中国厦门地区献血者细小病毒B19感染情况研究

目的:了解中国厦门地区献血者细小病毒B19感染情况。方法:对中国厦门地区部分献血者标本进行细小病毒B19核酸检测和抗体检测,对核酸检测阳性标本进行序列测定和基因型分析。结果:在总共10 452人份献血者标本中检出6例B19核酸阳性,阳性率0.06%,阳性标本的DNA定量结果为3.59×10~2-1.07×10~4IU/ml;6例核酸阳性标本测序分析结果均为基因I型。B19-Ig M抗体阳性率为4.64%(50/1078),B19-Ig G阳性率16.79%(181/1078);B19-Ig G阳性率随年龄增加而升高(χ~2=7.964,P0.05),与性别差异无关。结论:中国厦门地区献血者细小病毒的总体感染率较其他地区偏低,但也存在一定比例的病毒血症,在今后的输血保障工作中应引起重视。     

佛山地区无偿献血人群人细小病毒B19感染情况分析

目的了解佛山地区无偿献血人群人细小病毒(HPV)B19感染现状。方法采用ELISA检测血液中的HPV B19IgG和IgM抗体,并用PCR检测抗体阳性标本HPV B19DNA。结果 368例无偿献血者标本中检出HPV B19IgG阳性92例,阳性率为25.00%;检出HPV B19IgM阳性2例,阳性率为0.54%,两者比较差异有统计学意义(P0.01)。94例抗体阳性标本中,检测HPV B19DNA阳性4例,阳性率为4.26%。结论佛山地区无偿献血人群存在较高的HPV B19既往感染率,急、慢性感染率较低,慢性感染者HPV B19病毒载量较低。     

广东地区育龄妇女人细小病毒B19感染筛查结果分析

目的 通过对广东地区育龄妇女人细小病毒B19(human parvovirus B19,B19)感染筛查结果进行分析,了解广东地区育龄妇女人细小病毒B19感染情况. 方法 采用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测广东地区育龄妇女血清中人细小病毒B19特异性IgM和IgG抗体,并对检测结果进行分析. 结果 广东地区育龄妇女血清中人细小病毒B19特异性IgM和IgG抗体阳性率分别为1.40％(700/50 086)、16.72％ (341/2 039).结论 开展育龄妇女人细小病毒B19感染筛查工作,对优生优育、提高人口素质具有非常重要意义.     

快速诊断人细小病毒B19感染方法学比较研究

目的 比较本实验室自行构建的ELISA法与德国ELISA试剂盒方法及PCR法快速诊断人细小病毒B19感染的临床应用价值.方法 血液标本215份,分别采用B19病毒VP1独特区(VP1u)蛋白包被ELISA板建立B19病毒抗体检测方法(自行构建ELISA法),德国ELISA试剂盒方法检测标本B19病毒IgM与IgG,并采用PCR法检测B19病毒DNA,分析自行构建E1ISA法检测B19病毒的敏感性、特异性及准确度.结果 自行构建的B19病毒ELISA体系最佳抗原包被量为25 ng/孔,最佳标本血清稀释倍数为1∶ 200;以德国B19病毒ELISA试剂盒为对照,自行构建ELISA法检测B19病毒IgM的敏感性、特异性及准确度分别为79.07％,87.18％,94.89％,检测B19病毒IgG的敏感性、特异性及准确度分别为88.46％,90.79％,93.53％;以PCR法检测B19病毒DNA为标准,自行构建ELISA法检测B19病毒IgG的敏感性、特异性及准确度分别为90.32％,77.77％,95.71％.结论 本实验室自行构建的ELISA检测方法与德国B19病毒抗体检测ELISA试剂盒法及PCR法检测B19病毒DNA有较好一致性,可作为B19病毒感染常规诊断方法.     

