广东中山地区献血人群人类嗜T淋巴细胞病毒感染状况调查

目的了解中山地区人类嗜T淋巴细胞病毒(HTLV)在无偿献血人群中的感染情况。方法对2016年3-12月40 874份在中山地区无偿献血的血液标本,采用酶联免疫吸附试验方法(ELISA)进行HTLV抗体筛查,筛查呈反应性的标本进行双孔复查,复查仍阳性的标本使用免疫化学发光法(CLIA)检测,检测阳性的标本使用免疫蛋白印迹法(WB)进行确证,确证阳性的视为感染。结果中山地区40 874份无偿献血者中HTLV抗体ELISA检测阳性21例,ELISA检测阳性率为0.05%,免疫化学发光法检测阳性5例,WB确证阳性1例,感染率为0.002 4%。结论为了保证输血安全,降低输血感染HTLV,有必要对中山地区的初次献血者进行HTLV的筛查。     

山东省58395例献血者中人类嗜T淋巴细胞病毒感染的检测

目的了解人类嗜T淋巴细胞病毒(HTLV)在山东省献血人群中的感染情况。方法应用酶联免疫吸附试验(ELISA)、蛋白印记(western blot,WB)法和核酸检测(nucleic acid testing,NAT)技术,对山东省献血者58 395份标本进行HTLV感染调查。结果在58 395例献血者中用ELISA法检出31例呈抗体阳性,用WB法进行抗体确证试验结果为阴性,NAT确证病毒阳性为2例,HTLV阳性率为0.003 4%。结论山东省献血人群中存在HTLV感染,输血安全存在隐患,核酸检测为WB检测的有效辅助方法。     

广州番禺南沙地区无偿献血人群HTLV感染情况调查

目的了解珠三角局部沿海地区献血人群人类T淋巴细胞白血病病毒(HTLV)的感染状况,为国家卫生行政部门今后制定地区性血液安全政策提供参考依据。方法对本站无偿献血标本,采用酶联免疫双抗原夹心方法进行初筛,初筛阳性的标本进行双孔复查,对复查阳性的标本进行化学发光法检测,化学发光法阳性标本进行蛋白印迹法(WB)和病毒核酸检测法(PCR)确证。结果 2016年3月—2019年3月共筛查92 047例无偿献血样本(未剔除重复献血者样本),发现酶免初筛阳性标本34例,其中化学发光阳性5例(不存在酶免及化学发光重复阳性病例),蛋白印迹法及病毒核酸检测均阴性。结论广州市番禺及南沙地区无偿献血人群是HTLV感染非流行区,综合考虑血液安全和经济成本效益,降低HTLV经血传播风险,血液可采用滤白处理。     

4种ELISA试剂筛查献血人群抗-HTLV结果评价

目的　对HTLV抗体ELISA诊断试剂盒进行初步评价。方法　 797份献血者标本用 1种国产双抗原夹心法试剂 ,1种国产间接法试剂和 2种进口间接法试剂同时进行检测 ,对ELISA阳性血清均用Westernblot(WB)进行确证。结果　 797名献血者四种试剂盒均能检出 4份抗 HTLV 1阳性标本 ,但三种间接法试剂均分别出现了 4～ 11份不等的假阳性 ,而国产双抗原夹心法试剂则未出现假阳性。结论　国产HTLV抗体ELISA诊断试剂盒已具备用于规模筛检能力。     

荧光定量PCR法检测HTLV核酸方法的建立

目的建立荧光定量PCR (FQ-PCR)法从血浆标本中检测HTLV核酸,用于献血者HTLV筛查。方法在沈阳军区总医院和安徽省血液中心总共收集献血者标本3 043份,用自配核酸提取试剂提取HTLV核酸,用FQ-PCR法检测HTLV核酸。同时用ELISA方法进行anti-HTLV检测,如果ELISA呈反应性,进一步用免疫印迹实验进行确证,如果免疫印迹实验结果为不确定,间隔4周后追踪献血者,以确定是否为真阳性,以验证FQ-PCR检测方法的可行性。结果对留取的献血者标本进行扩增,所有扩增结果均为阴性。用ELISA方法对这些标本进行HTLV抗体检测,共有48份标本ELISA检测呈反应性(包括灰区0.5=S/CO 1.0),对这48份标本做确证实验,有16份标本出现条带,但结果均为不确定。间隔4周后对这16位献血者进行追踪,共追踪到11位献血者,再次留取标本进行检测,结果均为阴性。分别选择7种不同拷贝数的假病毒核酸进行最低检出限的确定实验,最低检出限可以达到1.56 Copies/mL。结论本研究建立的FQ-PCR方法可以检测HTLV核酸,可以用于献血者的HTLV核酸筛查。     

