ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36596282579.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36596282580.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND:A recent hepatitis C virus (HCV) outbreak has been suspected of being caused by an infusion of intravenous immune globulin.STUDY DESIGN AND METHODS:Three laboratories were mandated by the French regulatory agency to prospectively screen on a national scale those persons having received suspected batches: 233 exposed patients were recalled and tested for HCV antibody and for HCV RNA.RESULTS:Nineteen patients (8.1%) were found positive for HCV RNA; 7 of these 19 were positive for the HCV antibody.CONCLUSION:The link between HCV infection and intravenous immune globulin was reinforced by the overrepresentation of the 2b genotype (58%), which contrasts with the low prevalence of this genotype in France (1%).

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36596282581.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND:Reports that the human immunodeficiency virus type 1 (HIV‐ 1) group O variants are not reliably detected by some commercial diagnostic tests have raised concerns about the sensitivity of existing screening tests, especially with regard to blood safety. Although it is unlikely that these divergent strains are prevalent in North America, systematic, continuous surveillance is needed to monitor the potential spread of HIV variants into that region.STUDY DESIGN AND METHODS:Stored serum samples (n = 1072) from both high‐ and low‐risk population groups at several sites in the United States and Puerto Rico were tested by peptide enzyme immunoassays specific for the prototypic HIV‐1 group O strains, MVP5180 and ANT70. RESULTS: None of the 1072 samples examined had peptide reactivity that was consistent with HIV‐1 group O infection.CONCLUSION:While no evidence of specific HIV‐1 group O (MVP5180 or ANT70) infection was found in this study, the sensitivity of current tests has not been fully evaluated against the wide range of genetic variation of HIV. Therefore, it is important to continue active surveillance for HIV‐1 and HIV type 2 strains, to characterize any divergent strains, and to judiciously modify tests to correct for any deficiencies i

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36596282582.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND:Reactive samples in hepatitis C virus (HCV) antibody screening of blood donors are currently referred for a confirmatory assay. This scheme is not optimally efficient and is expensive because of the lack of specificity and cost of confirmatory tests, as well as the need to discard false‐positive donations. As in some human immunodeficiency virus antibody‐confirmatory schemes, the safety and efficacy of confirming anti‐HCV with two sequential screening assays were evaluated.STUDY DESIGN AND METHODS:Three combinations of two anti‐HCV screening assays were used to test 75,874 blood donors. Results were compared with the routine testing scheme and HCV RNA detection in any enzyme immunoassay‐repeatably reactive samples.RESULTS:The use of an alternative screening assay for repeat testing decreased the proportion of enzyme immunoassay‐positive donors from 0.28 to 0.05 percent. All samples that were “confirmed” as positive by the standard combination of immunoassays and all HCV RNA‐positive samples were detected by the sequential screening assays. No samples that had discordant results on primary and secondary screening assays were confirmed by recombinant immunoblot assay or were found to contain detectable HCV RNA.CONCLUSION:The combination of screening assays for anti‐HCV confirmation was as safe as, cheaper than, and nearly as efficient as the stan

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36596338024.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND:Photochemical decontamination of platelet concentrates (PCs) has been demonstrated by the use of 8‐methoxypsoralen and ultraviolet A light. Systems for studying the inactivation of blood‐ borne viruses facilitate the evaluation of photochemical decontamination protocols.STUDY DESIGN AND METHODS:Duck hepatitis B virus (HBV), a model for human HBV, was adapted for the study of hepadnavirus inactivation. A highly specific in vitro infectivity assay used primary duck hepatocyte cultures and was followed by the detection of replicated duck HBV sequences. RESULTS: Duck HBV‐infected primary duck hepatocyte cultures produced authentic infectious virus. High‐ titer (>10(9) virus genome equivalents/mL) duck HBV‐infected sera were completely inactivated in serum or PCs by the use of 100 micrograms per mL of 8‐methoxypsoralen and 70 J per cm2 of ultraviolet A light. Intracellular duck HBV (>4.2 log10) in PCs was also inactivated. Culture results were confirmed by a sensitive duckling infectivity assay that indicated that 6.3 log10 of infectious duck HBV had been inactivated by photochemical decontamination.CONCLUSION:The sensitivity of the culture assay was comparable to that of the duckling assay using polymerase chain reaction gene amplification to detect duck HBV. Duck HBV inactivation in PCs was dependent on the dose of ultraviolet A light and independent of 8‐methoxypsoralen concentrations of 100 to 300 micrograms per mL: 100 micrograms per mL 8‐ methoxypsoralen inactivated 4 to 5 log10 of virus in conjunction with 20 to 40 J per cm2 of ultraviolet A light. The polymerase chain reaction‐enhanced duck HBV culture system has utility in optimizing photochemical decontam

