤3.97×107 IU/mL, with sensitivities and specificities of 66.3% and 77.6%, 65.0% and 93.9%, and 60.5% and 79.6%, respectively. AUCs of HBcrAg, HBsAg, and HBV DNA for predicting ENCH were 0.887, 0.581, and 0.978, respectively; optimal cutoffs were >26.8 kU/mL, >2.29×102 IU/mL, and >8.75×103 IU/mL, with sensitivities and specificities of 72.7% and 95.1%, 86.9% and 39.6%, and 89.9% and 92.1%, respectively. CONCLUSIONS: HBsAg and HBV DNA were the best predictors of EPCH and ENCH, respectively. HBcrAg is an important surrogate marker for predicting EPCH and ENCH.

Evaluation of the usefulness of a device for Human Papilloma Virus DNA collection and reservation in self-collected cervicovaginal samples stored dry in women with cervical dysplasia, Bogotá, Colombia / Evaluación de la utilidad de un dispositivo para toma y preservación del ADN del virus del papiloma humano de muestras cérvico-vaginales autorrecolectadas y almacenadas en seco en mujeres con displasia cervical, Bogotá, Colombia

ABSTRACT Objective: To evaluate the usefulness of a device for collecting and preserving human papilloma virus (HPV) DNA in self-collected vaginal samples stored dry during 14 days. Materials and methods: Diagnostic concordance pilot study that included non-pregnant women over 25 years of age with a biopsy-confirmed result of cervical intraepithelial neoplasia (CIN) grade 1 or more, coming to two referral centres in Bogotá, Colombia. Women with a history of total hysterectomy were excluded. Convenience sampling was used. The device uses real-time PCR (polymerase chain reaction) for DNA detection. Sociodemographic and clinical variables were measured, as well as the results of the test when the sample was collected by the patient and when it was collected by the physician, and the amount of DNA in the samples taken and processed on day 1, and in those processed on day 14, using Ct thresholds. Descriptive statistics were applied. Overall concordance was estimated by means of the kappa coefficient and mean differences in DNA amount. Materials and methods: Diagnostic concordance pilot study that included non-pregnant women over 25 years of age with a biopsy-confirmed result of cervical intraepithelial neoplasia (CIN) grade 1 or more, coming to two referral centres in Bogotá, Colombia. Women with a history of total hysterectomy were excluded. Convenience sampling was used. The device uses real-time PCR (polymerase chain reaction) for DNA detection. Sociodemographic and clinical variables were measured, as well as the results of the test when the sample was collected by the patient and when it was collected by the physician, and the amount of DNA in the samples taken and processed on day 1, and in those processed on day 14, using Ct thresholds. Descriptive statistics were applied. Overall concordance was estimated by means of the kappa coefficient and mean differences in DNA amount. Results: A kappa coefficient of 0.84 (95% CI: 0.71-0.96) was found for concordance in high-risk HPV detection between the self-collected cervicovaginal sample and the sample taken by the clinician. There were no differences in terms of the amount of viral DNA between day 1 and day 14 (DM -0.34 cycles; 95% CI: - 2.29 to 1.61). Conclusion: Self-collected vaginal samples using the storage device are reliable for high-risk HPV detection in patients with cervical dysplasia, and preserve viral DNA for 14 days if stored dry at room temperature. Confirmation studies in the general population are required.

RESUMEN Objetivo: Evaluar la utilidad de un dispositivo para toma y preservación del DNA del virus del papiloma humano (VPH) de muestras vaginales recolectadas por autotoma y almacenadas en seco durante 14 días. Materiales y métodos: Estudio piloto de concordancia diagnóstica. Se incluyeron mujeres mayores de 24 años no gestantes con un resultado de neoplasia intraepitelial cervical (NIC) grado 1 o más, confirmado por biopsia en dos instituciones de referencia en Bogotá, Colombia. Se excluyeron mujeres con antecedente de histerectomía total. Se realizó un muestreo por conveniencia. El dispositivo utiliza PCR (reacción en cadena de la polimerasa) en tiempo real para detección del ADN. Se midieron variables sociodemográficas y clínicas, así como el resultado de la prueba por autotoma y tomada por el médico, y la cantidad de ADN de las muestras tomadas el día 1 procesadas ese día, y el día 14, por medio del Ct umbral. Se realizó estadística descriptiva. Se calculó la concordancia global por medio del índice de kappa ponderado y la diferencia de medias de la cantidad de ADN. Resultados: La concordancia en la detección de VPH de alto riesgo mostró un kappa = 0,84 (IC 95 %: 0,71-0,96) entre la muestra cervicovaginal recolectada por autotoma frente a la muestra cervical recolectada por el médico. No hubo diferencias en la cantidad de ADN viral entre el día 1 y el 14 (DM -0,34 ciclos; IC 95 %: -2,29 a 1,61). Conclusión: Las muestras vaginales recolectadas por autotoma usando el dispositivo de almacenamiento son confiables para la detección de VPH de alto riesgo en pacientes con displasia cervical, y preservan el ADN viral por 14 días si se almacenan en seco a temperatura ambiente. Se requieren estudios en población general para poder confirmar.

