Hepatitis B virus,hepatitis C virus

Medical workers should have anti-HBV antibody to protect HBV infection in the hospital. If they have not anti-HBV antibody, they should receive HBV vaccination. HBV vaccination program is as follows: 10 micrograms, s.c., 0, 1, 6 months. In case of HBV contamination, 1,000 IU hepatitis B immune globulin(HBIG) and/or 10 micrograms HB vaccine should be administered judging from HBV markers of contaminated subjects and HBV load of patients. The HCV vaccine is not available. In case of HCV contamination, it is unnecessary to treat just after accident. If acute hepatitis C is evolved in those subjects during follow-up, it is recommended to treat with interferon. Eradication of HCV by interferon among patients with acute hepatitis C will be almost 100%.     

Partitioning of hepatitis C virus during Cohn-Oncley fractionation of plasma

Because of concern about the safety of immune globulins with respect to transmission of hepatitis C, the partitioning of hepatitis C virus (HCV) during alcohol fractionation of a plasma pool prepared exclusively from anti-HCV-reactive donations was examined. Quantitation of HCV RNA was accomplished by nested polymerase chain reaction (PCR) at limiting dilutions. One PCR unit was arbitrarily defined as the minimum amount of HCV RNA from which an amplified product could be detected. The starting plasma pool contained 1.4 x 10(5) PCR units per mL. Most of the HCV RNA was found in cryoprecipitate and in Cohn fractions I and III, but it was also detected in fraction II, which is used for immunoglobulin G preparations. A 3.4-percent solution of IgG prepared from this fraction II contained 30 PCR units per mL. The fractionation process leading to immune globulin resulted in overall reduction in HCV RNA by a factor of 4.7 x 10(4). Although the presence of HCV RNA in the final product does not necessarily imply the presence of infectious virus, this work suggests that the safety of immune globulins with respect to HCV transmission is not due solely to the partitioning of HCV away from the immunoglobulin fraction.     

Inactivation of hepatitis A virus in plasma products by vapor heating

BACKGROUND: The transmission of hepatitis A virus (HAV) has been associated with the use of a number of solvent/detergent-treated factor VIII concentrates and possibly a factor IX concentrate. These reports have emphasized the necessity of using virus-inactivation methods for plasma products that are capable of inactivating nonenveloped viruses such as HAV. STUDY DESIGN AND METHODS: A simple, highly accurate titration procedure for HAV, which allows extensive kinetic investigations of virus-inactivation procedures, has been developed. This system has now been used to evaluate the efficacy of vapor heating in inactivating HAV after the addition of the virus to a range of human plasma products. RESULTS: It was demonstrated that HAV was significantly more thermostable than other picornaviruses, which reinforced the fact that such viruses cannot be used as model viruses for HAV-inactivation studies. A one-step vapor-heating procedure was demonstrated to inactivate between 5.9 and > 6.3 log10 of HAV in different products. A two-step vapor-heating procedure had the capacity to inactivate between > 8.7 and > 10.4 log10 of HAV. Both procedures were more effective in inactivating HAV than was the pasteurization procedure used for virus inactivation in human albumin solutions. CONCLUSION: These data demonstrate the efficacy of vapor heating in inactivating high-titer HAV after the spiking of plasma products with virus. This study confirms and explains the results of controlled clinical trials and long-term clinical usage with respect to the lack of HAV transmission by such vapor-heated products.     

Residual risk of transfusion-transmitted human immunodeficiency virus, hepatitis C virus, and hepatitis B virus infections in Italy

BACKGROUND: Estimating the risk of transfusion-transmitted infections (TTIs) is essential for monitoring blood safety. The residual risk of TTI was estimated for nearly 90 percent of the blood supply in Italy. STUDY DESIGN AND METHODS: Data were analyzed from 1,079,281 repeat donors, corresponding to 5,361,000 donations made in blood transfusion centers throughout Italy in the period 1999 through 2001. The residual risk of transfusion-transmitted human immunodeficiency virus (HIV), hepatitis C virus (HCV), and hepatitis B virus (HBV) infections was estimated with the incidence rate-window period model. The denominator for the incidence rate (i.e., the number of person-years at risk) was estimated on a sample of 5850 donors. RESULTS: The risk of an infectious donation entering the blood supply, per 1 million donations, was 1.91 (probable range, 0.52-3.32) for HIV, 16.74 (9.57-24.01) for HCV, and 69.16 (43.12-102.70) for total HBV (adjusted for vaccination and hepatitis B surface antigen transience). CONCLUSION: In Italy, the estimated residual risk of TTI is apparently low, particularly for HIV infection. Although the estimated risks are higher for HCV and HBV, the introduction of mandatory viral detection tests for HCV in 2002 should account for an 80 percent reduction in the HCV risk. Moreover, the ongoing HBV vaccination program will contribute to reducing the risk of transfusion-transmitted HBV.     

