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Which techniques should be used to screen blood donations for hepatitis B surface antigen?

Preservation of all donor sera already tested for Hepatitis B surface antigen (HBsAg) by counterimmunoelectrophoresis (C.I.E.P.) allowed C.I.E.P., reversed passive haemagglutination (R.P.H.A.), and radioimmunoassay (R.I.A.) to be evaluated as screening techniques. Out of 165 811 donors tested for the first time 207 were found by C.I.E.P. to be positive for HBsAf-a prevalence of 0-12%. At the next donation 10 of those apparently negative on first screening were HBsAg positive by C.I.E.P., and nine of these were shown by retesting with R.P.H.A. and R.I.A. to have been positive at the earlier donation. These nine false negatives caused four cases of transfusion-transmitted HBsAg-positive hepatitis. On comparative screening of 22 239 donations C.I.E.P. detected 27 sera positive for HBsAg, R.P.H.A. 39, and R.I.A. 41. Thus R.I.A. increased the detection rate by more than half over C.I.E.P. From these 14 further false-negative donations by C.I.E.P. six cases-one fatal-of HBsAg-positive hepatitis occurred. The number of false positive reactions when using R.P.H.A. or R.I.A. for screening was less than 1%. As many as 700 donations in one day have been tested by R.P.H.A. or R.I.A. R.P.H.A. is faster and less expensive than R.I.A., but R.I.A. is more objective. Either R.P.H.A. or R.I.A. should replace C.I.E.P. as the routine method of screening donor sera for HBsAg.

Blood Donors↗

Limulus amebocyte lysate testing of nomral serum albumin (human) in the United States since 1975.

In order to determine the reactivity of Limulus Amebocyte Lysate to albumin products, over 1500 lots of 25% Normal Serum (Human) produced by 12 U.S. licensed manufacturers were tested over the past 4 years. Representative samples of 5% Normal Serum Albumin (Human) and Purified Protein Fraction (human) were also tested with Limulus Amebocyte Lysate. Problems associated with apparent false negative reactions are discussed. Model test protocols are presented that might be useful in determining the degree of false positivity of albumin products. These models could be adopted by blood fractionators to gather validation data to support substitution of the Limulus Amebocyte Lysate test for the rabbit pyrogen test.

Animals↗

Complement fixation antibody test for human nocardiosis.

Complement-fixing antibody was detected in 13 of 16 patients with histological and/or culture evidence of infection with Nocardia species. The antigen used was a filtrate of soluble substances secreted into liquid growth medium by cultures of N. asteroides. Apparent false positives reactions were found in three of three patients with leprosy and two of five patients with tuberculosis--results similar to some previously reported methods. N. asteroides and mycobacteria share antigens. Only the false positives with tuberculosis are considered a diagnostic problem. No reactions were obtained with sera from 26 patients with other infections and 41 unifected individuals. Whereas previous nocardia serodiagnostic methods have a sensitivity of approximately 50%, our overall sensitivity (81%) compares favorably and included 9 of 11 positive tests in immunocompromised nocardiosis patients (a source of false negative reactions with previous methods).

Adult↗

Discrepancy between culture and DNA probe analysis for the detection of periodontal bacteria.

The purpose of this study was to compare a commercially available DNA probe technique with conventional cultural techniques for the detection of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in subgingival plaque samples. Samples from 20 patients with moderate to severe periodontitis were evaluated at baseline and during a 15 months period of periodontal treatment. Paperpoints from 4 periodontal pockets per patient were forwarded to Omnigene for DNA probe analysis, and simultaneously inserted paperpoints from the same pockets were analyzed by standard culture techniques. In addition, mixed bacterial samples were constructed harbouring known proportions of 25 strains of A. actinomycetemcomitans, P. gingivalis and P. intermedia each. A relatively low concordance was found between both methods. At baseline a higher detection frequency was found for A. actinomycetemcomitans and P. gingivalis for the DNA probe technique; for P. intermedia the detection frequency by culture was higher. For A. actinomycetemcomitans, 21% of the culture positive samples was positive with the DNA probe. Testing the constructed bacterial samples with the DNA probe method resulted in about 16% false positive results for the 3 species tested. Furthermore, 40% of P. gingivalis strains were not detected by the DNA probe. The present data suggest that at least part of the discrepancies found between the DNA probe technique used and cultural methods are caused by false positive and false negative DNA probe results. Therefore, the value of this DNA probe method for the detection of periodontal pathogens is questionable.

Adult↗

The indirect mixed antiglobulin reaction test using a commercially available kit for the detection of antisperm antibodies in serum.

A simple procedure is described for the detection of antisperm antibodies of the IgG class in human serum using the indirect mixed antiglobulin reaction (MAR) test. The test uses only one dilution of serum (1/16) and no washing procedures. The test was performed in 361 sera. Comparative studies were conducted with the tray agglutination test (TAT) in 164 sera and with the direct MAR test on semen in 110 cases. In 24 sera with borderline test results, and in 13 sera with discordant findings, evidence of antisperm antibody activity was investigated by means of additional testing, namely, the adenosine triphosphate release cytotoxicity test (ARCT) and the indirect immunobead tests (IB) for IgA and IgM. Using the value of 40% reaction between motile spermatozoa and coated latex particles as the lower limit of significant activity, the indirect SpermMAR test has a sensitivity of 96% and specificity of 87%. The two false-negative indirect MAR results occurred in cases with IgM, the clinical significance of which is doubtful. Hence, the indirect MAR test appears to be an ideal screening procedure for sperm antibodies in serum since it is easy to perform and sensitive. Considering the false-positive rate of 13%, additional tests should, however, be performed on sera with a reaction of 40% or more in order to assess the relevance of the antibodies as a cause of infertility.

Agglutination Tests↗

[Cavography and CT for demonstration of canalicular spread of renal neoplasms into the inferior vena cava ].

The venous and lymphogenic tumor extension (Robson, Stage III) was assessed in two patient collectives with renal neoplasms: in the first (n = 47), only cavography, in the second (n = 45), only computerized tomography was performed. For both groups there were no false negative diagnoses. False positive readings can occur with CT. Thus, ambiguous findings in CT should be clarified by cavography. CT was superior to cavography regarding the detection of pathologic lymph nodes. However, lymph node enlargement was caused in 10% of these cases by inflammatory hyperplastic reaction without evidence of metastatic tumor spread.

Adolescent↗

Circumoval precipitin (COP) test using air-dried eggs of Schistosoma japonicum and S. mansoni.

In an attempt to establish a simplified circumoval precipitin (COP) test for the diagnostic purpose of schistosomiasis, air-dried eggs of both Schistosoma japonicum and S. mansoni were tested as antigens for this assay. Twenty-six sera from mice infected with S. japonicum showed positive COP reactions as assessed by air-dried eggs. Among 36 serum samples from patients with schistosomiasis japonica, five exhibited false negative reaction when assessed with air-dried eggs and showed a minimum level of COP reaction when assessed with lyophilized eggs. Similarly, all of 30 serum samples from jirds (Meriones unguiculatus) infected with S. mansoni gave positive COP reaction when assessed using air-dried egg. Diagnostic sensitivity of air-dried egg-system was comparable to those of fresh egg-or lyophilized egg-systems. A simple COP technique employing air-dried eggs, instead of lyophilized or fresh ones, would be thus useful for the serodiagnosis of schistosomiasis in local endemic areas, where sophisticated laboratory facilities are not available.

Animals↗

Detection of IgM antibodies directed against the gut-associated circulating cathodic antigen in sera from Schistosoma mansoni infected patients. Development and comparison of three enzyme-linked immunoassays.

The majority of the human IgM antibodies detected with an immunofluorescence assay (IFA) on adult worms are directed against the gut-associated circulating cathodic antigen (CCA). In order to study this phenomenon further we developed and evaluated three related ELISA methods to specifically detect IgM antibodies against purified CCA. The assays employed: 1) direct coating of CCA, 2) indirect coating of CCA via a monoclonal antibody, and 3) IgM antibody-capture by rabbit anti-mu chain antibodies. Using a group of 46 positive sera, it was found that the three ELISA's and the IFA were significantly correlated. To discriminate between positive and negative sera we used a cut-off level of average reactivity + 3 standard deviations of 50 negative sera. False negative reactions were not found in any of the ELISA's, while both in the direct and indirect ELISA one false positive reaction occurred. For further studies or diagnostic use the antibody-capture ELISA is recommended.

Animals↗

Deproteinization of serum: another best approach to eliminate all forms of bilirubin interference on serum creatinine by the kinetic Jaffe reaction.

The negative interference of conjugated, unconjugated, and delta bilirubin on patient serum creatinine determined by the kinetic Jaffe reaction is the unresolved problem. We compared bilirubin interference on thirty patients' serum creatinine obtained from four analyzers, with and without deprotenization before the Jaffe reaction, to the Vitros dry enzymatic method. We found significant negative interference from bilirubin on serum creatinine in all samples directly applied to four wet chemical methods, except the one incorporated with serum blank rate. The negative interferences linearly related to bilirubin concentration. However, bilirubin did not interfere on serum creatinine obtained from all wet chemical methods incorporated with deproteinization process before the reaction. We conclude that deproteinized serum before the reaction is the best approach to eliminate all forms of bilirubin interference on serum creatinine determined by the kinetic Jaffe reaction.

Autoanalysis↗

The popliteal lymph node assay in mice to screen for the immune disregulating potential of chemicals--a preliminary study.