人细小病毒B19抗体及抗心磷脂抗体(aCL)在流产人群中的检测价值

目的探讨人细小病毒B19抗体及抗心磷脂抗体(aCL)在流产人群中的检测价值,为流产的预防提供理论依据。方法本研究观察对象为2015年1月-2017年1月于我院妇产科就诊的386例自然流产患者以及年龄配对的200例自愿终止妊娠的孕产妇,分别设为观察组与对照组。抽取空腹肘静脉血4ml,离心分析血清后采用酶联免疫吸附试验(ELISA)检测两组孕产妇血清aCLIgG、aCLIgM水平和B19IgG、B19IgM水平,比较两组孕产妇B19IgG、B19IgM、aCLIgG、aCLIgM阳性率以及血清平均水平。结果观察组B19IgG、B19IgM、aCLIgG、aCLIgM阳性率分别为5.08%、41.71%、52.33%、55.44%,对照组分别为6.00%、11.00%、7.00%、7.00%,观察组均显著高于对照组,差异具有统计学意义(P0.05);观察组血清B19IgG、B19IgM、aCLIgG、aCLIgM水平分别为(1.65±0.23)U/mL、(1.76±0.27)U/mL、(4.54±0.23)U/mL、(22.43±3.32)U/mL,对照组分别为(0.67±0.12)U/mL、(0.54±0.15)U/mL、(2.34±0.15)U/mL、(6.51±1.02)U/mL,观察组均显著高于对照组,差异具有统计学意义(P0.05)。结论自然流产孕产妇B19抗体、aCL抗体水平均显著高于正常孕产妇,提示其可能参与孕产妇流产的发生,动态监测B19抗体、aCL抗体水平对自然流产的预防具有十分重要的意义。     

人细小病毒B19IgM抗体间接ELISA检测方法的初步建立

目的：利用原核表达并纯化的人细小病毒B19（HPV B19）结构蛋白VP1初步建立HPV B19 IgM酶联免疫吸附试验（ELISA）的检测方法。方法以纯化的重组蛋白包被酶标板,建立IgM 间接ELISA检测方法,确定Cut-off值,并进行初步的应用。结果建立间接 ELISA方法Cut-off值为0．25,灵敏度为84．60％,特异性为99．70％,与对照试剂盒检测结果符合率为99．47％;用自产试剂盒检测115份孕妇和1700份健康献血志愿者血清中B19 IgM抗体,阳性率分别为12．17％和1．59％。结论通过包被 HPV B19-VP1蛋白建立的间接ELISA检测方法可用于 HPV B19早期感染或急性感染的辅助诊断。     

人细小病毒B19与特发性血小板减少性紫癜

目的:探讨人细小病毒B19感染与成人特发性血小板减少性紫癜发病的关系.方法:采用酶联免疫吸附法对50例成人特发性血小板减少性紫癜患者和30例健康成人的血清标本进行人细小病毒B19-IgM,IgG及血小板相关抗体检测.结果:50例特发性血小板减少性紫癜惠者血清中人细小病毒B19抗体总阳性率56%(28/50),30例健康成人3例人细小病毒B19-IgG为阳性(10%),2组间差异有统计学意义(IgG组P<0.01,IgM组P<0.05).病毒感染阳性与阴性患者的血小板相关抗体差异无统计学意义(P>0.05).结论:成人特发性血小板减少性紫癜患者人细小病毒B19抗体总阳性率高,以IgG抗体为主,符合成人以慢性型特发性血小板减少性紫癜为主的特点.成人中人细小病毒B19感染与血小板相关抗体无明显相关性,提示成人特发性血小板减少性紫癜发病因素的复杂性.     

亚甲蓝病毒灭活血浆中人细小病毒B19抗原、抗体的检测与分析

目的检测亚甲蓝病毒灭活血浆中人细小病毒(human parvovirus,HPV)B19抗原和抗体,评估临床输注的安全性。方法随机抽取2016年2月5日~4月30日制备的亚甲蓝病毒灭活血浆1 056袋,采用ELISA法检测HPV B19抗原和抗体;将HPV B19抗体阳性血浆按献血者性别、年龄、血型进行分组统计分析。结果 1 056袋亚甲蓝病毒灭活血浆HPV B19抗原均为阴性,Ig G和Ig M抗体总阳性率为20.5%(216/1 056)、Ig G抗体阳性率为19.5%(206/1 056)、Ig M抗体阳性率为4.9%(52/1 056),不同来源献血者制备的血浆HPV B19抗体阳性率有差异,随年龄增加抗体阳性率呈上升趋势,献血者性别差异无统计学意义,A型和AB型抗体阳性率较高,B型和O型抗体阳性率较低。结论亚甲蓝病毒灭活血浆中HPV B19抗原、抗体检测结果显示,临床输注血浆感染HPV B19的风险不高,但对于免疫缺陷等特殊受血者应引起重视,建议进行输血前HPV B19检测。     