安徽地区献血人群HTLV感染调查

目的了解安徽地区无偿献血人群人类T淋巴细胞病毒感染状况,为本地区临床用血提供合适的安全筛查策略。方法对安徽省合肥、蚌埠、芜湖、安庆、马鞍山5地区部分献血人群采用ELISA试剂筛查HTLV-Ⅰ/Ⅱ抗体,筛查阳性标本采用蛋白印迹法(Westem blot,WB)确证和核酸检测,同时对献血者进行追踪、检测。结果 5地区共抽检献血者标本100 840份,ELISA法筛查出阳性标本57份,确证结果"不确定"2份,其余全部为阴性;确证阳性率为0%;追踪到献血者8人,结果7人仍为原ELISA试剂筛查阳性,确证结果均无阳性。结论安徽地区为HTLV低流行地区,可以暂不开展献血者HTLV全面筛查。     

2016—2018年广州献血者HTLV流行概况及基因分型

目的系统地了解广州地区无偿献血人群中HTLV感染的流行情况。方法采用电化学发光法对2016年3月—2018年4月广州血液中心635 456名献血者中ELSIA初筛阳性的129例标本进行抗-HTLV筛检,对ELISA及电化学发光法检测均为阳性的标本经RT-PCR或WB法检测确证并通过巢式PCR扩增确定其HTLV分型。结果 ELISA及电化学发光法筛查-HTLV均为阳性标本33例;经RT-PCR或WB法检测确证为HTLV-1型标本9例,阳性率为1.4/100 000(9/635 456)。巢式PCR扩增出gp46及LTR序列的8例标本均属于HTLV-1A亚型,其中1例为日本人种亚群,其余均为世界人种亚群。结论广州属于HTLV低流行地区,无偿献血人群中仅存在HTLV-1的感染其分型主要为HTLV-1A中世界人种亚群及日本人种亚群;献血者的HTLV常规筛查可有效杜绝HTLV的经血传播,进一步提高血液安全。     

在安全输血中应用丙型肝炎病毒核心抗原检测技术的初步探索

本研究探讨用丙型肝炎病毒（HCV）核心抗原酶联免疫吸附技术（HCV-cAg ELISA）筛查献血员感染HCV的可行性.对我医院2003年1月-2005年12月间的8 677份献血者血清标本进行抗-HCV初检和复检.将仅初检阳性的15份血清标本和仅复检阳性的14份血清标本,分别再做HCV-cAg ELISA和HCV RT-PCR检测.结果表明：经HCV-cAg ELISA检测,29份仅初检（15份）和仅复检（14份）抗-HCV阳性血清标本中只有5份结果为阳性,阳性率为17.24%.经HCV RT-PCR检测,29份仅初检和仅复检抗-HCV阳性血清标本中同样也只有5份阳性结果,阳性率也为17.24%.结论：HCV-cAgELISA检测技术的敏感性与HCVRT-PCR技术类似,但成本明显降低,有可能作为HCV感染的辅助确证试验,或作为抗-HCV检测技术的更新换代技术用于安全输血中献血者血液的筛查.     

湘潭地区无偿献血人群中HTLV-Ⅰ／Ⅱ感染的血清学调查

【目的】了解湘潭地区无偿献血人群中人类嗜T淋巴细胞白血病病毒（HTLV）的感染情况。【方法】用酶联免疫吸附试验方法（ELISA）筛查2008年湘潭地区的无偿献血人群血清标本830份。初筛阳性者用双孔试验确认,仍为阳性者采用免疫印迹法（WB）进行确证。【结果】湘潭地区830份献血标本中初检有5例HTLV-Ⅰ／Ⅱ抗体阳性,经双孔复测,全部结果为阴性。【结论】本次调查的无偿献血人群中没有HTLV-Ⅰ／Ⅱ感染者,可为临床安全用血提供HTLV的流行病学资料。     

南宁市无偿献血人群HTLV检测结果分析

目的了解南宁市无偿献血人群人类嗜T细胞病毒(HTLV)的感染状况,评估HTLV对南宁市血液安全现状的影响。方法使用HTLV酶联免疫吸附试验试剂盒对南宁市61 804份无偿献血者血液标本进行HTLV-Ⅰ/Ⅱ抗体筛查,筛查阳性标本应用免疫印迹试验和实时荧光定量聚合酶链反应进行确证。结果 61 804份无偿献血者血液标本共有HTLV-Ⅰ/Ⅱ抗体阳性24份,初筛阳性率为3.88,其中4份确证阳性,3份不确定,HTLV阳性率为0.65。结论南宁市无偿献血人群中存在一定的HTLV感染,但目前处于较低水平,仍需继续监测。     

厦门市无偿献血者HTLV感染情况及基因亚型分析

目的 了解人类T淋巴细胞白血病病毒(HTLV)在厦门市无偿献血人群中的感染情况及基因亚型的特点.方法 采用ELSIA方法自2004年2月起对来夏门市中心血站献血的全部献血者131 823人做抗-HTLV-1/2筛检,对ELISA检测阳性的标本经巢式PCR、Taqman MGB探针及WB法检测确认并确定HTLV亚型.结果 在5年1个月的时间里,ELISA法初筛抗-HTLV阳性58例;经巢式PCR或Taqman MGB探针检测确认为HTLV-1阳性的标本24例,阳性率为0.02％(24/131 823).分析其中16例阳性标本的gp46序列发现其都属于HTLV-1A亚型,其中15株序列属于世界人种亚群,另外1株属于日本人种亚群.结论 厦门市无偿献血者中存在HTLV感染且感染者均为HTLV-1型,以世界人种亚群为主,并有少量的日本人种亚群;对献血者的HTLV筛查可有效杜绝经输血传播HTLV,进一步提高输血安全.     

北京地区献血人群人类巨细胞病毒及人类T淋巴细胞白血病病毒感染情况调查

目的 了解北京地区人类巨细胞病毒(HCMV)及人类T淋巴细胞白血病病毒(HTLV)在无偿献血人群中的感染情况。方法 北京地区2 010例无偿献血者血液标本采用ELISA方法筛查HCMV-IgG,HCMV-IgM,HTLV-Ⅰ/Ⅱ抗体。初检阳性标本进行双孔复检,两次检测结果均为阳性标本判定为ELISA结果阳性,HTLV阳性样品经巢式PCR检测核酸进行确认。结果 北京地区2 010例无偿献血者中HCMV-IgM和HCMV-IgG阳性率分别为2.19%和92.59%,ELISA法初筛抗-HTLV阳性1例,巢式PCR确认结果阴性。结果 通过χ²检验统计学分析显示,无偿献血者HCMV-IgG阳性率男性低于女性(χ²=5.88,P＜0.05),18～25岁年龄段的献血者HCMV-IgG和HCMV-IgM阳性率低于其他年龄段(χ²=16.51,21.52; 均P＜0.05),大学生HCMV-IgG阳性率低于其他职业献血者(χ²=14.20,P＜0.05),而献血者教育程度与HCMV-IgG和HCMV-IgM阳性率无关(P＞0.05)。结论在该次调查中,北京地区2 010例无偿献血者中未发现感染HTLV的病例,而HCMV感染率与性别、年龄、职业相关,与教育程度无关。     

Strategies for diagnosis of HTLV-I and -II

BACKGROUND: No gold standard exists for diagnosis of HTLV infection. The aim of thus study was to compare the accuracy of a combination of two sensitive ELISAs with Western blot (WB), a line immunoassay, and PCR for diagnosis of HTLV infection. STUDY DESIGN AND METHODS: Nine hundred eighty-five specimens were tested for the presence of HTLV antibodies by HTLV-I and/or HTLV-II EIAs (Murex and Ortho), WB (Diagnostic Biotechnology), line immunoassay (INNO-LIA, Innogenetics), and/or presence of HTLV DNA by PCR. The results were compared with the probable HTLV infection status of each subject, as determined by detailed review of all available laboratory, clinical, and epidemiologic data. RESULTS: The sensitivity for diagnosis of HTLV-I infection was high for all assays evaluated, but both PCR and WB had a lower sensitivity rate (approx., 80%) for confirmation of HTLV-II. INNO-LIA detected 94 percent of the HTLV-II-positive samples. However, Murex EIA in combination with Ortho EIA was 100-percent sensitive for the detection of both HTLV-I and HTLV-II antibodies. Furthermore, the number of samples giving indeterminate results in the ELISA combination was much lower as compared with WB (2.5% vs. 50%). CONCLUSION: Based on these findings, a new, more sensitive and specific test strategy for HTLV diagnosis than the current algorithm, which includes WB, is proposed. Thereby, both the direct and indirect costs can be substantially reduced.     