Neopterin levels during the early phase of human immunodeficiency virus, hepatitis C virus, or hepatitis B virus infection

BACKGROUND: A study was conducted to assess the diagnostic sensitivity of neopterin screening of blood donors with regard to the detection of window-phase specimens of human immunodeficiency virus (HIV), hepatitis C virus (HCV), and hepatitis B virus (HBV) infection. STUDY DESIGN AND METHODS: In total, 1002 diagnostic window-phase specimens from 98 seroconversion panels (29 HIV-1, 52 HCV, and 17 HBV) were analyzed with viral antigen detection, viral nucleic acid amplification testing (NAT), and neopterin quantitation assays. The study was completed by the analysis of 92 anti-hepatitis B core antigen (HBc)-reactive and 103 alanine aminotransferase (ALT)-elevated blood donor specimens. RESULTS: A significant association between elevated neopterin concentrations and the very early phase of HIV-1 infection was found. No significant correlation could be observed between neopterin levels and the early phase of HCV or HBV infection. Neopterin concentration was not increased in specimens from blood donors with anti-HBc reactivity or ALT elevation. CONCLUSIONS: Neopterin screening of blood donors may identify window-phase cases of HIV, but not of HCV or HBV infection. The diagnostic sensitivity of neopterin screening during the HIV window phase is similar to that of the p24 antigen test. With the introduction of viral NATs in blood screening, there is no additional benefit of neopterin screening with regard to the three blood-borne viruses HIV, HCV, and HBV. Acute phases of other infectious agents, however, have been reported to be detected by neopterin enzyme-linked immunosorbent assays.     

乙型肝炎病毒DNA在低温储存血浆中稳定性的研究

目的了解-20℃条件下储存30d后血浆中乙型肝炎病毒(HBV)脱氧核糖核酸(DNA)的稳定性。方法收集载量103~107 copy/mL的30份乙型肝炎患者血浆,并分为低载量组、中载量组、高载量组。标本采集后立即分离血浆并进行基线(0d)载量检测,其余血浆于-20℃存储,分别在第1、3、7、14、21和30天进行实时荧光定量测定。结果 (1)将30份样本进行不同时间点的配对t检验,载量差异均无统计学意义:第1天与第0天比较(t=-0.327,P=0.746);第3天与第0天比较(t=-0.718,P=0.479);第7天与第0天比较(t=-0.682,P=0.500);第14天与第0天比较(t=-1.072,P=0.292);第21天与第0天比较(t=-0.818,P=0.420);第30天与第0天比较(t=0.635,P=0.530)。(2)分组比较中,第30天与第0天比较,HBV DNA载量差异无统计学意义:低载量组P=0.984;中载量组P=0.708;高载量组P=0.120。结论 -20℃储存30d,血浆标本HBV DNA载量未出现连续下降或上升趋势,且不同时间点与第0天的基线载量比较差异无统计学意义。     

Risk factors and seroprevalence of hepatitis B virus, hepatitis C virus, and human immunodeficiency virus infection in uzbekistan.

OBJECTIVES: The aim of this study was to elucidate the seroprevalence of hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) infection in Uzbekistan and to explore whether there is a correlation between those blood-borne agents and socioeconomic risk factors. METHODS: One thousand nine hundred and eighteen subjects were studied. The subjects were divided into a low-risk group, a high-risk group and a patient group. Sera were tested for HBV surface antigen (HBsAg), anti-HCV, and anti-HIV. RESULTS: The seroprevalence of HBsAg, anti-HCV, and anti-HIV in the general population was 13.3, 13.1 and 0%, respectively. The anti-HCV infection rate was significantly higher in intravenous drug users (62.7%) than in prostitutes (9.2%), homosexuals (11.1%), and medical laboratory employees (12.5%) (p < 0.01). In the low-risk group, positivity for anti-HCV increased with age from 2.2% in the 15- to 20-year-olds up to the highest rate of 17.6% in the 31- to 40-year-olds; the positivity then decreased to 0% in the group over 60 years of age. In the high-risk group, the positivity for anti-HCV in the age groups under 40 years was approximately 30% and significantly higher than in the low-risk group (p < 0.01). Risk factors for transmission of HCV were medical treatment in the low-risk group, drug abuse in the high-risk group, and both in the patient group. CONCLUSIONS: This study demonstrates that the seroprevalence of HBV and HCV infection is high, whereas HIV infection is yet uncommon in Uzbekistan.     

四川地区供血浆人群隐匿性乙型肝炎病毒感染的研究

目的了解四川地区供血浆人群隐匿性乙型肝炎病毒感染(OBI)情况,分析现行标准下原料血浆的HBV残余风险。方法采用实时荧光PCR和ELISA法,对四川地区10个单采血浆站2007年7月～2009年7月筛查合格的56 620名次供浆者的135 542份原料血浆标本作HBV NAT和HBsAg同步筛查,对筛查阳性的标本以进口试剂复核、作中和试验及HBV DNA定量测定,并对阳性血浆的供血浆者追踪分析,确认OBI标本。结果共检出HBV DNA阳性12份(9人),四川地区原料血浆HBV DNA检出率为0.008 9%(12/135 542);HBV DNA载量均<1 000 IU/ml;采用国产ELISA试剂检测该12份标本HBsAg均为阴性,而用进口试剂检测并经中和试验确认了其中5份(3人)为HBsAg阳性,其余7份(6人)为OBI血浆,血清学模式均为HBsAg-/HBeAg-/HBcAb+;对供血浆者2～9个月的追踪分析显示ELISA和NAT检测结果无变化,与筛查结果一致。结论 HBsAg阴性供血浆人群中存在OBI感染者,四川供浆人群中OBI感染率为0.010 6%(6/56 620),低于现有报道的我国无偿献血者的OBI感染率;按照现行要求采用ELISA法检测原料血浆,灵敏度较低的国产试剂检测HBsAg的残余风险(0.0089%)高于进口试剂(0.005 2%)。增加HBV NAT能有效降低原料血浆OBI的漏检风险。     

Prevalence and trends of human immunodeficiency virus, hepatitis B virus, and hepatitis C virus among blood donors in Iran, 2004 through 2007

BACKGROUND: Evaluation and monitoring the prevalence of transfusion-transmissible viral infections in blood donors is a valuable index of donor selection and blood safety. This study analyzed the trends of blood-borne infections among Iranian blood donations during 4 years.
STUDY DESIGN AND METHODS: Viral screening results of 6,499,851 allogeneic donations from 2004 through 2007 were analyzed. All donations were screened for hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV), and syphilis. The prevalence of HBV, HCV, and HIV infections per 100,000 donations and 95% confidence interval was calculated. The p value was estimated by chi-square test.
RESULTS: The prevalences of HBV, HCV, and HIV decreased during the 4-year study from 2004 through 2007. The overall prevalence was 0.56% for HBV, 0.004% for HIV, and 0.13% for HCV. There was a significant and impressive decrease in hepatitis B surface antigen prevalence from 0.73% in 2004 to 0.41% in 2007. The prevalence of HIV appeared to have decreased from 0.005% in 2004 to 0.004% in 2007 although the decrease was not significant. HCV prevalence showed a slight decline in blood donations from 0.14% in 2005 to 0.12% in 2007.
CONCLUSION: The trends of transfusion-transmitted infection prevalence in Iranian blood donations suggest that most of the safety measures employed in recent years in Iran have been effective.     

血浆中丙型肝炎病毒核酸在不同保存条件下的稳定性

目的探讨不同保存条件和时间对丙型肝炎病毒(HCV)RNA稳定性的影响。方法将22份血浆样本置于-20℃保存4周,4℃保存12d,室温(20~25℃)保存7d,反复冻融5次,采用荧光定量聚合酶链反应测定HCV RNA水平,考察HCV RNA稳定性。结果样本在-20℃保存4周HCV RNA平均下降幅度小于或等于0.14lg,±95%CI≤0.20lg;4℃保存12dHCV RNA平均下降幅度小于或等于0.39lg,4℃保存12d时95%CI下限为-0.50lg,12d时±95%CI≤0.24lg;室温保存7dHCV RNA平均下降幅度小于或等于0.40lg,室温保存7d处理组95%CI下限为-0.53lg,7d时±95%CI≤0.29lg;反复冻融5次HCV RNA平均下降幅度小于或等于0.10lg,±95%CI≤0.15lg。结论血浆样本在-20℃保存4周、4℃保存9d、室温保存5d和反复冻融5次时,对HCV RNA水平影响不明显,但4℃保存12d或室温保存7d后,RNA水平显著降低,所以应避免在这种条件和时间下保存和运输血浆。     

Prevalence and risk factors of hepatitis C virus, hepatitis B virus, and human immunodeficiency virus in multiply transfused recipients in Mexico

BACKGROUND: Transfusion-transmitted viral infection (TTI) is a major problem in patients receiving blood products. Monitoring high-risk patients is essential for assessing the epidemiology of blood-borne infections.
STUDY DESIGN AND METHODS: A 1-year, cross-sectional seroprevalence study in patients with a history of multiple transfusions was conducted. Peripheral blood samples were titered to detect serologic markers of human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV). The presence of these viruses and demographic, behavioral, and medical traits were assessed.
RESULTS: A total of 300 male and female multiply transfused patients with a mean age of 30.7 (±17.5) years were studied. The prevalence was 13.7% for HCV, 7% for HBV, and 1.7% for HIV. Patients with hemophilia had the highest prevalence for HCV and HIV infections, and hemodialyzed patients, for HBV infection. The risk factors related to acquired HCV were hemophilia (odds ratio [OR], 5.6; 95% confidence interval [CI], 2.5-12.6), more than five hospitalizations (OR, 3.8; 95% CI, 1.6-8.9), and having received a transfusion before mandatory screening in 1993 (OR, 8.4; 95% CI, 2.0-34.6), and for HIV, having received a transfusion before 1987 (OR, 19.0; 95% CI, 2.0-177.7). The main risk factors for HBV were having end-stage renal disease and being treated with hemodialysis (OR, 3.7; 95% CI, 1.4-9.9) and transplantation (OR, 4.2; 95% CI, 1.4-12.1).
CONCLUSIONS: This study showed that HCV infection was more frequently identified than HBV and HIV infections in multiply transfused Mexican patients. Additionally, several risk factors are associated with TTI such as mandatory screenings before 1987 and 1993, which were the most important for HIV and HCV infections but not for HBV.     

PEG沉降病毒富集法对极低载量HBV感染血浆的确认效果评价

目的评价PEG沉降病毒富集法对极低载量HBV感染血浆的确认效果。方法建立PEG沉降病毒富集法(简称PEG富集法)并确认其HBV病毒富集效率;以本地区优势C型HBV毒株确认HBV序列多区段巢式PCR和qPCR的检测灵敏度;比较PEG富集法和超高速离心法分别与HBV序列多区段扩增相结合(分别简称为PEG体系和超离体系)时,对核酸初检反应性而鉴别试验无反应性即非重复反应性(Non-repeatable Reactive,NRR)但含有极低病毒载量的标本进行HBV感染确认的效果差异。结果 PEG富集法在30 IU/mL浓度的病毒富集效率为(81.5±30.3)%;巢式PCR对BCP片段扩增的检测具有最佳灵敏度为1 IU/mL;PEG体系灵敏度的推测范围为(0.12-3.07)IU/mL。69份NRR标本经PEG体系和超离体系分别确认出HBV感染标本37份(53.6%)和46份(66.5%),2体系对HBV感染的总确认率无差异(P>0.05)。但配对比较显示,超离体系在BCP和PreC/C区段扩增确认率显著高于PEG体系(P<0.05)。结论本研究建立的PEG富集法的成本较低、简单易行,可以作为超高速离心法的替代方法应用于普通血筛实验室对核酸检测反应性血浆的HBV感染的确认,但需要结合多个不同HBV序列区段的扩增检测以提高极低病毒载量血浆的确认率。     

Mini-pool screening by nucleic acid testing for hepatitis B virus, hepatitis C virus, and HIV: preliminary results