中国新疆部分地区蚊传黄病毒感染调查

目的 了解新疆部分地区不明原因发热患者蚊传黄病毒的感染状况。 方法 采用ELISA方法对新疆维吾尔自治区南部喀什地区伽师县和麦盖提县、北部伊犁地区察布查尔县不明原因发热人群血清标本进行流行性乙型脑炎(乙脑)病毒(Japanese encephalitis virus, JEV)、登革病毒(Dengue virus, DENV)、西尼罗病毒(West Nile virus, WNV) IgM抗体检测。对检测结果阳性的血清标本平行进行以上3种病毒的交叉中和试验。 结果 新疆南北部地区641例不明原因发热患者急性期血清中,乙脑病毒IgM抗体阳性率0.62%(4/641), 登革病毒IgM抗体阳性率2.96%(19/641),西尼罗病毒IgM抗体阳性率1.72%(11/641)。ELISA检测阳性患者血清中有6例存在乙脑病毒、登革病毒、西尼罗病毒中和抗体,滴度介于1 ∶ 10~1 ∶ 40之间。 结论 新疆发热患者急性期血清标本可以检测到乙脑病毒、登革病毒、西尼罗病毒IgM抗体,部分病例血清中存在中和抗体。     

三种方法检测梅毒螺旋体抗体低值标本的比较分析

目的探讨微粒子化学发光法(CMIA)、酶联免疫吸附试验(ELISA)及梅毒螺旋体抗体明胶颗粒凝集试验(TPPA)对梅毒螺旋体抗体低值标本检测的相关性分析及应用价值,为临床对梅毒的诊治提供更加准确的实验室依据。方法采用CMIA法及ELISA法对患者血清进行梅毒螺旋体抗体检测,收集低值血清标本(S/CO值1~8)525例,再用TPPA法进行检测。根据S/CO值将标本分为3组:A组(s/co1.00~2.99);B组(s/co3~5.99);C组(S/CO 6.00~8.00),比较各组CMIA法、ELISA法与TPPA法的符合率。结果 A~C组CMIA法与TPPA符合率分别为30.3%、68.6%、96.6%,ELISA法与TPPA法符合率分别为10.6%、31.9%、72.6%。两种方法的符合率差异均有统计学意义(P0.05)。结论对于CMIA及ELISA方法检测梅毒抗体测定值(S/CO)低值样本时,尤其是CMIA法测定值(S/CO)为1~5.99的样本,ELISA法测定值(S/CO)为1~7.99的样本,应与TPPA方法联合检测复检验证尤为必要,以减少假阳性,提高检测结果的准确性。且两种方法与TPPA法比较,CMIA法符合率高于ELISA法。     

Clinical manifestations and outcomes of parvovirus B19 infection during pregnancy in Japan

Parvovirus B19 is a small DNA virus. Infection with parvovirus B19 during pregnancy may cause serious complications in the fetus, including hydrops fetalis and fetal death. The purpose of the present study is to clarify the clinical manifestations and outcomes of parvovirus B19 infection during pregnancy. This prospective study enrolled 478 women with suspected B19 infections during pregnancy between 1999 and 2004. One hundred cases (21%) of B19 infection were detected in 478 pregnant women who had been exposed to B19. Serological infection was confirmed by measurement of B19-specific IgM and IgG in sera. Forty-nine cases reported maternal clinical symptoms and 51 cases were asymptomatic. Facial rash was the most common symptom, with 51% (25/49) of the symptomatic patients complaining of either a facial, body or limb rash. The most common infectious source was children living in the home. Overall, the incidence of adverse fetal effects (including hydrops fetalis and fetal death) related to intrauterine B19 infection was 7% (7/100), and all seven cases were exposed to B19 infection before 20 weeks of gestation. Although half of the cases with parvovirus B19 infections during pregnancy were asymptomatic, patients with adverse fetal effects tended to be symptomatic including rash and fever. These clinical data may supply useful information to produce clinical guidelines for managing B19 infection during pregnancy.     

