核酸扩增技术(NAT)在血液HBV DNA筛查中的应用研究

输血相关传染病的预防和控制已成为全社会关注的焦点,随着检测技术的不断进步,输血传播相关病毒的危险性大大降低,目前酶免疫检测(ELTSA)技术已经广泛应用于血液筛查,但每年仍有少量新发输血后肝火病例报道,如美国无偿献血者每单位供血传播HBV的危险性为1/6.3万.这些危险主要来自病毒感染者"窗口期"献血、病毒变异、低病毒载量感染及人工操作失误等,其中"窗口期"漏检是影响血液安全性的主要问题.     

核酸扩增技术(NAT)在血液HBV DNA筛查中的应用研究

输血相关传染病的预防和控制已成为全社会关注的焦点,随着检测技术的不断进步,输血传播相关病毒的危险性大大降低,目前酶免疫检测(ELTSA)技术已经广泛应用于血液筛查,但每年仍有少量新发输血后肝火病例报道,如美国无偿献血者每单位供血传播HBV的危险性为1/6.3万.这些危险主要来自病毒感染者"窗口期"献血、病毒变异、低病毒载量感染及人工操作失误等,其中"窗口期"漏检是影响血液安全性的主要问题.核酸扩增技术(NAT)可直接检测病毒核酸,大大缩短病毒ELISA检测的窗口期及检出变异株,发达国家已陆续将其纳入献血者的常规筛查项目,并取得一定的成效.但由于进口 NAT试剂成本很高,取制了该技术在我国血液常规筛查中的推广应用.为了进一步提高血液的安全性,降低血液核酸筛查的成本,笔者尝试将国产NAT试剂应用到血液HBV DNA筛查中,并对在献血者HBV DNA检测中建立一套适合中国国情的筛查检测体系的可行性进行了探讨.     

成都地区无偿献血者中HIV感染的状况分析

目的：控制经输血途径传播HIV的工作十分重要，对无偿献血者中HIV感染状况的分析，利于从源头上有效阻断HIV的传播。方法：对2000—2003年成都地区无偿献血者290138人份的血液，按卫生部《献血者血液检查标准)，采用HIV-1／2混合型两种ELISA诊断试剂盒检测血液中HIV抗体，经复检HIV抗体仍为阳性的血液，再用免疫印迹(WB)法最后确认。结果：无偿献血者中14例被确认为HIV感染者。结论：成都地区献血者中HIV感染率逐年呈上升趋势，提示应加强预防经血液HIV的传播，尽快提高血液HIV的检测技术水平，增强医务人员自我保护意识，确保临床患者的输血安全。     

51528例壮族献血者HIV感染情况调查

目的了解百色市壮族献血者HIV感染情况以及评估现行献血者HIV筛查策略的效果,确保临床用血安全有效。方法回顾性调查百色市51 528例壮族献血者的HIV感染情况。HIV筛查全部使用ELISA(抗-HIV1/2)法,初、复检均使用国产试剂,初、复检阳性标本送广西壮族自治区疾病预防控制中心HIV确认实验室检验。结果共检测标本51 528例,初检阳性45例,复检阳性31例,经确认试验阳性24例。献血者HIV感染率为0.047%。结论严格血源管理、重视HIV筛查检测、引进血液病毒灭活技术、运行良好的质量管理体系及加强临床科学合理用血等措施可以减少经血液途径传播HIV概率。但阻断经输血传播AIDS最有效的方法是建立一支固定的自愿无偿献血者队伍。     

献血时是否需要检查HIV抗原?

确认有无HIV感染一般行抗体检查。但从感染到检测出抗体需要4～8周。从感染到可以检测到抗体的时期叫做窗口期(windowperiod)。据报道这个时期血液中的病毒含量较多。目前,献血者的筛查只做抗体检查。如果处于窗口期的献血人员献血就可能被漏诊。因此,美国FDA向血液中心提出,为提供可靠的血液应行p24抗原检查,并已于去年开始检查。日本参考美国FDA引入p24抗原检查的过程和实施后的阳性率,从HIV感     

加强血液病毒检测 提高输血安全性

输血是临床治疗的重要手段之一 ,但是输血治疗和其他许多临床治疗措施一样 ,输血除治疗病人外 ,也能引起一些不良反应和并发症 ,特别是输血存在传播病毒的危险。为了保证输血的安全 ,我站已按照有关规定 ,采取了一系列措施 ,其中加大了宣传的力度 ,让献血者了解献血知识 ,做到自我排除 ,以及血液中检测病毒标志物对保证输血安全起到了重要的作用。1　输血可能传播的病毒　人类免疫缺陷病毒(HIV) ,血液是HIV病毒传播的三个重要途径之一 ,据有关资料统计 ,全球HIV感染者约 5 %～10 %经血液传播而感染 ,因此这一途径成为控制和预防AIDS的…     