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36596282583.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36596282584.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND. Previous studies established an association between the red cell chemokine receptor and Fy6 of the Duffy glycoprotein. The relationship between Duffy system antigens and interleukin 8 binding by red cells was examined.STUDY DESIGN AND METHODS. Interleukin 8 adsorption was measured by using red cells from owl, squirrel, and rhesus monkeys, and human red cells before and after saturation with anti‐Fya (anti‐Fy1), anti‐Fyb (anti‐Fy2), or anti‐Fy6. The effect of cell saturation with interleukin 8 on Duffy system antigen expression and the effect of enzyme treatment on interleukin 8 adsorption were also examined. RESULTS. Fy:1,‐2,6 or Fy:‐1,2,6 (owl monkey, squirrel monkey, human) red cells adsorbed interleukin 8 over a wide range of concentrations (125–1000 pg/mL). Human Fy:‐1,‐2,‐6 red cells bound minimal interleukin 8. Human red cells saturated with corresponding antibodies displayed decreased interleukin 8 binding. Saturation of human cells with interleukin 8 failed to interfere with Duffy system antigen expression. Rhesus monkey cells lacking Fy1, Fy2, and Fy6 were able to adsorb interleukin 8 in a manner similar to Fy:1,‐2,6 or Fy:‐ 1,2,6 primate cells. Ficin treatment eliminated both Duffy system antigen expression and interleukin 8 adsorption from red cells.CONCLUSIONS. The Duffy glycoprotein binds interleukin 8. Although previous studies suggested that interleukin 8 bound either directly to or close to the Fy6 epitope, the ability of Fy:‐1,‐2,‐6 rhesus monkey cells to adsorb interleukin 8 suggests that the interleukin 8 receptor is

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36596282585.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND:Platelet‐specific antibodies may be involved in refractoriness to platelet transfusions, disorders such as neonatal alloimmune thrombocytopenia, and post‐transfusion purpura. Genotyping for the major human platelet antigen (HPA) systems HPA‐1 through HPA‐5 is of considerable help in establishing the diagnoses of these diseases.STUDY DESIGN AND METHODS:A new genotyping method is described. Alleles of all five systems are amplified in a multiplex polymerase chain reaction. Subsequently, aliquots of the amplification products are thermocycled in the presence of a pair of allele‐specific oligonucleotide probes and a heat‐stable ligase. After heat denaturation, the probes hybridize adjacent to complementary sequences of the amplification product. In a perfect match, the two probes become covalently joined. Detection of the ligation product is performed with an enzyme‐linked immunosorbent assay.RESULTS:Complete concordance of genotypes between the ligation‐based typing and established genotyping methods was determined in 54 Austrian (HPA‐1, −2, −3, and −5) and 56 Japanese (HPA‐4) individuals. Ligation‐based genotyping of HPA‐1 polymorphism using platelet‐derived RNA as starting material gave concordant results in all 15 cases tested.CONCLUSION:Multiplex polymerase chain reaction in combination with ligation‐based typing allows fast typing of large numbers of platelet donors and screening for cr

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36596282586.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND:Determination of the volume of fetal D‐positive cells in the circulation of D‐negative women after delivery is carried out to determine whether additional prophylactic anti‐D should be given to the mother. Although the Kleihauer‐Betke test is still widely used to calculate the fetomaternal hemorrhage, increasing use is being made of flow cytometry.STUDY DESIGN AND METHODS:A conjugated monoclonal anti‐ D was prepared by labeling purified BRAD‐3 (IgG3) with fluorescein isothiocyanate (FITC‐BRAD‐3). This reagent was used to label D‐positive red cells by a one‐step procedure: 5 microL of washed cells were incubated with 50 microL of FITC‐BRAD‐3 (50 micrograms/mL) at 37 degrees C for 30 minutes; then the cells were washed and 500,000 events were analyzed by flow cytometry.RESULTS:The FITC‐BRAD‐3 reagent effectively labeled D‐positive cells. The percentage of D‐positive cells in mixtures containing more than 0.04 percent D‐positive cells in D‐negative cells was accurately determined by using this reagent and flow cytometry. Although the Kleihauer‐Betke test was more accurate than this one‐step flow cytometric method at quantifying fetomaternal hemorrhage of<1 mL, the flow cytometric method was more accurate in the 1‐ to 7‐mL fetomaternal hemorrhage range of 1 to 7 mL (whole‐blood equivalents). Analysis of 175 clinical samples for fetomaternal hemorrhage gave consistent quantification results with the three methods used: the Kleihauer‐Betke test, flow cytometry with FITC‐BRAD‐ 3, and flow cytometry with polyclonal anti‐D followed by FITC‐anti‐IgG.CONCLUSION:Labeling of samples with FITC‐BRAD‐3 was simple and rapid. By flow cytometric analysis, good separation of D‐positive from D‐ negative cells was obtain

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36596282587.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY