First report of human immunodeficiency virus transmission via an RNA-screened blood donation.

BACKGROUND AND OBJECTIVES: Blood banks in the USA have recently introduced minipool nucleic acid amplification testing (MP-NAT) of blood products to reduce the transmission of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) by transfusions. However, MP-NAT is limited in its ability to detect preseroconversion samples with very low viral RNA loads. MATERIALS AND METHODS: To determine whether a red blood cell unit, from an MP-NAT-negative donation, transmitted HIV when transfused to a patient, we compared the viral sequences from the blood donor and recipient. The implicated donation was also tested by commercially available NAT assays at a range of dilution factors to determine whether the infectious unit could have been detected using individual-donation NAT (ID-NAT). RESULTS: Phylogenetic linkage of HIV sequences in the blood donor and recipient confirmed the transmission of HIV by blood transfusion, the first such case identified since introduction of MP-NAT screening in 1999. Viral RNA was reliably detected by ID-NAT, but only inconsistently detected by MP-NAT. CONCLUSIONS: Even following the introduction of MP-NAT, a preseroconversion donation with a viral load of

ISO 9000 model ideally suited for quality plan at blood centers.

BACKGROUND: The development of a quality plan is essential for organizations involved in manufacturing, trade, and services. The International Organization for Standardization (ISO) has established a common set of manufacturing, trade, and communications standards that are applicable worldwide and that provide the basis of a quality plan for institutions such as blood centers. STUDY DESIGN AND METHODS: The ISO 9002 conformance model and a registrar were selected to guide a core management team in establishing a superior quality plan for a blood center. The initial phase required that an analysis of the existing quality and document systems be performed, utilizing a 20-element "status of readiness" ISO assessment audit (gap analysis). Weaknesses in the quality systems were targeted and progress was evaluated by the registrar and an outside facilitator. RESULTS: Full implementation of ISO principles was achieved within the established timeline. Standard operating procedures were reviewed, supplemented, and expanded in every department, thereby ensuring consistent quality throughout the organization. Improvements in management, training, inspections, and statistical reporting were soon apparent during regular departmental audits. CONCLUSION: The ISO 9000 model is ideally suited for use by blood centers to establish a quality plan as required by the American Association of Blood Banks.

An effective method for the preparation of leukocyte-poor platelets.

Febrile transfusion reactions are encountered occasionally in patients receiving platelet concentrates that contain contaminating leukocytes. To remove heavier cellular elements, a 400-ml platelet pooling bg with tapered sides and a pouch at the base was designed to trap these cells after centrifugation. Two versions of this bag were tested: the initial prototype with a 3-ml pouch and a modified version with a 4-ml pouch. Platelet concentrates were pooled in batches of 6 per bag and, after sampling, were centrifuged between 186 and 600 x g for 7 to 10 minutes. The pouch on each bag was then clamped off, and samples from the primary bag were withdrawn so tht the percentage of cellular elements remaining could be determined. At optimal centrifugation conditions (390 x g for 10 minutes) with the initial prototype of the bag, average cellular decreases were: leukocytes, 75.9 percent; red cells, 84.1 percent; and platelets, 9.8 percent. Twenty-two components prepared in this manner were infused into 12 patients with histories of febrile transfusion reactions. Febrile reactions were markedly reduced or absent in ten patients, a strong febrile reaction due to incorrect component preparation occurred in one, and a marked allergic reaction occurred in one. The 4-ml pouch gave much better production results with greater than 95 percent leukocyte removal and less than 5 percent platelet loss. This system removes sufficient leukocytes from platelet concentrates that the risk of febrile transfusion reactions is reduced significantly.

A microagglutination test for blood typing rhesus monkeys, Macaca mulatta.

We have developed a microagglutination test for typing rhesus monkey erythrocytes that is sensitive, accurate and easy to perform. The technique requires only microliter quantities of antiserum and cells, and agglutination is easily detected using an inverted microscope. An advantage of this technique is that the typing plates can be stored at -70 degrees C without loss of activity. The results of typing over 400 rhesus blood samples with this technique were 95% concordant with results using the standard microtitre agglutination technique. Preliminary results indicate that this test is also adaptable to typing human blood.

Preparation of granulocyte concentrates for neonatal patient transfusion.

As an adjunct to antibiotic therapy, granulocytes for infusion often are required within hours of the diagnosis of neonatal sepsis. The logistics of obtaining granulocytes by leukapheresis were evaluated in order to provide, at short notice, a low volume, highly concentrated product. An average of 4.0 X 10(9) granulocytes in a mean volume of 101.5 ml of plasma was collected after processing an average of 3 liters of blood from each of three donors. Following centrifugation of these products, the volume of the final products was reduced by an average of 54.3 percent, while only 10.2 percent of the granulocytes were lost. One of the products, containing 4.8 X 10(9) granulocytes in a volume of 44 ml, was infused into a 3.4 kg septic neonatal patient within 4 hours of receipt of the order by the blood bank. We conclude that it is possible to collect high concentrations of granulocytes for neonatal transfusion in a timely manner by following the protocol described in this report.