血清学检测联合核酸检测在血液中心的应用价值

目的探讨在血液中心应用血清学检测联合核酸检测的价值,以提高输血的安全性,降低输血感染风险。方法以我血液中心于2015年1月至2015年12月期间大约共计3000份献血者血液标本作为本组研究的观察对象,所有血液标本均经血清学检测包括HbsAg、HCV抗体、HIV抗原/抗体、梅毒特异性抗体(以下简称酶免),与谷丙转氨酶;并行HBV/HCV/HIV3联检核酸定性检测。对于单纯核酸检测反应性的标本需进行鉴别试验,其中鉴别阴性的献血者进行跟踪检测,并将HbsAg、HCV抗体、HIV抗原/抗体三项酶联免疫检测结果与核酸检测结果进行对比分析。结果本组大约3000份血液标本中,经酶联免疫检测单试剂不合格331份,不合格率为1.35%;其中经酶联免疫检测双试剂反应性且经核酸检测无反应性的不合格标本共31份,其中HbsAg反应性11份,HCV抗体反应性20份;单纯核酸检测反应性标本共43份,经跟踪检测确认其中HBV13例、HIV窗口期感染2例,末检出HCV献血者。结论核酸检测与血清学检测的结果存在一定差别,核酸检测虽然对血清学阴性的HBV感染检出效果明显,但对于感染HBV但病毒载量极低者,其漏检的可能性较高;需要经跟踪检测与实验室检测以进一步确认。     

15688例献血员抗-HIV抗-HCV HBsAg梅毒检测结果分析

血液途径是HIV、HCV、HBsAg、梅毒的主要传播途径之一,对献血者进行严格检测是保证血液质量和输血安全的关键.为了解我市HIV、HCV、HBsAg、梅毒在一般人群中的感染情况,确保输血安全,落实防治措施,我们将2000年至2001年在南充中心血站献血的15688名献血员抗-HIV、抗-HCV、HBsAg、梅毒的检测结果进行分析,现报告如下:     

核酸检测技术在长春地区血液筛查的初步应用

输血是感染HBV、HCV和HIV的主要途径之一,其直接原因是血液病毒检测漏检,而90%左右的血液检测漏检是由"窗口期"引起的。通过对血液进行输血相关病毒的NAT检测,可有效缩短窗口期,降低输血传播疾病的发生。     

运用核酸扩增技术对22326名献血者的检测情况分析

输血相关传染病的预防和控制已成为全球关注的焦点问题,如何有效地检测出病毒以及缩短窗口期,是当下血液筛查检测技术的发展趋势。由于国产酶联免疫吸附试验(ELISA)法检测试剂的灵敏度相对偏低,窗口期长,而核酸扩增检测技术(NAT)检测直接测定病毒RNA或DNA,可显著缩短窗口期,降低输血感染的风险,提高用血安全,已成为不少地区血站开展献血者血液筛查的必要手段。2010年我国部分血站开始试点核酸检测,2012版《血站技术操作规程》,将NAT纳入血液检测项目,开启了我国NAT检测时代。按照国务院《卫生事业发展“十二五”规划》等有关文件要求,全面开展血站筛查核酸检测(以下称核酸检测),缩短病毒检测“窗口期”,是进一步提高我国临床用血安全水平、降低经输血传播疾病风险的重要举措。笔者通过对我站2017年1至12月份NAT检测情况统计分析,探讨开展核酸检测对献血者血液筛查的重要性。     

Laboratory strategic defense initiatives against transmission of human immune deficiency virus in blood and blood products