Diagnostic value of lymph node EBV-DNA detection in cervical lymph node metastasis of nasopharyngeal carcinoma / 国际肿瘤学杂志

Objective To evaluate the diagnostic value of lymph node fine-needle aspiration (FNA)Epstein-Barr virus (EBV)-DNA concentration detection in nasopharyngeal carcinoma (NPC) cervical lymph node metastasis.Methods From August to December 2016,36 cases of NPC and 9 cases of other tumors (not correlated with EBV infection) were enrolled in this study at the Sun Yat-sen University Cancer Center.All patients received magnetic resonance images (MRI),plasma and cervical lymph node FNA EBV-DNA detection.Results The median concentration of EBV-DNA in FNA fluid (1.39 × 105 copies/ml) in cervical lymph node metastasis was significantly higher than that in plasma (2.00 × 103 copies/ml),with a significant difference (x2 =16.723,P =0.004).The diagnosis sensitivity,specificity,accuracy of the lymph node FNA fluid of EBV-DNA were 86.2％ (25/29),71.4％ (10/14) and 81.4％ (35/43) respectively,which were better than those of MRI [72.4％ (21/29),50.0％ (7/14) and 65.1％ (28/43) respectively] and plasma EBV-DNA [55.2％ (16/29),71.4％ (10/14) and 60.5％ (26/43) respectively].The area under the curve (AUC) of level Ⅰ b cervical lymph node metastasis was calculated,and FNA fluid EBV-DNA (AUC =0.688)was better than MRI (AUC =0.583),with a significant difference (Z =2.476,P =0.008).The EBV-DNA concentration in FNA fluid in cervical lymph node metastasis of patients with other tumors (no correlated with EBV infection) was 0 copy/ml.Conclusion FNA fluid EBV-DNA may improve the diagnostic sensitivity of cervical lymph node metastasis in nasopharyngeal carcinoma,and help to explore the clinical target volume neck nodes at level Ⅰ b cervical lymph node in radiotherapy.

The infection process of BK virus in transplant recipients after renal transplantation / 中华泌尿外科杂志

Objective To investigate the characteristics and manifestations of the different stages of BK virus infection in the recipients after renal transplantation.Methods A retrospective survey from January 2015 to December 2016 was done in our hospital.A total 135 recipients were included and accepted BK virus detection in 1,3,6,9,12,15 months respectively after renal transplantation.The prevalence of decoy cell,BK virus DNA load in urine and BK virus DNA load in blood was 56 cases (41.5％),9 cases (43.7％) and 30 cases (22.2％),5 cases of BK vims nephropathy confirmed by pathological biopsy (3.7％).At the same time,51 cases (37.8％) were combined with decoy cells and virus DNA load in urine.Positive decoy cells and negative BK virus DNA load in urine was 5 cases,and Positive BK virus DNA load in urine and negative decoy cells was 8 cases.The recipients were divided into positive group of urine decoy cell,positive group of urinary BK virus DNA load,and positive group of blood BK virus DNA load.Statistical correlation analysis was conducted on the laboratory test results of the 3 groups.Results The positive group of blood BK virus DNA load were detected the high level urine decoy cell count [median of 23/10HPF(2-48/10HPF)] and high level of urinary BK virus DNA load [4.52 × 106 copies/ml (6.51 × 103-7.89 × 109 copies/ml)],significantly higher than the positive group of decoy cells [8/10HPF(2-40/10HPF)] and the positive group of urine BK virus DNA load [4.56 × 105 copies/ml(5.62 × 103-7.89 ×109 copies/ml)] (P ＜ 0.05).The decoy cell count and urine DNA load has a significant linear correlation in viruria recipients,and the urinary BK DNA load and blood BK virus DNA load has the same significant 0.939 and 0.702 in 3 months,0.969 and 0.910 in 6 months,0.782 and 0.766 in 9 months,0.898 and 0.615 in 12 months after renal transplantation.Conclusions There is a linear correlation between decoy cell in urine,viruria and viremia,suggesting that the infection of BK virus in kidney transplant recipients is a continuous process.linear correlation in viremia recipients(P ＜ 0.05).The correlation coefficients at different time points were