In vitro assessment of platelet damage during rotator storage.

The measurement of beta-thromboglobulin (BTG) and platelet lactic dehydrogenase (LDH) as markers for alpha-granule release and platelet lysis, respectively, has been reported to correlate with the amount of damage incurred during the preparation and storage of platelet concentrates. We compared the concentrations of these markers in the supernatant plasma of platelets stored on platform, elliptical, and circular rotators. Significant BTG release occurred after 24 hours on all three rotators (P less than 0.01), but LDH discharge after 24 hours was significant (P less than 0.05) only with elliptical rotation. BTG and LDH levels were not significantly different at any time when platform and circular rotations were compared, but both markers were significantly (P less than .05) higher during elliptical rotation. The lower levels of BTG release and LDH discharge during platform and circular agitation implies that these rotators produce less in vitro platelet activation and damage than occurs with elliptical agitation.

Comparison of two continuous-flow cell separators.

Two blood processors (IBM 2997 and Fenwal CS-3000) were evaluated under similar conditions. Fifty-four leukapheresis procedures with the 2997 resulted in a mean granulocyte yield of 19.4 X 10(9) (42.5% efficiency), with a mean of 2.1 X 10(11) platelets (10.9% efficiency) per product. The CS 3000, at a whole blood flow rate of 50 ml/min, yielded a mean of 13.3 X 10(9) granulocytes (39.2% efficiency) and 4.0 X 10(11) platelets (28.5% efficiency) during 63 leukapheresis procedures. At a flow rate of 60 ml/min, the mean yields of 20 leukapheresis procedures with the CS 3000 were 14.2 X 10(9) granulocytes (30.5% efficiency) and 4.3 X 10(11) platelets (27.8% efficiency). Thirty-four plateletpheresis procedures with the 2997 yielded a mean of 3.62 X 10(11) platelets (53.12% efficiency), and 2.70 X 10(9) white cells. The mean CS-3000 yield for 88 plateletpheresis procedures was 3.15 X 10(11) platelets (49.13% efficiency) with a mean white cell content of 0.67 X 10(9). Granulocyte yields with the 2997 were greater than those obtained with the CS-3000.

Plasmapheresis in a child with the hemolytic-uremic syndrome.

Plasmapheresis, using fresh-frozen plasma for replacement, has been reported to be of benefit in the treatment of hemolytic-uremic syndrome and in thrombotic thrombocytopenic purpura. Both diseases have the same pathological characteristics and, possibly, the same pathogenesis. The mechanism by which plasma exchange results in clinical improvement is not clearly established, although recent reports indicate that it may be based on prostacyclin replacement rather than the removal of humoral factors toxic for platelets or vascular walls. Exchange plasmapheresis using an intermittent-flow cell processor with a disposable 100-ml pediatric centrifuge bowl is described in a 5-year-old girl with hemolytic-uremic syndrome.

Platelet washing with a blood cell processor.

Washing of platelet components for transfusion may be indicated in selected patients. This study was designed to determine a protocol using an automated cell processor to remove plasma proteins from pooled platelet concentrates, stored at 20 to 24 degrees C with constant rotation. A 1500-ml, 0.9% saline-wash procedure is described. Eight percent of the platelets were lost with the procedure (range, 0-19%), while a mean of 99.6 percent of the plasma protein was removed (range, 99.2-99.8%). The clinical effectiveness of maternal platelets washed by this method was demonstrated in a case of alloimmune neonatal thrombocytopenia.

Relative values of laboratory assays in systemic lupus erythematosus.

The sensitivities and the specificities of Crithidia luciliae immunofluorescence, (CL-IF), counterimmunoelectrophoresis (CIE), and enzyme-linked immunosorbent assay (ELISA) as aids to the clinical diagnosis of systemic lupus erythematosus (SLE) were compared to determine which was the single most useful test in the management of this disease. The patients who had SLE were further divided into those with active or inactive disease and those with and without nephritis in an attempt to determine whether any of these tests could reliably differentiate these groups. Results obtained by these tests, together with those of a nonspecific antinuclear antibody assay used as a screening test, were compared with results for other autoimmune diseases. Although the ELISA was more often positive than CIE and CL-IF in cases of SLE, CL-IF had the highest specificity for SLE, giving no positives in any of the other autoimmune diseases examined. None of the tests satisfactorily differentiated active from inactive SLE or consistently detected the presence or absence of nephritis in SLE.