Serological methods based on enzyme linked immunosorbent assay (ELISA) and Western blot tests for detecting the presence of antibodies against the human immune deficiency virus are the standard techniques for identifying infected blood donors. However, these tests could not detect infected seronegative donors who were in the window period at the time of donation. Such donors can be identified by more elaborate methods including antigen detecting ELISA and polymerase chain reaction, which can detect viral antigens and nucleic acids in infected donor blood even in window period. In addition, the process of donor selection whereby individuals who were at high risk for HIV infections were excluded from the donor panel had substantially reduced the risk of window period donation. Furthermore, in order to ensure greater safety, transfusion centers nowadays undertake additional measures in the form of virucidal techniques such as the use of heat, detergents and photochemical agents to treat blood and blood products. Despite all of these measures, a risk-free transfusion was not practically achievable. However, risk-free transfusion is now possible with the introduction of recombinant blood products, the use of which is severely limited by their cost. Nonetheless, a risk-free transfusion is still achievable at a relatively little cost by transfusing suitably eligible patients with their own blood through the autologous blood transfusion program. Antibody testing is virtually the only method currently available in Nigerian blood banks. There is the need to reactivate and expand the scope of our National Blood Transfusion Service in order to make our blood and products safer.     

42397名献血者抗-HCV、抗-HIV、抗-TP、HBsAg检测结果分析

目的提高献血者血液检测准确度,最大限度减少人为因素造成因输血感染病毒性疾病的发生。方法两种以上的试剂按规定时献血者血液进行双检,并对结果进行分类统计及分析。结果42397名献血者中HBsAg阳性192例,阳性率0．45％,结果不确定47例;抗-HCV阳性187例,阳性率0．44％,结果不确定124例;抗-HIV阳性8例,阳性率0．02％,结果不确定14例;抗-TP阳性319例,阳性率0．75％,不确定78例;4项检测共计阳性969例,阳性率2．29％。结论献血人群中有较高病毒携带率及梅毒螺旋体感染率,加上科学技术发展水平限制及人体感染病毒后窗口期的存在,使输血存在引起病毒性疾病传播的危险。血站系统在对献血者的筛查工作中应不断加强管理,优化试剂组合,最大限度杜绝输血感染疾病的发生。     

Seroprevalence of Hepatitis B, Hepatitis C, HIV Infections in Blood Donors of Khulna, Bangladesh

To determine changes (Trends) in infection rates of Hepatitis B surface antigen (HBsAg), Hepatitis C virus (HCV) and Human immunodeficiency virus (HIV) in blood donors of Khulna Population. Specimens of 34953 voluntary blood donors & party or relative donors in Transfusion Medicine Department of Khulna Medical College Hospital from 2007 to 2009 were screened for HBsAg, anti-HCV, anti-HIV 1 & 2 reactivity in a cross-sectional survey by rapid test method. Reactive samples were verified using a recognized confirmatory test which consisted of a second generation enzyme immune assay (HBsAg), anti-HCV antibodies by anti-HCV EIA & for HIV by western Blot, respectively. The seroprevalence of HBsAg, Anti-HCV, HIV antibody 1 & 2 was 1.4%, 0.09% & 0.03% respectively in all blood donors. Prevalence of confirmed positivity was 0.62% for HBsAg, 0.04 % for Anti-HCV, 0.02% for HIV Western Blot. Between 2007 to 2009 a decreasing trend was observed in HBsAg frequency, HCV frequency decreased in 2009 compared to 2007. One HIV positivity found in 2009. Although the frequency of transfusion transmitted infections is low, party or relative donors have some risk factors than voluntary blood donors. Through more scrutiny in donor selection, improved serological test & reevaluation of infections routes in donor, infection reduction can be achieved.     

Risk of HIV infection from blood transfusion in Montreal

OBJECTIVES: To determine the incidence (including associated donor characteristics and time trends) of HIV infection among repeat blood donors and to estimate the risk of HIV transmission from blood transfusion in Montreal and in Canada as a whole. DESIGN: Retrospective cohort analysis. SETTING: Montreal Centre Blood Transfusion Service. PARTICIPANTS: People who donated blood at least twice after Nov. 1, 1985, and at least once from Apr. 1, 1989, to Mar. 31, 1993. INTERVENTION: Blood was screened for HIV by enzyme-linked immunosorbent assay and results were confirmed by Western blot analysis. OUTCOME MEASURES: Incidence density (the incidence rate per person-time) of HIV infection among repeat blood donors by sex, age group and region of residence, and incidence density and risk among first-time donors and for Canada as whole. RESULTS: There were 200,196 eligible donors and 432,631 person-years (PY) of observation. From 1989 to 1993, there were 18 HIV seroconversions among repeat donors. The crude incidence density was 3.3 per 100,000 PY (95% confidence interval [CI] 1.8 to 5.4 per 100,000 PY); it was 4.9 per 100,000 PY among men and 0.61 per 100,000 PY among women. Age-specific incidence per 100,000 PY was 2.5 among those 12-29 years of age, 5.1 among those 30-49, 2.9 among those 40-49, and 1.4 among those 50 and older. Based on an estimated mean "window period" (from when a donor's blood is capable of transmitting HIV until detectable antibody appears) of 25 days, the current risk of HIV infection from repeat donors in the window period is estimated at 1 in 440,000. Inclusion of blood units from first-time donors produces an overall risk of 1 in 390,000 (95% CI 1 in 250,000 to 655,000). The estimated risk per blood unit in Canada as a whole is 1 in 913000 (95% CI 1 in 507,000 to 2,050,000). CONCLUSIONS: This "sentinel" population of repeat blood donors is subject to important trends in HIV spread. Therefore, estimating the incidence density of HIV infection in repeat donors provides insight into the epidemiologic characteristics of HIV infection at minimal expense. As a result of measures to improve blood safety, including HIV testing, the incidence of HIV infection among blood donors in Canada is low and the risk of HIV transmission from transfusion is extremely small, although not zero.     

南京地区TTV分子流行病学调查

目的 :探讨经血传播病毒 (TTV)在南京地区分子流行学状况及TTV可能的传播途径。方法 :选择各型病毒性肝炎 975例 ,献血员 30例 ,慢性肾功能衰竭长期血透者 1 50例及健康者 32例作为研究对象 ,用n -PCR法检测各种人群中 (TTVDNA)的感染情况。并研究TTVDNA阳性与输血的关系。结果 :在各种人群中TTVDNA的检出率分别为非甲～庚型肝炎 2 2 % ,病毒性肝炎 1 3 .9% ,献血员 33 .3 % ,长期血透者 2 7.3 % ,健康人 9.4% ,1 36例TTVDNA阳性病毒性肝炎患者仅 1 9.9% (2 7/ 1 36)有输血史。结论 :献血员及其它经常接触血制品的人群是TTV感染的高危人群 ,输血是TTV感染的途径之一 ,肠道传播可能也是TTV传播的一条途径     

商丘市艾滋病VCT门诊HIV检测结果分析

目的:了解2005年商丘市疾病预防控制中心艾滋病自愿咨询检测(HIV voluntary counseling&testing,VCT)门诊求询者HIV感染状况,为本市艾滋病的预防控制提供依据。方法:对2005年1月~12月在本中心艾滋病VCT门诊203例求询者血液标本进行了HIV_抗体(1/2)检测,用酶联免疫吸附(ELISA)试验初筛,初筛试验阳性者用蛋白印迹检测法(WB)作确认试验。结果:检出HIV_抗体阳性30例,阳性率14.78%;男15例、女15例,男女之比为1∶1。年龄分布在18~70岁之间,其中1995年前后有偿献血感染13例,1995年前后输血感染11例,阳性配偶感染2例,不洁性行为3例,接触者1例。职业构成以农民为主(86.67%)。结论:我中心艾滋病VCT门诊求询者HIV感染仍以1995年前后有偿献血者及受血者为主,应加强控制HIV感染从高危人群向普通人群传播、配偶间传播及母婴传播,尽快制定具体干预措施。     

Computer-based interview for screening blood donors for risk of HIV transmission.

OBJECTIVE--To test the ability of a computer-based interview to detect factors related to the risk of the human immunodeficiency virus (HIV) among potential blood donors and to determine donor reactions to the use of the interview. DESIGN--A comparison of the rate of detection of HIV-related factors elicited by a computer interview with that obtained by standard American Red Cross procedures for assessment of donor suitability, including a randomized crossover trial in which the order of the two methods was reversed. Information obtained by the computer was not available to influence the use of blood components for transfusion. SETTING--The computer interview was administered to donors at an American Red Cross blood donor center and at a mobile blood drive at a hospital. SUBJECTS--Consecutive sample of 294 male and female blood donors 18 to 75 years of age. MAIN OUTCOME MEASURES--Subjects' responses to the computer-based interview as well as responses to the standard Red Cross written questionnaires and face-to-face interviews were used for donor assessment. RESULTS--The interview took an average of 8 minutes to complete. From among 272 donors who provided complete data, the computer identified 12 donors who reported either behaviors associated with a risk of HIV acquisition or symptoms compatible with the acquired immunodeficiency syndrome; none of these donors had been so identified either by routine written questionnaires or by face-to-face interviews used to screen potential blood donors. Only one of the 12 identified donors used the confidential unit exclusion procedure to prevent use of his donated unit. The rate of elicitation of HIV-related factors by the computer interview was 12 (4.4%) of 272 (95% confidence interval [CI], 2.3% to 7.6%), compared with two (0.13%) of 1536 (95% confidence upper bound, 0.28%) using the standard Red Cross procedure (P less than .0001). Tests for antibodies to HIV were negative in blood samples from all of the 272 subjects studied. The subjects enjoyed the computer interview and judged it to be more private than the standard donor assessment method. They also predicted that donors would be more honest with the computer interview than with a human interviewer. CONCLUSIONS--Computer-based screening elicits more HIV-related factors in the health histories of blood donors than do the standard questionnaire and interviewing methods currently in use. Computer-based screening is also acceptable to blood donors.     