BACKGROUND: The purpose of this study was to evaluate the feasibility of nucleic acid testing (NAT) of mini-pools as a blood donation screening test. STUDY DESIGN AND METHODS: The stepwise implementation of NAT of mini-pools began in January 1997. Since March 1997, all blood donations collected by the German Red Cross Blood Transfusion Service of Baden-Wurttemberg were tested for hepatitis B virus (HBV), hepatitis C virus (HCV), and HIV nucleic acids. An extra barcoded serum sample is collected from each blood donor for NAT-based screening, which is performed only on hepatitis B surface antigen-, anti-HCV-, anti-HIV-, and anti-Treponema pallidum-seronegative donations. Samples are pooled to a maximum of 96. Positive results are resolved through intersecting subpools (a chessboard design). NAT-based screening does not include a virus concentration step before nucleic acid extraction. RESULTS: By the end of October 1997, 331, 783 donations in 3,779 pools had been screened. As yet, no viremic but seronegative blood donor has been found for the three markers. CONCLUSION: It is feasible to incorporate NAT-based screening of mini-pools into the routine virus diagnostics of a large blood transfusion service. It remains to be determined whether screening blood donations by NAT will indeed increase the safety of blood supply.     

亚甲蓝光化学灭活对血浆丙型肝炎病毒基因组核酸降解的研究

目的研究丙型肝炎病毒(HCV)基因组核酸在亚甲蓝光化学法(methelene blue photochemistry,MB-P)灭活病毒前后的变化,在全基因组水平分析MB-P对基因组各结构区段的降解作用。方法含HCV的血浆中加入终浓度为1.0μmol/L的MB,经约30000Lux强度的荧光照射后,在不同作用时间点取样;将HCV全基因组序列分为互相重叠的8个区段,分别进行RT-PCR,分析基因组核酸的完整性;同时运用实时定量PCR(real time-PCR,RT-PCR)技术观察核酸降解的动力学变化。结果基因组各区段RT-PCR结果发现,经过不同的光照时间,HCV基因组各区段的稳定性不同,第2、4、5、6区段对MB-P作用较敏感,基因组5’端区段和3’端区段经MB-P作用后的稳定性高于基因组其它区段;RT-PCR结果显示,随着光照时间延长,可被检测到的病毒核酸拷贝数逐渐下降。结论MB-P灭活过程中HCV基因组核酸被降解,而且基因组不同区段对光化学作用的反应性不同,提示RNA降解可能是病毒灭活的重要机制;检测病毒核酸稳定性以监测病毒灭活具有一定临床实用价值。     

Incidence of hepatitis C virus RNA in anti-HCV negative plasma pools in Croatia.

The risks of transmitting viral infection by blood and products derived from plasma have long been known and still remain an area of concern. Blood banks and transfusion centres are faced with the imminent introduction of nucleic acid amplification testing (NAT) of plasma pools as used by the plasma industry. In this paper, we show a part of our results of a validation study of an in-house method for routine polymerase chain reaction (PCR) screening for hepatitis C virus (HCV) RNA in plasma pools and the results of testing 2,718 anti-HCV negative plasma pools for the presence of HCV RNA. The European Committee for Proprietary Medical Products (CPMP) recommended that from 1 July 1999, only batches derived from plasma pools tested and found non-reactive for HCV RNA, using validated test methods of suitable sensitivity and specificity, should be batch released by authorities. The quality and efficiency of NAT detection of HCV RNA is among others influenced by the efficacy of RNA isolation, the primer selection and the use of control samples. Using modern molecular biology techniques (sensitive and specific in-house amplification methods for detection of HCV RNA and automated sequencing), we analysed samples of plasma pools from different Croatian transfusion centres. By detection of HCV RNA in an NIBSC working reagent (genotype 3) and a Pelispy HCV RNA run control (genotype 1) we determined a high reproducibility and sensitivity (below 100 International Units (IU)/ml) for our in-house method. By direct sequencing PCR cDNAs we proved the specificity of the test system and the possibility of determining the HCV genotype when the method was used for PCR screening of HCV RNA in single donations. Of 2,718 anti-HCV negative plasma pools we have found that 2.1$ were HCV RNA positive. Results of our investigation confirm the necessity of testing HCV RNA in plasma pools to further increase the safety of human plasma-derived drugs.