Quality of transfusion practice beyond the blood transfusion laboratory is essential to prevent ABO-incompatible death.
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Biomedical subjects
Publications and source records attributed to J F Chapman.
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The crossmatch has evolved from including a wide range of techniques through a test purely to eliminate ABO incompatibility (immediate spin) to computer crossmatching in which no serological testing is carried out and validation ensures the correct ABO/RhD type blood is issued. The crossmatch was always considered to be the most important feature of the compatibility test and in particular the antiglobulin phase; however, there are potential risks associated with serological and computer crossmatching including technical and procedural errors. The use of immediate spin and computer crossmatch change the emphasis for safety of the compatibility test from the crossmatch to the antibody screen. UK guidelines have now been published describing the features necessary for the introduction of computer crossmatching. Computer crossmatching is used by many institutions in various countries. It is considered safe practice and brings benefits to the laboratory and the patient. Compatibility testing is only one element of the blood transfusion procedure; the others are equally as important and include correct patient identification at the time of collection of the blood sample and at the administration of the blood transfusion.
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We have evaluated refractive index-matched anomalous defraction (RIMAD) (Dubin SB, Clin Chem 1988;34:938-43) as a potential method for assessment of fetal lung maturity (FLM). This method determines the total light scattered by the surfactant-containing lamellar bodies by subtraction of the A650 from amniotic fluid diluted in glycerol from that of amniotic fluid diluted in distilled water. It is not significantly affected by such contaminating chromogens as hemoglobin and bilirubin up to 2.0 g/L and 11.0 mg/L, respectively. However, the addition of as little as 2.5 microL of erythrocytes as whole blood resulted in significant interference. RIMADs for normal respiratory outcomes (n = 78) ranged from 0.018 to 0.471. RIMADs for respiratory distress syndrome (RDS) outcomes (n = 8) ranged from 0.004 to 0.036. Use of a RIMAD referent value of > 0.040 to indicate maturity yielded sensitivity, specificity, predictive value (PV)RDS, and PVmaturity of 100%, 96.2%, 72.2%, and 100%, respectively. The areas under the receiver-operating characteristic curves were 0.997 for the RIMAD assay, 0.993 (P = 0.3) for the TDx-FLM assay, 0.89 (P = 0.017) for the lecithin/sphingomyelin ratio, and 0.87 (P = 0.023) for the foam stability index.
OBJECTIVE: To assess the usefulness of the recently introduced TDx-FLM assay in managing pregnant women with diabetes. METHODS: Participating institutions were recruited from the 1993 and 1994 Society of Perinatal Obstetricians Diabetes Special Interest Group meetings. Study patients consisted of insulin-dependent diabetic women who had undergone transabdominal amniocentesis with assay of the fluid by the TDx-FLM method. Pertinent data were requested concerning pregnancy and respiratory outcomes of the corresponding neonates. RESULTS: Data from 261 pregnancies at 13 institutions were collected. Eight of the 182 infants born within 4 days of amniocentesis developed respiratory distress syndrome (RDS); five of the eight infants with RDS required intubation, and all five had TDx-FLM values less than 70 mg of surfactant per gram of albumin. Three of the eight infants with RDS required hood oxygen only; two of these infants had TDx-FLM values at least 70 mg/g. Thirteen of 144 (9%) subjects who delivered within 4 days of amniocentesis and for whom a TDx-FLM assay and phosphatidylglycerol level were both reported had a TDx-FLM level of at least 70 mg/g and a negative phosphatidylglycerol result. No infant with this combination of results developed RDS. Fifteen of the 40 patients who delivered more than 4 days after amniocentesis, with both tests available, had TDx-FLM values at least 70 mg/g and were phosphatidylglycerol negative. CONCLUSION: In infants of diabetic mothers, TDx-FLM values at least 70 mg/g were not associated with RDS requiring intubation. The TDx-FLM assay may be useful in determining the best time of delivery for pregnant patients with diabetes, especially in a situation in which the TDx-FLM assay is mature and the phosphatidylglycerol result is immature.