Transfusion Transmitted Infections in Armed Forces: Prevalence and Trends

Background

This study presents data on the prevalence rate of infectious markers among voluntary and replacement donors in the blood transfusion service in Armed Forces from 2000 to 2004.

Methods

39,646 units of blood were collected from donors during the period from 2000 to 2004. All the samples were screened for hepatitis B surface antigen (HBsAg), human immunodeficiency virus (HIV) 1&2, hepatitis C virus (HCV), and by venereal disease research laboratory test (VDRL).

Results

24,527 (61.9%) were voluntary donations and 15,119 (38.1%) replacement donations. Prevalence of HBsAg had decreased, amongst voluntary donors from 1.67% to 0.77% but the positivity rate has not showed significant change. Seropositivity of HIV had decreased both in voluntary and replacement donors to 0.22% and 0.86% respectively. The seropositivity for anti-HCV showed steady decrease amongst voluntary donors from 0.46% to 0.20% in 2004, but in replacement donors, there was an increase in reactivity rate from 0.43% to 0.65%.

Conclusion

The increased seropositivity for HCV, HIV and HBsAg could be decreased by introduction of nucleic acid amplification testing (NAT) in minipools for HCV and HIV and introduction of anti-HBcAg (IgM) for hepatitis B virus (HBV) infection. But this may not be possible in near future in developing countries due to financial constraints. At present implementation of strict donor criteria and with use of sensitive laboratory screening tests it is possible to reduce the incidence of transfusion transmitted infections (TTI) in Indian scenario.Key Words: Transfusion transmitted infections, Human immunodeficiency virus, Hepatitis C virus, Hepatitis B virus     

Rapid latex agglutination assay using recombinant envelope polypeptide for the detection of antibody to the HIV

Unscreened blood transfusions continue to be one of the major modes of transmission of the human immunodeficiency virus (HIV) in developing countries, such as in Central Africa, where 5% to 18% of blood donors are HIV seropositive. We evaluated a rapid latex agglutination assay using a novel recombinant envelope polypeptide of HIV for the detection of HIV antibodies among 2820 blood donors and clinical patients from diverse geographic regions, including on-site testing in Central Africa of 1600 blood donors. Overall, 29.2% of the serum samples were positive by Western blot assay. On a single determination, the latex agglutination slide test was found to be highly sensitive and specific compared with Western blot results in these populations with a relatively high prevalence of infection. Use of this assay will allow the immediate implementation of serologic screening for HIV in developing areas of the world, where standard screening procedures are impractical or not available, and in many other clinical settings, such as sexually transmitted diseases clinics and hospitals, where testing and counseling could be promptly implemented.     

基于HIV检测分析对无偿献血人群HIV筛查策略及归队的探讨

目的 对陕西省域内无偿献血者进行人类免疫缺陷病毒（human immunodeficiency virus,HIV）检测,分析省域内无偿献血人群HIV流行情况,以期为制定适宜的血液筛查策略和献血者归队方法提供数据与理论依据。方法 选定有区域代表性的陕西省西安市、延安市与安康市3家中心血站的无偿献血者血液样本进行2种四代酶联免疫吸附试验（enzyme-linked immunosorbent assay,ELISA）和病毒核酸检测（nucleic acid testing,NAT）HIV,将所有检测呈反应性的样本进行确证试验。结果 对290 341例无偿献血者进行HIV筛查,总计352例初检结果为不合格（含1例ELISA-/NAT+有反应性）,筛查反应率0.12%;所有样本送至属地市疾控中心检测,最终确证阳性11例（含核酸检测阳性1例）,确证阳性符合率3.79/10万。结论 陕西省域内无偿献血者中HIV阳性符合率低,提示目前无偿献血者HIV的血液筛查策略有待改进,同时也为献血者归队提供一定的理论支持。