Serologic and molecular typing of human T‐lymphotropic virus among blood donors in Maputo City,Mozambique

BACKGROUND: Screening for human T‐lymphotropic virus‐1/2 (HTLV‐1/2) infection is not performed in blood banks in Mozambique. The aim was to determine the prevalence of HTLV‐1/2 among blood donors of the Maputo Central Hospital Blood Bank and measure the coinfection rate of HTLV‐1/2 with human immunodeficiency virus (HIV), hepatitis B virus (HBV), and syphilis. STUDY DESIGN AND METHODS: A total of 2019 consecutive blood donors were screened for HTLV‐1/2 antibodies, HIV‐1/2 antibodies, hepatitis B surface antigen (HBsAg), and rapid plasma reagin (RPR) for syphilis. Specimens reactive on a first HTLV‐1/2 enzyme immunoassay (EIA) were retested using a second EIA. Specimens that were dually reactive on both EIAs were further tested using Western blot (WB) and real‐time polymerase chain reaction (PCR). RESULTS: All 18 dually reactive specimens (0.89%; 95% confidence interval, 0.48%‐1.30%) were positive for the presence of HTLV‐1 by WB and real‐time PCR. HTLV‐2 was not detected. The prevalences of anti‐HIV, HBsAg, and reactivity in the RPR test were 5.72, 6.01, and 0.98 percent, respectively. There was no significant association between HTLV‐1 infection and demographic variables (age and sex) or serologic markers (HIV, HBsAg, and RPR). For the 17 HTLV‐1–positive donors for whom serologic data for HIV, HBsAg, and syphilis RPR were available, 2 showed coinfection with HIV and 1 with HBV. CONCLUSION: Compared to other infectious agents, HTLV‐1 is present at relatively low levels among blood donors in Mozambique. Cost and logistics will present as major challenges for introducing HTLV‐1/2 screening in blood banks. In blood banks in Southern Africa where EIA testing is possible, a sequential algorithm of two EIAs may be a cost‐efficient option for HTLV‐1/2 screening.     

Sensitivity of Two Enzyme-linked Immunosorbent Assay Tests in Relation to Western Blot in Detecting Human T-Cell Lymphotropic Virus Types I and II Infection among HIV-1 Infected Patients from São Paulo,Brazil

We investigated the presence of human T-cell lymphotropic virus types I and II (HTLV-I and HTLV-II) infections, first searching for specific antibodies in 553 serum samples obtained from HIV-1-infected patients from São Paulo, Brazil. Sera were screened using two enzyme-linked immunosorbent assays (ELISAs): the ELISA-EM (ELISA HTLV-I/II, EMBRABIO, BR), which contains HTLV-I and HTLV-II lysates, and the ELISA-DB [ELISA HTLV-I/II, Diagnostic Biotechnology (DB), Singapore], which contains HTLV-I lysate, and HTLV-I and HTLV-II recombinant env proteins (MTA-1 and K55, respectively). Serum samples showing two positive and/or borderline results were confirmed by Western blot (WB 2.3, DB), which discriminates HTLV-I from HTLV-II. WB analyses disclosed 22 cases (4.0%) of HTLV-I and 34 (6.1%) of HTLV-II seroreactivity; 24 sera had indeterminate antibody profile (4.3%) and 2 specimens showed reactivity to both MTA-1 and K55 env proteins. Using stringent WB criteria and analyzing the population according to risk factors, the prevalence rates of HTLV-I and HTLV-II infections were 11.2% and 16.8% in IV drug users, 3.4% and 5.5% in heterosexual individuals, and 1.4% and 2.2% in homosexual/bisexual men, respectively. A comparison of ELISA and WB results disclosed that both ELISAs were highly sensitive in detecting HTLV-I antibodies, whereas the ELISA-DB showed 82% sensitivity and the ELISA-EM 100% sensitivity in detecting HTLV-II antibodies. PCR analyses conducted on 37 representative cells samples confirmed the presence of HTLV proviral DNA in the majority of concordant serological cases, except in one, which was HTLV-I infected and seroreacted with K55 protein of HTLV-II. Indeed, after PCR, one case of HTLV-I infection and HTLV-II coinfection, and 30% of WB-seroindeterminate or inconclusive cases infected with HTLV-II could be detected. Our data stress high prevalences of both HTLV-I and HTLV-II infections in HIV-1 coinfected i.v. drug users from São Paulo, and suggests that ELISA kits containing only K55 protein as the HTLV-II-specific antigen, may not have the appropriate sensitivity for the detection of HTLV-II infection in this geographic region, pointing out the need of improved screening tests to be used in Brazil.     