Relationship between the HBsAg-positive infection status of mothers and the non/low-response to hepatitis B vaccine of their infants / 中华流行病学杂志

Objective To explore the relationship between the status of HBsAg-positive infection of mothers and the non/low-response to hepatitis B vaccine of their infants.Methods A total of 225 pairs of mothers and their infants were recruited in our cohort from June 2011 to July 2013.Infants were given three doses of hepatitis B vaccine at hour 24,first month and month 6th respectively and were followed up for one year after birth.HBV serological markers and HBV DNA in the peripheral blood of both mothers and infants were detected by Electro-chemiluminescence immunoassay and fluorescence quantitative Polymerase Chain Reaction.Results Six HBV infection models were detected in HBsAg-positive mothers,and "HBsAg (+),HBeAg (+),anti-HBc (+)" (model one) and "HBsAg (+),anti-HBe (+),anti-HBc (+)" (model two) accounted for 92.5％ (208/225) of all the models.Rate of non/low-response to hepatitis B vaccine in infants born to mothers in model one was lower than those in model two,the differences are statistically significant (x2=4.80,P=0.029).The rate of non/low-response to hepatitis B vaccine in infants showed a downward trend with the rising of HBeAg level in their mothers (x2=4.86,P=0.028).Results from the unconditional logistic regression analysis showed that the HBeAg of the HBsAg-positive mothers was significantly correlated with the low risk of non/low-response to hepatitis B vaccine in infants (OR=0.598,95％CI:0.378-0.947).The positive rate of serum HBV DNA in HBsAg-positive mothers was 54.2％,while the rate of non/low-response to hepatitis B vaccine in infants born to HBV DNA positive mothers was similar to those infants born to HBV DNA negative mothers (X2=0.22,P=0.640).Conclusions "HBsAg (+),HBeAg (+),anti-HBc (+)" and "HBsAg (+),anti-HBe(+),anti-HBc (+)" were the common models seen in HBsAg-positive mothers,and the rate of non/low-response to hepatitis B vaccine was different between the two models.HBeAg of HBsAg-positive mothers might have positive effects on the immune response to hepatitis B vaccine in infants but the mechanisms remained not clear.HBV DNA of the HBsAg-positive mothers did not seem to be correlated with the immune response to hepatitis B vaccine in infants.

Relationship between the HBsAg-positive infection status of mothers and the non/low-response to hepatitis B vaccine of their infants / 中华流行病学杂志

Objective To explore the relationship between the status of HBsAg-positive infection of mothers and the non/low-response to hepatitis B vaccine of their infants.Methods A total of 225 pairs of mothers and their infants were recruited in our cohort from June 2011 to July 2013.Infants were given three doses of hepatitis B vaccine at hour 24,first month and month 6th respectively and were followed up for one year after birth.HBV serological markers and HBV DNA in the peripheral blood of both mothers and infants were detected by Electro-chemiluminescence immunoassay and fluorescence quantitative Polymerase Chain Reaction.Results Six HBV infection models were detected in HBsAg-positive mothers,and "HBsAg (+),HBeAg (+),anti-HBc (+)" (model one) and "HBsAg (+),anti-HBe (+),anti-HBc (+)" (model two) accounted for 92.5％ (208/225) of all the models.Rate of non/low-response to hepatitis B vaccine in infants born to mothers in model one was lower than those in model two,the differences are statistically significant (x2=4.80,P=0.029).The rate of non/low-response to hepatitis B vaccine in infants showed a downward trend with the rising of HBeAg level in their mothers (x2=4.86,P=0.028).Results from the unconditional logistic regression analysis showed that the HBeAg of the HBsAg-positive mothers was significantly correlated with the low risk of non/low-response to hepatitis B vaccine in infants (OR=0.598,95％CI:0.378-0.947).The positive rate of serum HBV DNA in HBsAg-positive mothers was 54.2％,while the rate of non/low-response to hepatitis B vaccine in infants born to HBV DNA positive mothers was similar to those infants born to HBV DNA negative mothers (X2=0.22,P=0.640).Conclusions "HBsAg (+),HBeAg (+),anti-HBc (+)" and "HBsAg (+),anti-HBe(+),anti-HBc (+)" were the common models seen in HBsAg-positive mothers,and the rate of non/low-response to hepatitis B vaccine was different between the two models.HBeAg of HBsAg-positive mothers might have positive effects on the immune response to hepatitis B vaccine in infants but the mechanisms remained not clear.HBV DNA of the HBsAg-positive mothers did not seem to be correlated with the immune response to hepatitis B vaccine in infants.

Expression of Epstein-Barr Virus DNA in Peripheral Blood Lymphocytes and Plasma in Patients with EBV-Associated Diseases / 现代检验医学杂志

Objective To explore the expression of Epstein-Barr virus DNA(EBV DNA)in the peripheral blood lymphocytes and plasma of patients with EBV-associated diseases.Methods The whole blood samples were collected from 112 patients with suspected EB virus infection diseases,including 14 cases of nasopharyngeal carcinoma(NPC),16 cases of infectious mononucleosis(IM),22 cases of lymphoma,23 cases of autoimmune disease,26 cases of upper respiratory tract infection, and 11 cases of abnormal liver function.The levels of EBV DNA in lymphocytes and plasma of the same sample were detec-ted by fluorescence quantitative PCR(FQ-PCR).Results The EBV DNA positive rates in lymphocytes and plasma of all 112 patients were 83.0%(93/112)and 27.7%(31/112)respectively,with statistically significant difference(χ2=60.02,P<0.01).The positive rate and the load of EB virus DNA in lymphocytes and plasma of 14 patients with nasopharyngeal car-cinoma(NPC)had no statistical difference(χ2=2.25,t=-1.04,all P>0.05).However,patients with lymphoma,infec-tious mononucleosis,upper respiratory tract infection,autoimmune disease or abnormal liver function,the positive rates and the concentration of EBV DNA in the plasma were dramatically lower than those in the peripheral blood lymphocytes,and the difference was statistically significant(χ2=4.17~15.06,all P<0.05;t=3.94~10.45,all P<0.01).Conclusion The detection of EB DNA in peripheral blood lymphocytes of non NPC patients by FQ-PCR might be better than that in plasma. There was no statistical difference between the detection of EBV DNA in lymphocytes and plasma of patients with nasopha-ryngeal carcinoma.Appropriate specimen type could be selected according to clinical consideration.

Establishment and Application of SYBR Green I Real-Time PCR Assay for Rapid Detection of Hepatitis B Virus DNA / 现代检验医学杂志

Objective To develop SYBR Green I real-time PCR assay for detection and identification of Hepatitis B virus. Methods Based on the sequences of Hepatitis B virus gp1 gene,primers were designed.The reaction assay and thermal cyc-ling profile were optimized.The positive standard was from recombinant clone.Both the developed assay and Zhejiang kuake biotechnology company’s assay were applied in 100 patients serum.Results The detection limit was between 5×102 copies/ml to 5×108 copies/ml with a good liner correlation and no cross reaction.The whole process just needed 2.5 h.Comparing with the company products,the sensitivity and specificity of the developed assay were 100% and 92.5% respectively.Con-clusion The established assay is rapid,simple,high sensitivity and specificity.It is not only valuable for the identification of Hepatitis B virus patients,but also provide accurate quantitative analysis for HBV patients.

Relationship between MDSC level and immune function and HBV-DNA replication level in patients with hepatitis B / 中国基层医药

Objective To investigate the relationship between myeloid-derived suppressor cell ( MDSC) levels and immune function and hepatitis B virus DNA ( HBV-DNA) replication level in patients with hepatitis B. Methods 96 patients with hepatitis B were included in the study.40 normal persons who received physical examina-tion in the hospital during the same period were included as the control group.The content of MDSC was detected by flow cytometry and HBV-DNA level was detected by real-time fluorescence quantitative PCR technique.Statistical analysis was performed.Results The content of MDCS in hepatitis B group (8.77 ±3.69)%was significantly high-er than (1.64 ±1.26)%in the control group (t=16.80,P107 IU/mL ( 90.91%) was significantly higher than that in HBsAg negative group (2.70%)(P<0.05).The content of MDSC in HBV DNA positive group[(10.82 ±3.26)%] was significantly higher than that in HBV DNA negative group[(5.84 ±2.16)%](t=8.42,P<0.05).Spearman's correlation analy-sis showed that the level of MDCS expression was positively correlated with the level of DNA replication (r=0.769, P<0.05).Conclusion The increased level of MDSC content can cause the immune suppression and is positively correlated with the degree of HBV DNA replication.

Correlation between two chemiluminescence based assays for quantification of hepatitis B surface antigen in patients with chronic hepatitis B infection.

Purpose: Hepatitis B surface Antigen (HBsAg) is the hallmark in diagnosing hepatitis B virus (HBV) infection. In India many commercial assays are available for detection of HBsAg but very few can measure it quantitatively. The present study presents the comparative evaluation of two methods and their correlation with serum HBsAg in chronic hepatitis B (CHB) patients. Materials and Methods: Consecutive patients of CHB were included and there HBsAg levels were measured by two methods: (i) Elecsys, Roche Diagnostics, a qualitative assay and (ii) Architect, Abbott Diagnostics, a quantitative assay. The HBV DNA was measured by real-time polymerase chain reaction (qPCR). Results: Total of 136 patients were included in the study and there was a signifi cant overall correlation between both the assays (correlation coeffi cient [r] = 0.83; P < 0.001). Assays correlated well with each other across all subgroups of CHB: treatment naïve (r = 0.73; P < 0.001, n = 32), on treatment (r = 0.56; P < 0.05, n = 104), hepatitis Be (HBe) antigen positive (r = 0.67; P < 0.001, n = 62) and anti-HBe positive (r = 0.61; P < 0.05, n = 74) group. On correlation with serum HBV DNA, Architect assay demonstrated good correlation (r = 0.73; P < 0.001, n = 136) as compared to the Elecsys assay (r = 0.27; P = 0.068, n = 136). Architect HBsAg QT assay (A1) also correlated well with HBV DNA in the treatment naïve group (r = 0.69; P < 0.001, n = 32). Conclusions: Our study hence proved that both the assays are comparable and a simple qualitative assay with in-house modifi cation can be used easily for quatitation of HBsAg in clinical samples.

Effect of preoperative antiviral therapy on HBV reactivation and postoperative liver function in perioperative patients with HBV-DNA-negative hepatocellular carcinoma / 中国生化药物杂志

Objective To explore the effect of preoperative antiviral therapy on hepatitis B virus ( HBV ) reactivation and postoperative liver function in perioperative patients with HBV-DNA-negative hepatocellular carcinoma(HCC).Methods 74 patients with preoperative HBV-DNA-negative scheduled which were analyzed.Patients were divided into two groups according to antiviral therapy or not:20 cases in antiviral treatment group received antiviral therapy for three days, 54 cases in non-antiviral teatment group did not receive antiviral therapy, and both groups received antiviral therapy after post-operative resuming to diets.The indicators of liver function and HBV-DNA levels were detected on pre-operative, post-operative 3rd and 7th day in two groups, and HBV-DNA-positive ( HBV-DNA>500 IU/mL) was defined as reactivation, conversely as inactivation.The indicators of liver function on pre-operative, post-operative 3rd and 7th day were compared between reactivation group and inactivation group.Results The reactivative rate was 21.6%(16/74) in all patients;27.7%(15/54) in pre-operative non-antiviral teatment group, 5.0%(1/20) in antiviral teatment group, and there was significant differences in reactivative rate between two groups ( P=0.035 ).The results of Logistic regression showed that pre-operative nonantiviral therapy was an independent risk factor of post-operative HBV reactivation (OR=13.952,95% confidence interval[CI]:1.358-143.379,P=0.027).The recovery of albumin (ALB) on post-operative 3rd, 7th days in antiviral treatment group was faster than those in nonantiviral treatment group, respectively (P=0.035,0.043).The recovery of ALB and alanine aminotransferase (ALT) on post-operative 7th day in reactivation group were slower than those in inactivation group, respectively (P=0.016, 0.048).Conclusion The pre-operative nonantiviral therapy is an independent risk factor of post-operative HBV reactivation in patients with HBV-DNA-negative HCC.The pre-operative antiviral therapy could inhibit post-operative HBV reactivation effectively and accelerate the post-operative recovery of liver function.

Detection of Human Hepatocarcinoma Cell Line PLC/PRF/5 Genome Hepatitis b Virus DNA Integration with Alu-PCR / 医学研究杂志

Objective In this research with the method of Alu -PCR we investigate the integration of hepatitis B virus ( HBV) DNA in human hepatocarcinoma cell line (PLC/PRF/5) genome DNA.Methods We at first extracted the genome DNA from PLC/PRF/5 cells, and then the potential integration fragments of HBV DNA and human genome DNA were amplified with according to the Alu -PCR after three rounds PCR .The Alu-PCR amplification products were observated with agarose gel electrophoresis , then integration positive electrophoresis bandings were chipped and purified for nucleic acid sequencing .At last he bioinformatics information was acquired by blast online.Results Through agarose gel electrophoresis after Alu -PCR amplification, we got four potential integration bindings , among which we got three integration sequences of HBV DNA in human genome DNA .These integration sequences could be individually located in the human chromosome of 03p21.31, 05p15.33, 12q13 and 12-q14.1.Conclusion With Alu-PCR we can accurately measure the integration of HBV DNA in human genome DNA , and Alu-PCR can be a a convenience and economic method in the study of HBV DNA ′s integration in human genome DNA .