Amplification of specific gene target sequences has become a routine molecular procedure in a variety of laboratories. When coupled with either a direct or indirect method of detecting amplified product, in situ amplification offers an extremely powerful investigative tool. We describe a protocol for a localized in situ amplification (LISA) reaction that includes tissue-culture cloning rings and allows for the amplification of gene target sequences in specific regions of paraffin-embedded tissue sections. Digoxigenin-11-dUTP was added to the amplification reaction and thus incorporated into the amplified products, providing a mechanism by which direct nonisotopic detection could be performed. To demonstrate the approach, LISA was performed on known positive Pneumocystis carinii rat lung tissues, with primers specific for the P. carinii rRNA gene sequence.
OBJECTIVE: We evaluated the usefulness of the recently introduced TDx FLM assay in determining fetal lung maturity. STUDY DESIGN: The TDx FLM assay was compared with the lecithin/sphingomyelin ratio, the foam stability index, and the presence of phosphatidylglycerol in amniotic fluid samples from 102 pregnancies, among which respiratory distress syndrome developed in 22 infants. RESULTS: The TDx FLM assay met or exceeded results of other tests with respect to sensitivity (100% vs 86% to 100%), specificity (82% vs 38% to 78%), predictive value of a mature test result (100% vs 95% to 100%), predictive value of an immature test result (61% vs 31% to 51%), and efficiency (86% vs 51% to 79%). In a sequential testing strategy the TDx FLM assay performed well as the initial test. Our data indicate that the value for maturity suggested by the manufacturer may be more conservative than necessary. CONCLUSIONS: The TDx FLM assay appears to be a useful test in the field of fetal lung maturity testing.
Results reported here show that humans have various levels of trypanocidal activity in their sera. This difference appeared stable when different samples were taken from the same individuals over time. It was not possible to account for the variability between individuals by obvious differences in health, nutrition, or living habits. In addition, the trypanocidal titers did not vary significantly when stored for various lengths of time at -70 C. To examine the relationship between the titer of trypanocidal activity in a host and the degree of human serum resistance of the challenge trypanosome inoculum, mice (C57BL/6J) were pretreated with various amounts of different human serum and then infected with clones having different degrees of resistance to human serum. It was demonstrated that host susceptibility to an African trypanosome infection depends upon 2 variables: the level of trypanocidal activity in individual human serum and the degree of human serum resistance of individual clones of African trypanosomes. Based upon the animal model presented here, it is hypothesized that this relationship is under selective evolutionary pressure and will influence the susceptibility of animals in endemic areas as well as the transmission of human trypanosomiasis.
In 50 gravidas with insulin-dependent diabetes mellitus, mean preprandial whole blood glucose levels over a two-week period were compared with serum glycosylated hemoglobin (HbA1c) and fructosamine values. Although there was a statistically significant correlation between mean glucose levels and both HbA1c (r = .44, P less than .01) and fructosamine (r = .37, P less than .01), the wide range of HbA1c and fructosamine observed at all levels of mean blood glucose limited the usefulness of those assays in the management of pregnant diabetics.
Alloimmune neonatal thrombocytopenia (ANT) may cause intracranial haemorrhage in utero as well as at delivery. Recent management has concentrated on attempts to minimise fetal thrombocytopenia and prevent its complications. This report describes further experience with the use of repeated intravascular transfusions of compatible platelets in utero. The patient studied had already had one infant with intracranial haemorrhage due to ANT. In her next pregnancy, weekly intra-uterine platelet transfusions were given from 26 weeks, but intra-uterine death occurred at 30 weeks after the mother had a heavy fall. In her most recent pregnancy, weekly intravascular transfusions of platelets were given by cordocentesis from 29 to 34 weeks. The fetal platelet count was maintained above 30 X 10(9)/l for almost all of the last 6 weeks of pregnancy before delivery of a normal infant by Caesarean section at 35 weeks' gestation. This approach is effective in preventing severe fetal thrombocytopenia in the last trimester of pregnancy and is contrasted with alternative treatments of ANT. Further data are required to determine the efficacy and risks of these treatments.
