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Biomedical subjects

K Sazama

Publications and source records attributed to K Sazama.

At least 37 records · Page 2Linked to original sources

Hepatitis C virus in blood samples from volunteer donors.

The specificities of four assays for hepatitis C virus (HCV) were compared by using units from volunteer blood donors. Upon Food and Drug Administration licensure of the first immunoassay for anti-HCV, EIA-1, units previously deemed acceptable for transfusion and all subsequent blood donations were screened. EIA-1 repeat-reactive (RR) units were tested for HCV by a second-generation enzyme-linked immunoassay (EIA-2) and by a four-antigen recombinant immunoblot assay (RIBA II) and for HCV RNA by reverse transcriptase polymerase chain reaction. All HCV RNA-positive samples were reactive by both RIBA II and EIA-2. All RIBA II-reactive sera were EIA-2 RR. In EIA-1, 0.45% of the prescreened units and 0.59% of the prospectively screened donors were RR. Of these, 52.5 and 54%, respectively, were EIA-2 RR, 71.4 and 69% of the EIA-2 RR units were reactive on RIBA II, and 93 and 88% of the RIBA II-reactive samples were HCV RNA positive. When the sample/cutoff ratio for EIA-2 was greater than 5, 91% of the samples were RIBA II reactive and 82% of the samples were HCV RNA positive. None of EIA-2 RR units with a sample/cutoff ratio of < 5 was RIBA II reactive or HCV RNA positive. In conclusion, RIBA II and RT PCR results are highly concordant. A sample/cutoff ratio of > 5 in EIA-2 is a good discriminator for the likelihood of a true HCV infection on the basis of RT PCR and RIBA II assays.

Base Sequence↗

Detection of antibodies to human immunodeficiency virus type 2 (HIV-2) in blood donor sera using United States assay methods for anti-HIV type 1.

Twelve serum samples from French blood donors that were uniformly reactive in tests for antibody to human immunodeficiency virus type 2 (anti-HIV-2) also were reactive in 92 to 100 percent of tests with three anti-HIV type 1 (anti-HIV-1) enzyme-linked immunoassays currently in widespread use for donor screening in the United States. Supplemental tests for anti-HIV-1 on these anti-HIV-2-reactive samples differed in their responses. All samples reacted in a licensed anti-HIV-1 Western blot, but there was an atypical band near the p41 position, which could be a clue to the fact that this result was a cross-reaction with anti-HIV-2. A recombinant immunoblot gave an indeterminate result for anti-HIV-1 in all 12 samples. A local immunofluorescence assay for anti-HIV-1 reacted with 92 percent of the samples, but a commercial one detected only 58 percent.

Antibodies, Viral↗

Detection of antibody to immunodeficiency viruses by dot immunobinding assay.

The dot immunobinding assay (DIA), a modified enzyme immunoassay (EIA), has been demonstrated to be a highly sensitive and specific assay for the detection of antibody to a number of viruses. Different laboratory procedures are available for detecting antibody to the immunodeficiency viruses; however, these procedures require a certain amount of sophisticated equipment and trained personnel. Further, commercial kits for detecting antibody to human immunodeficiency virus, as now available, are not easy to use in the nonlaboratory setting. The DIA, as described herein, may be formatted to test up to 30 serum samples and is designed to be used in the absence of laboratory equipment. To determine the effectiveness of the DIA as a test kit for the detection of HIV and human T-cell leukemia virus type I (HTLV-I) antibodies, the kit was compared with commercial EIA and Western blot (WB; immunoblot) kits. Testing approximately 1,000 human serum samples for HIV antibody by DIA and EIA revealed a total agreement of 98.1%, a specificity of 99.0%, and a sensitivity of 95.9%. For 804 serum samples tested (200 were tested independently in two laboratories), eight results were discrepant: four DIA negatives which were EIA borderline positive and four DIA positives which were EIA negative. Testing the eight discrepant sera by immunofluorescence assay and WB resulted in their being either negative or indeterminate. The four DIA positives were indeterminate by WB. Close agreement was obtained when the remaining sera were compared by DIA, EIA, and WB. Of interest was finding that the DIA results compared favorably with those obtained by WB. Twenty-six suspect HTLV-I-positive serum samples tested by DIA also gave results comparable to those obtained by EIA and WB.

Blotting, Western↗

Seroepidemiology of viral infections among intravenous drug users in northern California.