Clinical illness due to parvovirus B19 infection after infusion of solvent/detergent-treated pooled plasma

BACKGROUND: Lipid-enveloped viruses such as HIV, HBV, and HCV can be inactivated by treatment with solvents and detergents. HAV and human parvovirus B19 lack lipid envelopes and are not inactivated. Solvent/detergent-treated pooled plasma (S/D plasma) contains neutralizing antibodies, but it is not known whether the parvovirus B19 antibody content is sufficient to prevent transmission of the disease. A patient is described who developed a clinical illness due to parvovirus B19 infection after the infusion of S/D plasma. CASE REPORT: A 36-year-old woman with myasthenia gravis underwent five plasma exchange procedures from January 15 to January 25, 1999, using albumin, except for 5 units of SD plasma given because of a low fibrinogen level. Four of the 5 units were implicated in a recall after high levels of parvovirus B19 DNA were found in several lots. Two weeks after the infusion, the patient developed fatigue, a rash, and severe polyarthralgias. Parvovirus B19 IgG and IgM antibody titers were consistent with an acute infection. CONCLUSION: Clinically apparent parvovirus B19 infection can follow the use of S/D plasma that contains high levels of parvovirus B19 DNA.     

Diagnosis of parvovirus B19 infection in hematological patients with partial red cell aplasia of the bone marrow

AIM: To assess diagnosis of parvovirus B19 infection (PI) in patients with aplastic crises by combined use of polymerase chain reaction (PCR) and enzyme immunoassay (EIA) of specific IgM and IgG. MATERIAL AND METHODS: A total of 159 serum samples from 77 PI suspects were examined. The examination for virus DNA was conducted with modified "net" PCR in 108 samples, for specific IgM and IgG with EIA in 110 samples. RESULTS: The percentage of patients infected with parvovirus detected by PCR or EIA reached 60%. 21 of 77 patients with hemolytic anemias were infected with parvovirus B19, the virus persisting in 8 cases (40%). persistence of the virus was registered if viremia occurred in immunodeficiency due to the disease or immunosuppressive therapy. Immunity to parvovirus has not developed: IgM expression was the same as in patients without hemopoietic abnormalities, while IgG was not detected. The absence of specific immunity to parvovirus B19 occurred in patients treated with immunosuppressive drugs early after the end of viremia period in high IgM level and at the initial phase of IgG synthesis. IgM levels also remained unchanged; the level of IgG declined and was not identified furthermore. There were cases of reinfection. CONCLUSION: Combined use of PCR and EIA is optimal for diagnosis of parvovirus B19 infection in patients with hemolytic anemias. It was found that there are correlations between defects in specific immunity, persistence and immunodeficiency onset regarding viremia. Abnormal for the disease course levels of IgM and IgG indicate the persisting virus, the condition of specific immune response to parvovirus B19 and feasibility of reinfection. Reliable diagnosis of parvovirus infection is possible only in simultaneous use of PCR and EIA.     

Laboratory diagnosis of parvovirus B19 infection.

The sensitivity and application of the polymerase chain reaction (PCR) for the diagnosis of parvovirus B19 (B19) infection was investigated by simultaneously assaying a collection of 279 consecutively received samples for presence of anti-B19 IgM and IgG antibodies by Western blot and for B19 DNA by PCR and dot-blot hybridization (dot-blot); samples were sera from patients with suspected B19 infection. PCR and dot-blot detected B19 DNA in 9% (16/179) and 1% (2/179), respectively of Ab-positive samples (IgM+/IgG-, IgM+IgG+, IgM-IgG+), and in 28% (15/54) and 2% (1/54), respectively, of IgM+ samples. PCR also detected B19 DNA in 2% (2/100) of IgM-/IgG- samples, both of which had normal total IgG and IgM levels. PCR is of unique value because it permits diagnosis of B19 infection even in the absence of specific acute phase (IgM) and in the presence or absence of convalescent-phase (IgG) Ab.     