广东籍献血人群HTLV感染状况调查

目的了解东南沿海省份的元症状献血人群人类嗜T淋巴细胞病毒（human T cell lymphotvopic virus,HTLV）的血清学流行状况。方法从符合卫生部健康标准并经血液常规检测合格的标本中,依据籍贯选择性收集2500份血样,采用双抗原夹心的酶免吸附（EIA）方法进行HTLV-Ⅰ／Ⅱ抗体筛查。EIA初筛阳性结果经复检后,由蛋白印迹法（Western Blot,WB）进行确认试验。结果初筛获得EIA反应性样本5例,HTLV抗体阳性率为0．20％（5／2500）,均为广东籍。经蛋白印迹试验,仅1例OD值为3．00的EIA强反应者获确认为HTLV—Ⅰ型病毒感染,2例未获确认,2例为确认阴性。经追溯,确认HTLV—Ⅰ阳性者为无症状定期献血者,已有6次合格献血经历。结论尽管深圳献血人群HTLV流行率的总体水平较低,但多次献血的经历使经血传播HTLV的风险增大数倍。     

Low incidence of HTLV infections in random blood donors with indeterminate western blot patterns

Peripheral blood mononuclear cells (PBMCs) were recovered from platelet units of 61 blood donors who were HTLV-I positive and 3 blood donors who were HTLV-I negative on enzyme-linked immunosorbent assay (ELISA). Western blot analyses were performed on the sera and DNA was prepared from the PBMCs and analyzed by the polymerase chain reaction (PCR). Of the 61 repeatably reactive samples, 2 were positive, 26 were negative, and 33 were interpreted as indeterminate on Western blot. HTLV-II sequences were detected by PCR in one of the Western blot-positive samples, as well as in one Western blot-indeterminate sample that showed reactivity to p24 only. HTLV-I sequences were detected in the second Western blot-positive sample. HTLV sequences were not detected in the remaining samples, which suggested that the majority of individuals with indeterminate results on Western blots that used one set of commercially available reagents are not infected with HTLV. It is demonstrated in this study that PCR can be used not only to resolve the infection status of individuals with indeterminate Western blots but also to distinguish between HTLV-I and HTLV-II.     

Asymptomatic infection by Leishmania infantum in blood donors from the Balearic Islands (Spain)

BACKGROUND: Visceral leishmaniasis (VL) caused by Leishmania infantum is a zoonotic disease endemic throughout the Mediterranean basin. The existence of asymptomatic human infection entails the risk of transmission by blood transfusion. STUDY DESIGN AND METHODS: The prevalence of Leishmania infection was studied in 1437 blood donors from the Balearic Islands (Majorca, Formentera, and Minorca) using immunologic (Western blot [WB] and delayed-type hypersensitivity [DTH]), parasitologic (culture), and molecular (nested polymerase chain reaction [PCR]) methods. In addition, the efficiency of leukoreduction by filtration to remove the parasite was tested by nested PCR in the red blood cell (RBC) units. RESULTS: Leishmania antibodies were detected in 44 of the 1437 blood donors tested (3.1%). A sample of 304 donors from Majorca was selected at random. L. infantum DNA was amplified in peripheral blood mononuclear cells (PBMNCs) in 18 of the 304 (5.9%), and cultures were positive in 2 of the 304 (0.6%). DTH was performed on 73 of the 304 donors and was positive for 8 of them (11%). Of the 18 donors with positive L. infantum nested PCR, only 2 were seropositive. All the RBC samples tested (13 of 18) from donors with a positive PBMNC nested PCR yielded negative nested PCR results after leukodepletion. CONCLUSIONS: Cryptic Leishmania infection is highly prevalent in blood donors from the Balearic Islands. DTH and L. infantum nested PCR appear to be more sensitive to detect asymptomatic infection than the serology. The use of leukodepletion filters appears to remove parasites from RBC units efficiently.