We evaluated the performance of the PG-Numeric (PGN; Isolab, Inc.) enzymatic assay for phosphatidylglycerol (PG) adapted for semi-automated analysis on the Cobas-Bio (Roche) centrifugal analyzer. We evaluated precision, linearity, and potential interference from endogenous glycerol. In addition, we compared the results of the PGN assay with those for two other PG assays. Amniostat-FLM (AFLM; Hana Biologics) and phosphatidylglycerol/sphingomyelin ratio (PG/S), on amniotic fluid samples from 51 patients. Within-run and total CVs for the PGN test (n = 80) were respectively 8.6% and 11.4% for PG at 2.6 mumol/L, and 4.5% and 6.2% at 7.3 mumol/L. The PGN test exhibited no significant deviation from linearity between PG concentrations of 0.4 and 15 mumol/L. Endogenous glycerol up to 500 mumol/L did not affect PGN results significantly. Results of the PGN assay compared favorably with those of the other PG assays for tests of fetal lung maturation. The Cobas-Bio adaptation of the PGN test performed well analytically and clinically, and allowed the determination of PG in less than 1 h.
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The occurrence of post-transfusion purpura (PTP) in a 16-year-old girl with sickle/beta-thalassaemia is described. Clinically this was a typical case of PTP, but it was unusual serologically. Anti-Baka and anti-PIA2 platelet-specific antibodies were identified and the patient's platelets were typed as homozygous PIA1-positive and Baka-negative. The patient also developed red-cell, granulocyte and lymphocytotoxic antibodies in response to the blood transfusion and had a delayed haemolytic transfusion reaction.
Performing multiple tests of fetal lung maturity on amniotic fluid samples may not use the individual test results and laboratory personnel most effectively. To determine the best strategy for fetal lung maturity testing, we analyzed our experience with use of a variety of procedures. Clinical usefulness was assessed according to sensitivity, specificity, predictive values, and efficiency. Economic and technical aspects analyzed included time and personnel requirements, availability of tests, and expense of each procedure. Several testing sequence approaches were compared for efficiency and cost. In our laboratory the foam stability index proved to be the most useful initial test of fetal lung maturity, reserving more expensive and time-consuming tests for instances in which the foam stability index is immature. Routine multiple testing did not enhance clinical usefulness and greatly increased costs. Development of a testing strategy using a rapid, inexpensive, and widely available test such as the foam stability index would promote clinical and economic efficiency.
Patients with acute leukaemia who are exposed to intensive chemotherapy, with or without radiotherapy, may remain thrombocytopenic or neutropenic in remission. The incidence of these prolonged cytopenias was examined retrospectively in 46 patients in remission and prospectively in 14 patients. The patients were tested for the presence of autoantibodies to platelets and neutrophils using a fluorescent antihuman globulin technique with paraformaldehyde-fixed cells. In the retrospective study nine patients (20%) had neutrophil autoantibodies and seven (15%) had platelet autoantibodies; only one of the former had neutropenia and one of the latter thrombocytopenia. In the prospective study three (21%) had neutrophil autoantibodies and seven (50%) had platelet autoantibodies. One of the patients with platelet autoantibodies had transient thrombocytopenia, and shortened platelet survival was demonstrated. None of the patients with neutrophil autoantibodies had neutropenia. The rare occurrence of cytopenias in association with these autoantibodies was possibly due to bone marrow compensation for antibody-mediated cell destruction, although other possible mechanisms are discussed.
Uniform liquid-chromatographic conditions were developed such that we could quantify norepinephrine, epinephrine, normetanephrine, metanephrine, vanillylmandelic acid, and 5-hydroxyindoleacetic acid in urine by using a single mobile phase of monochloroacetic acid and citric acid, 0.1 mol/L each. All compounds were separated on a C18 column and detected electrochemically at a potential of +0.800 V. Optimization of these uniform chromatographic conditions significantly shortens the changeover time required from one assay to another, resulting in a substantial savings of time and cost to the laboratory.
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This study describes the development of autoimmune thrombocytopenia followed by autoimmune haemolytic anaemia in a Negro woman with measles. An IgM platelet autoantibody was detected using a fluorescent labelled antiglobulin technique. The thrombocytopenia resolved spontaneously, although the platelet autoantibody persisted and platelet survival remained shortened, suggesting a compensated thrombocytolytic state. An IgG granulocyte autoantibody was present transiently although the patient was never neutropenic. The haemolytic anaemia was due to an IgM cold autoantibody (anti-I), which was active up to 30 degrees C, and an IgG warm autoantibody, which was detectable only when she was severely anaemic. After an initial blood transfusion, the anaemia resolved and the red cell autoantibodies disappeared. The platelet, granulocyte and red cell autoantibodies were cell-specific and not a single cross-reacting antibody.