Correlation Analysis of Maternal and Neonatal HBV Test Results / 现代检验医学杂志

Objective To investigate the risk factors of intrauterine hepatitis B virus (HBV)infection and its preventive measures.Methods 254 cases of pregnant women with positive HBsAg and their neonatuses from Shaanxi People’s Hospi-tal were selected as the research object.The serum of pregnant women were tested HBV markers and HBV-DNA,their neo-natal umbilical cord blood were only detected HBV markers.Results In 254 samples,the positive rates of HBsAg and HBeAg within their neonatal umbilical cord blood were 5.12% and 13.78% respectively;the positive rate of HBV infection in neonatal umbilical cord blood among maternal HBeAg-positive group was 53.33%,far higher than 3.61% in HBeAg-neg-ative group (χ2 =99.44,P <0.001);the positive rates of HBsAg and HBeAg in neonatal umbilical cord blood were elevated along with the increase of maternal HBV-DNA copy (Hc= 13.50,66.70;P < 0.001).The positive rate of HBV-DNA in pregnant women with HBeAg-positive group was 100%,far more than 19.59% in HBeAg-negative group (P <0.001).Con-clusion Both maternal positive HBeAg and HBV-DNA are risk factors for neonatal intrauterine HBV infection.Prenatal HBeAg screening is predominant,especially in some hospitals without real-time quantitative assay.An effective measure to reduce intrauterine HBV infection is to be pregnant when HBV DNA and HBeAg are turned into negative after HBV inter-vention and treatment for women in reproductive age.

Reacción en cadena de la polimerasa en tiempo real para la cuantificación del ADN del virus de la hepatitis B / Real-time polymerase chain reaction assay for Hepatitis B virus DNA quantification

Introducción: los niveles de ADN viral en muestras de suero son un marcador útil para monitorear la progresión de la enfermedad y la respuesta al tratamiento en pacientes con hepatitis B crónica; de ahí que se comercialicen estuches diagnósticos para esta función, con la desventaja de ser costosos. Objetivos: desarrollar y evaluar el desempeño analítico de un sistema de reacción en cadena de la polimerasa en tiempo real para la cuantificación del ADN del virus de la hepatitis B. Métodos: se utilizaron cebadores que amplifican un fragmento del gen C y sonda de hidrólisis en el equipo LightCycler 1.5. Se construyó una curva estándar y se evaluó su eficiencia. Se utilizaron 272 muestras de suero para ensayos de especificidad analítica y clínica, especificidad y exactitud genotípica, coeficientes de variación intraensayo e interensayo, comparación con un estuche comercial y con la reacción en cadena de la polimerasa cualitativa para el virus de la hepatitis B. Resultados: la curva estándar mostró excelente correlación lineal (r= -1) y valores muy bajos de error a lo largo de varias magnitudes de concentración de ADN diana. La especificidad analítica y clínica fue de 100 %, en tanto que al evaluar la especificidad y exactitud genotípica, se obtuvo que las diferencias entre los Log10 del valor obtenido y el de referencia eran inferiores a 0,5 Log10. El límite de detección por análisis de Probit se estimó en 16,41 UI/µL con un rango dinámico de cuantificación de hasta 10(8) UI/mL. El sistema mostró bajos coeficientes de variación intraensayo (0,16 a 1,45 %) e interensayo (0,9 a 2,62 %). La comparación con el estuche comercial artus HBV LC PCR kit mostró una correlación de r= 0,964 y r²= 0,929; con la reacción en cadena de la polimerasa cualitativa se confirmó la mayor sensibilidad y ventajas de la reacción en cadena de la polimerasa en tiempo real. Conclusiones: el ensayo cumple con los requisitos para la cuantificación del ADN del virus de la hepatitis B, que demuestra ser específico, sensible y reproducible. Su aplicación permitirá un mejor diagnóstico y seguimiento de los pacientes con hepatitis B crónica en Cuba.

Introduction: viral DNA levels in serum samples are a useful marker to monitor the disease progression and the treatment response in patients with chronic hepatitis B. Commercial kits for this purpose are available, but they are considerably expensive. Objectives: to evaluate the analytical performance of a real-time polymerase chain reaction (RT-PCR) assay for Hepatitis B virus DNA quantification. Methods: specific primers to the gene C and TaqMan chemistry in a LightCycler 1.5 equipment was used. A standard curve was made and evaluated. Two hundred and seventy-two serum samples were used to assess the clinical and analytical specificity, the genotypic accuracy and specificity, the intra-assay and interassay coefficients of variation and the comparison with a commercial assay and with the qualitative PCR. Results: the standard curve showed a strong linear correlation (r= -1) and low error values in the tested target DNA concentration. Analytical and clinical specificities were 100 %. Genotype accuracy and specificity showed that the differences between the results obtained by RT-PCR assay and those of the reference assay were less than 0.5 Log10. The 95% HBV DNA detection end-point assessed by Probit analysis was 16.41 IU/µL with a dynamic range of quantification of 10(8) IU/mL. Intra-assay and interassay coefficients of variation ranged from 0.16 to 1.45 % and 0.9 to 2.62 % respectively. The RT-PCR assay correlated well with those from a commercial assay (r= 0.964 and r²= 0.929) and with the HBV qualitative PCR, thus confirming its better sensitivity and advantages. Conclusions: the RT-PCR assay is well suited to monitoring HBV DNA levels showing to be sensitive, specific and reproducible. Its application in the clinical practice ensures a better diagnosis and management of patients with chronic hepatitis B in Cuba.

High prevalence of the simultaneous excretion of polyomaviruses JC and BK in the urine of HIV-infected patients without neurological symptoms in São Paulo, Brazil / Alta prevalência de excreção simultânea de poliomavírus JC e BK na urina de pacientes HIV+ sem sintomas neurológicos em São Paulo, Brasil

OBJECTIVE: To evaluate the prevalence of the urinary excretion of BKV and JCV in HIV-infected patients without neurological symptoms. METHODS: Urine samples from HIV-infected patients without neurological symptoms were tested for JC virus and BK virus by PCR. Samples were screened for the presence of polyomavirus with sets of primers complementary to the early region of JCV and BKV genome (AgT). The presence of JC virus or BK virus were confirmed by two other PCR assays using sets of primers complementary to the VP1 gene of each virus. Analysis of the data was performed by the Kruskal-Wallis test for numerical data and Pearson or Yates for categorical variables. RESULTS: A total of 75 patients were included in the study. The overall prevalence of polyomavirus DNA urinary shedding was 67/75 (89.3%). Only BKV DNA was detected in 14/75 (18.7%) urine samples, and only JCV DNA was detected in 11/75 (14.7%) samples. Both BKV and JCV DNA were present in 42/75 (56.0%) samples. CONCLUSION: In this study we found high rates of excretion of JCV, BKV, and simultaneous excretion in HIV+ patients. Also these results differ from the others available on the literature.

OBJETIVO: Avaliar a prevalência de excreção urinaria de vírus JC (VJC) e vírus BK (VBK) em pacientes HIV+ sem sintomas neurológicos. MÉTODOS: Amostras de urina de pacientes HIV+ sem sintomas neurológicos foram testados para a presença de VJC e VBK através da técnica de PCR. As amostras foram triadas para a presença de poliomavírus com par de primers complementares a região precoce do genoma do VBK e do VJC (AgT). A presença foi confirmada através de dois outros ensaios de PCR dirigidos a região do gene VP1 de ambos os vírus. A análise estatística foi realizada com auxílio do teste de Kruskal-Wallis para dados numéricos e Pearson ou Yater para variáveis categóricas. RESULTADOS: Ao todo foram inclusos no estudo 75 pacientes. A prevalência geral de excreção de poliomavírus na urina foi de 67/75 (89,3%). O DNA do vírus VBK foi detectado em 14/75 (18,7%) das amostras de urina, e o DNA do VJC foi detectado em 11/75 (14,7%) das amostras testadas. Ambos os vírus estavam presentes simultaneamente em 42/75 (56%) das amostras de urina. CONCLUSÃO: Encontramos, no presente estudo, uma alta taxa de excreção de VJC, VBK e excreção simultânea em pacientes HIV+. Ainda, esses resultados diferem de outros disponíveis na literatura.

Serum hepatitis B surface antigen levels correlate with high serum HBV DNA levels in patients with chronic hepatitis B: A cross-sectional study.