Intravenous drug users are frequently exposed to parenterally transmitted viral infections, and these infections can spread to the general population through sexual activity. We investigated the prevalence of serologic markers for human immunodeficiency virus type 1 (HIV-1), human T-cell lymphotropic virus type I/II (HTLV-I/II), hepatitis B virus (HBV), and hepatitis C virus (HCV) in intravenous drug users and their sexual contacts. Of 585 drug users from northern California tested for these serologic markers, 72% were reactive for the antibody to HCV, 71% for the antibody to hepatitis B core antigen, 12% for HTLV-I/II antibodies, and 1% for the HIV-1 antibody. The prevalence of serologic markers for these four viruses correlated with the duration of intravenous drug use, the ethnic group, and the drug of choice. More than 85% of subjects infected with either HCV or HBV were coinfected with the other virus. All persons reactive to HTLV-I/II antibodies had antibodies for either HBV or HCV. Of 81 sexual contacts tested, 17% had evidence of HBV infection while only 6% were reactive for HTLV-I/II antibodies and 4% for the antibody to HCV. None of this group was infected with HIV-1. We conclude that HTLV-I/II and HCV are inefficiently transmitted to sexual contacts while HBV is spread more readily. Programs designed to discourage the sharing of drug paraphernalia, such as needle and syringe exchanges, should decrease the risk of parenterally spread viral infections in intravenous drug users and thus slow the spread of these infections to the general population.

Adult↗

Reports of 355 transfusion-associated deaths: 1976 through 1985.

From 1976 through 1985, the United States Food and Drug Administration received reports of 355 fatalities associated with transfusion, 99 of which were excluded from further review because they were unrelated to transfusion or involved hepatitis or acquired immune deficiency syndrome. Of the remaining 256 reported deaths, 51 percent resulted from acute hemolysis following the transfusion of ABO-incompatible products. These deaths were due primarily to managerial, not clerical, errors. Other causes of death (in order of frequency of report) included acute pulmonary injury (15%), bacterial contamination of product (10%), delayed hemolysis (10%), damaged product (3%), and graft-versus-host disease (0.4%). Management systems for transfusion facilities should be created or revised to include the specific identification of personnel eligible to administer transfusions to provide written guidance and appropriate training (including recognition and management of errors), and to implement measures that target safe transfusion practices. Continued research into acute pulmonary injury, the immunologic hazards of transfusion, and the prevention of bacterial contamination of blood components is necessary.

Acute Disease↗

Stat testing and DRGs: a strategy for cost containment.

To reduce costs in the laboratory we must determine when stat testing is appropriate and establish a monitoring system. Here, as a means of reviewing the use of such tests and as an aid in decision making, is a break down of the probability of stat testing based on Diagnosis-related Group categories.

Clinical Laboratory Techniques↗

Intraoperative hemolysis. The initial manifestation of glucose-6-phosphate dehydrogenase deficiency.

Unexpected intraoperative hemolysis suggestive of a hemolytic transfusion reaction was the initial clinical manifestation of Mediterranean type glucose-6-phosphate dehydrogenase (G-6-PD) deficiency in a 50-year-old Indian man. There was no evidence for immune mediated hemolysis since no unexpected RBC antibodies were found, and results of the direct antiglobulin test remained negative. Analysis of preoperative blood specimens demonstrated complete absence of G-6-PD activity. No etiologic agent clearly associated with G-6-PD hemolysis could be identified. Testing for G-6-PD deficiency should be considered when unexpected hemolysis occurs intraoperatively.

Glucosephosphate Dehydrogenase Deficiency↗

Is antiglycolysis required for routine glucose analysis?

We obtained -68 pairs of simultaneously drawn serum and fluoride-oxalate plasma samples from patients and analyzed them by a continuous-flow (AutoAnalyzer II) glucose oxidase method. Glucose concentrations ranged from 370 to 3530 mg/L. Glucose concentrations for samples obtained in the fluoride-oxalate preservative averaged 42 +/- 35 mg/L (mean +/- SD) higher than serum. The magnitude of this difference was independent of glucose concentration. Linear-regression analysis of 270 pairs for which the time from collection to separation was recorded indicated that the difference between serum and plasma increased by 0.32 mg/L per minute of delay over a time span of 15 to 295 min. These differences are smaller than those described in standard textbooks. We conclude that, with the specimen-handling process used in our hospital, serum glucose determinations are clinically acceptable.

Blood Chemical Analysis↗

Detection of plasmapheresis-induced platelet activation using monoclonal antibodies.

Twelve volunteer plasma donors were studied so as to determine the extent and duration of in vivo platelet activation caused by automated plasmapheresis. Samples obtained immediately before and after donation were mixed with murine monoclonal antibodies PAC-1 and S12, which bind specifically to activated platelets. Antibody binding on platelets was quantitated by flow cytometry. The change in the mean fluorescence intensity (MFI) (MFI after donation minus MFI before donation) was 61 +/- 23 (confidence interval [CI], 48-75) for PAC-1 and 56 +/- 64 (CI, 19-92) for S12 in plasmapheresis donors, as compared to 0.3 +/- 0.8 (PAC-1; CI, -0.2-0.8) and 0.3 +/- 0.9 (S12: CI, -0.3-0.9) in whole blood donors (p less than 0.05). Additional studies showed circulating activated platelets up to 48 hours after plasmapheresis. In contrast to other data, significant platelet activation was demonstrated following plasmapheresis on an automated machine. None of the donors had clinical complications. Nevertheless, it may be appropriate to delay subsequent plasmapheresis and platelet procurement from such donors until evidence of platelet activation has disappeared.

Antibodies, Monoclonal↗