Laboratory diagnosis of cytomegaloviral infection in patients with anemia and hemoblastosis in the Omsk region

Research was carried out in 335 blood specimens of patients in the age of 3-35 y.o. in order to optimize diagnosis and treatment of such patients with aplastic anemia and hemoblastosis who got hemotransfunction to eliminate cytomegaloviral infection (CMVI). IgM were found out in 37.9% cases (2.8 times higher than in donors), low-avide IgG--in 44.8%. "early" proteins CMV--29.9% and DNA--in 36.8% cases. Concerning the DNA presence, preference was given to research of leucocytic suspension compared with blood serum. Of 28 children of 3-13 y.o. with anemia being seropositive in CMV, IgG antibodies were detected in 13 children while IgM antibodies to Parvovirus B19 were found in 10 children. 7 children with a grave form of disease showed combined infection of Parvovirus B19 and CMV with activation signs. It is not excluded that parallel influence of Parvovirus B19 on erythrocytic hemopoiesis growth and that of CMV on lymphocytic-monocytis cells aggravates immunodeficiency and promotes development of infection complications.     

Quantification of parvovirus B19 DNA using COBAS AmpliPrep automated sample preparation and LightCycler real-time PCR

The COBAS AmpliPrep instrument (Roche Diagnostics GmbH, D-68305 Mannheim, Germany) automates the entire sample preparation process of nucleic acid isolation from serum or plasma for polymerase chain reaction analysis. We report the analytical performance of the LightCycler Parvovirus B19 Quantification Kit (Roche Diagnostics) using nucleic acids isolated with the COBAS AmpliPrep instrument. Nucleic acids were extracted using the Total Nucleic Acid Isolation Kit (Roche Diagnostics) and amplified with the LightCycler Parvovirus B19 Quantification Kit. The kit combination processes 72 samples per 8-hour shift. The lower detection limit is 234 IU/ml at a 95% hit-rate, linear range approximately 10⁴-10¹⁰ IU/ml, and overall precision 16 to 40%. Relative sensitivity and specificity in routine samples from pregnant women are 100% and 93%, respectively. Identification of a persistent parvovirus B19-infected individual by the polymerase chain reaction among 51 anti-parvovirus B19 IgM-negative samples underlines the importance of additional nucleic acid testing in pregnancy and its superiority to serology in identifying the risk of parvovirus B19 transmission via blood or blood products. Combination of the Total Nucleic Acid Isolation Kit on the COBAS AmpliPrep instrument with the LightCycler Parvovirus B19 Quantification Kit provides a reliable and time-saving tool for sensitive and accurate detection of parvovirus B19 DNA.     

Hepatic dysfunction due to parvovirus B19 infection

Parvovirus B19 has been associated with different diseases. However, hepatic involvement has rarely been reported in adult patients with parvovirus B19 infection. Herein, we report two adult patients with hepatic dysfunction associated with acute parvovirus B19 infection. We believe that hepatic involvement in the setting of parvovirus B19 infection is more common than suspected, and that antibodies against this agent should be routinely measured in patients with hepatic dysfunction of unclear etiology. Received: August 1, 1999 / Accepted: November 29, 1999     

Prevalence and quantitation of parvovirus B19 DNA levels in blood donors with a sensitive polymerase chain reaction screening assay

BACKGROUND: Blood donor parvovirus B19 DNA prevalence with sensitive nucleic acid test assays has recently been demonstrated to be higher than that found with assays designed to detect high viral titers in the plasma manufacturing sector. STUDY DESIGN AND METHODS: Stored plasma aliquots from 5020 donations collected between 2000 and 2003 at seven US blood centers were tested. Testing was performed with a real-time B19 DNA polymerase chain reaction (PCR; TaqMan, Applied Biosystems) assay with a 50 percent limit of detection (LOD) of 1.6 IU per mL (95% confidence interval [CI], 1.2-2.1 IU/mL) and a 95 percent LOD of 16.5 IU per mL (95% CI, 10.6-33.9 IU/mL). Confirmation and quantitation of B19 DNA was accomplished by retesting of two additional subaliquots. Confirmed-positive specimens were tested for the presence of anti-B19 immunoglobulin M (IgM) and IgG with FDA-licensed assays. RESULTS: B19 DNA prevalence was 0.88 percent (95% CI, 0.64%-1.2%). Among the 23 donations with B19 DNA titers of at least 20 IU per mL, the median DNA concentration was 105 IU per mL with an interquartile range of 42 to 481 IU per mL; the highest value was 1869 IU per mL. All B19 DNA-positive donations were positive for the presence of IgG and 10 (23%) were also positive for the presence of IgM; IgM seropositivity was associated with increasing DNA levels (p = 0.0013). CONCLUSION: Low-level B19 DNA was detected in nearly 1 percent of donations. The 23 percent of DNA-positive donations with both IgM and IgG B19 antibody most likely represent acute resolving infection, whereas those with IgG but no IgM are most consistent with a more chronic and possibly persistent phase of B19 infection.     

Prevalence and Quantity of Parvovirus B19 DNA Among Blood Donors from a Regional Blood Center in Turkey

ObjectiveParvovirus B19 causes a range of diseases and morbidity in humans and is transmissible by transfusion of blood, blood components and plasma derivatives. The objective of the study was to investigate the prevalence and quantity of B19 DNA among blood donors. Method: Totally 1053 samples were collected from March to July 2016 at a blood bank for detection of Parvovirus B19 DNA and serological status of blood donors. Testing of the presence of viral DNA was performed by a quantitative real-time PCR with a 101 copies/ml detection limit. All DNA positive and randomly selected 267 samples were tested for the presence of anti-B19 IgM and IgG by ELISA.ResultsAge distribution of donors was between 18-64; mean age was 27 and median was 23. Among the 1053 samples, 5 (0.47%) had PB19 DNA. All PB19 DNA positive donations had both B19 IgM and IgG antibodies. The DNA level for positive donations were between 0.9 × 102 to 3.1 × 104 copies/ml. IgG and IgM were present in 59.9% (160/267) and 0,74% (2/267) respectively among the healthy donors without PB19 DNA.ConclusionDetected DNA concentration was less than 105 copies/ml. The presence of IgM in low level PB19 DNA positive donors may indicate that there might be a risk in transmission of PB19 to particularly immunosuppressed recipients. The clinical follow-up of blood donation with low level of PB19DNA should be considered to answer the questions about blood safety.     

Transfusion-transmitted human parvovirus B19 infection in a thalassemic patient

BACKGROUND: Human parvovirus (HPV) B19 infection has been shown to be transmissible by clotting factor concentrates, most often resulting in asymptomatic seroconversion. So far, no case of B19 transmission due to single-donor transfusion has been documented. CASE REPORT: A case of transfusion-transmitted HPV B19 infection in a 22-year-old female thalassemia major patient is described. She presented with an aplastic crisis; this was followed 1 week later by transitory heart failure and acute tricuspid incompetence. The echocardiogram revealed a grade III tricuspid regurgitation and a floating vegetation on the atrial face of the tricuspid lateral leaflet. The tricuspid regurgitation and vegetation spontaneously disappeared within 15 days. Blood cultures for bacteria were repeatedly negative. IgM anti-HPV B19 seroconversion was documented in the acute phase. B19 DNA was detected by polymerase chain reaction and remained detectable up to 4 months after diagnosis. High- titer IgM anti-HPV and B19 DNA were also found in serum samples collected at the time of donation from one of the donors of the blood transfused before the onset of clinical symptoms. CONCLUSION: This case documents the transmission of HPV B19 by the transfusion of 1 red cell unit and the occurrence of possible transient cardiac involvement in this infectious complication.