Purpose : The hallmark of chronic hepatitis B (CHB) infection is the presence of hepatitis B surface antigen (HBsAg) positivity for at least 6 months. Recently, serum levels of HBsAg have been compared with serum HBV DNA as a surrogate marker to monitor CHB patients. However, data correlating these two markers are scarce. Hence, the present study was done to correlate HBV DNA with HBsAg in CHB patients. Materials and Methods: Consecutive patients of CHB were included. HBV DNA was measured by real-time polymerase chain reaction (PCR). Serum HBsAg was measured by Architect HBsAg. Results: Of the 198 patients enrolled, 166 fulfilled the inclusion criteria (mean age 43 ± 14 years, 87% males) and the median HBV DNA was 1.7 × 10 3 (range 6.0-1.1 × 10 8 ) IU/ml. Median HBsAg was 8.7 × 10 3 (range 5.0-3.2 × 10 5) IU/ml. Overall correlation between HBV DNA and HBsAg was weak but significant (Spearman ρ = 0.443, P < 0.01). Correlation in HBe antigen-positive group was better (ρ = 0.402, P < 0.01) in comparison to HBe antigen-negative group (ρ = 0.193 P = 0.05). Good correlation existed in treatment-naïve group (ρ = 0.538, P < 0.01) .Correlation was regardless of normal or raised alanine transaminase (ALT). Eighty (48%) patients had high HBV DNA (≥2000 IU/ml). Correlation in high DNA group was significant (P < 0.01). The best cut-off of HBsAg for diagnosing high DNA is 3.36 ×10 3 IU/ml. Conclusions: Serum HBsAg correlates with HBV DNA in CHB patients, especially in high serum HBV DNA, HBe antigen-positive and treatment-naïve group. HBsAg levels can be used for predicting high serum HBV DNA levels.

Clinical Characteristics of Occult HBV Infection and Impact on Treatment Response in Patients with Chronic Hepatitis C / 대한내과학회지

BACKGROUND/AIMS: The prevalence of occult HBV infection (OBI) in patients with chronic hepatitis C (CHC) in Korea has not been reported. Additionally, it is unclear whether OBI influences treatment outcome in CHC patients. We investigated the prevalence of OBI and its impact on treatment outcome in patients with CHC. METHODS: Seventy-six patients with CHC were enrolled and treated with pegylated or conventional interferon and ribavirin. Hepatitis B virus (HBV) DNA was detected by nested polymerase chain reaction. RESULTS: Among the 68 patients who completed treatment and follow-up, HBV DNA was detected in serum from nine (13.2%) patients, liver tissue from 10 (14.7%), and serum or liver tissue from 15 (22.1%). OBI was diagnosed in nine (12.7%) control subjects. No difference in the prevalence of OBI between patients with CHC and controls was observed (13.2 vs. 12.0%; p = 0.92). No significant differences in age, sex, genotype 1 frequency, amount of hepatitis C virus RNA, anti-hepatitis B surface antigen/anti-hepatitis B core-IgG seropositivity, staging, or histology grading were observed in patients with or without HBV DNA. Sustained virological response was achieved in 73.3% of patients with OBI and 83.0% without OBI (p = 0.46). CONCLUSIONS: These results demonstrate that a significant proportion of patients with CHC have occult HBV infection and that OBI does not affect treatment outcome in patients with CHC.

Infecção pelo papilomavírus humano: etiopatogenia, biologia molecular e manifestações clínicas / Human papillomavirus infection: etiopathogenesis, molecular biology and clinical manifestations

O papilomavírus humano (HPV) é um vírus DNA que apresenta tropismo por células epiteliais, causando infecções na pele e nas mucosas. A replicação do HPV ocorre no núcleo das células escamosas e o seu ciclo de vida é diretamente relacionado ao programa de diferenciação da célula hospedeira. Até o momento, foram completamente caracterizados cerca de 100 tipos diferentes de HPVs e há um grande número adicional de tipos ainda não sequenciados. Além de ser o responsável por lesões benignas de pele e mucosas, o HPV também está envolvido no desenvolvimento de diversos tumores cutaneomucosos: doença de Bowen, cânceres de pele não melanoma e carcinomas genitais. Esta revisão aborda as características do HPV, quadros cutâneos e mucosos benignos e malignos causados por ele e os principais métodos empregados em sua detecção e tipagem.

Human papillomavirus (HPV) is a DNA virus that presents tropism for epithelial cells, causing infections of the skin and mucous membranes. Replication of HPV occurs in the nuclei of squamous cells and its life cycle is directly related to the differentiation program of the host cell. To date, nearly 100 different types of HPV have been characterized and there is a large number of other types that have not been sequenced yet. Besides being responsible for benign lesions of the skin and mucous membranes, HPV is also involved in the development of various mucocutaneous tumors: Bowen's disease, non-melanoma skin cancers and genital carcinomas. This review discusses the characteristics of HPV, malignant and benign mucous and skin manifestations caused by HPV, besides the main methods of detection and typing of the virus.