Low mol. wt compounds were tested in the popliteal lymph node (PLN) assay to study whether PLN reactivity could be related to the ability of the compounds to induce autoimmune disorders in man. PLN reactions were measured 7 days after a single subcutaneous (s.c.) injection of dissolved compounds in amounts of 0.3-2.0 mg into one hind footpad of mice and assessed as the weight increase of the draining PLN relative to the PLN weight of the untreated contralateral paw. Hydralazine, chlorpromazine, diphenylhydantoin, carbamazepine, phenylbutazone and nitrofurantoin, all being drugs with a documented potential to induce systemic immunological disorders in man, caused marked PLN reactions. False negative PLN responses were observed following injection of procainamide and isoniazid. Among systemic drugs without known potential to induce autoimmune reactions in humans, quinacrine, denzimol and niridazole significantly increased PLN weights, while phenobarbital, levamisole and disulfiram had no effect. Chemicals with a well-known capacity to induce contact dermatitis in man like 2,4-dinitro-1-chlorobenzene, alpha-methylene-gamma-butyrolactone, p-phenylenediamine, 5-nitro-2-furaldehyde semicarbazone, 2-mercapto-benzothiazol and 1,3-dibutyl-2-thiourea caused marked PLN reactions, while the non-sensitizer 2,4-dichloro-1-nitrobenzene failed to do so. It is concluded that the PLN assay as applied in this study may give a rapid first indication of immunomodulating potential of low mol. wt compounds, but it does not discriminate as to the kind of immunomodulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

First workshop for detection of heparin-induced antibodies: validation of the heparin-induced platelet-activation test (HIPA) in comparison with a PF4/heparin ELISA.

BACKGROUND: No data exist regarding the inter-laboratory reproducibility of the heparin-induced-platelet-activation (HIPA) test, the most widely used functional assay in Germany for the detection of heparin-induced thrombocytopenia (HIT) antibodies. METHODS: Nine laboratories used an identical protocol to test eight different sera with the HIPA test. Five laboratories also tested the sera with a platelet factor 4 (PF4)/heparin-complex ELISA. Cross-reactivity with danaparoid-sodium was assessed using 0.2 aFXa units instead of heparin in the HIPA test. RESULTS: Two of nine laboratories had no discrepant HIPA test results. Four laboratories differed in one sample, one reported two discrepant results, and two laboratories reported more than two discrepant results. Cross-reactivity with danaparoid-sodium test results differed among laboratories. PF4/heparin ELISA results were identical in all five laboratories. CONCLUSION: The HIPA test requires strict quality control measures. Using both a sensitive functional assay (HIPA test) and a PF4/heparin ELISA will allow detection of antibodies directed to antigens other than PF4/heparin complexes as well as detection of IgM and IgA antibodies with PF4/heparin specificity.

Antibodies↗

Comparison of three commercial rapid agglutination test kits for identification of coagulase positive staphylococci from foods and animals.

Three rapid agglutination assays for the identification of Staphylococcus aureus Monostaph (Bionor A/S, Skien, Norway), Staphyslide-Test (BioMerieux, Lyon, France) and Staph-Rapid-Test (Roche, Basel, Switzerland), were compared. A total of 104 Gram-positive, catalase positive cocci were tested: Nineteen Staphylococcus reference strains comprising 15 spp. (4 strains were coagulase positive), and 7 Micrococcus reference strains comprising 4 spp.; 22 food isolates comprising 13 S. aureus, 8 coagulase positive Staphylococcus spp., and 1 Micrococcus sp.; 56 animal isolates comprising 11 S. aureus, 9 S. hyicus subsp. hyicus, 2 S. intermedius, 15 coagulase positive and 19 coagulase negative Staphylococcus spp. Totally 54 strains were coagulase positive. Considering agglutination of a coagulase positive strain as a correct identification, Monostaph, Staph-Rapid-Test, and Staphyslide-Test correctly identified 52 (96.3%), 47 (87.0%) and 48 (89.0%) of the coagulase positive staphylococci, respectively. Monostaph, Staph-Rapid-Test and Staphyslide-Test showed 1 (2.0%), 4 (8.0%) and 4 (8.0%) false positive reactions respectively. Monostaph, Staph-Rapid-Test and Staphyslide-Test gave 0 (0.0%), 6 (5.8%) and 7 (6.7%) non-interpretable reactions, respectively. Monostaph may be a good alternative to the tube-coagulase test for rapid and reliable identification of coagulase positive staphylococci from both food and veterinary sources. However, false negative reactions may occur with coagulase positive strains of S. hyicus subsp. hyicus and S. intermedius.

Agglutination Tests↗

Magnetic resonance imaging in potential postsurgical recurrence of breast cancer: pitfalls and limitations.

OBJECTIVE: To determine the sensitivity and specificity of magnetic resonance imaging (MRI) of the breast for detecting recurrent carcinoma. PATIENTS AND METHODS: Thirteen patients ranging in age from 47 to 77 years who had undergone lumpectomy 5 months to 8 years earlier and who had mammographic findings suggestive of recurrence underwent contrast-enhanced dynamic MRI. Histologic confirmation was obtained in all cases. RESULTS: Of the eight lesions (in seven patients) for which biopsy proved recurrence, MRI correctly identified six; there were two false negative results. Of the six benign lesions, four were correctly identified by MRI. The two false positive results involved fat necrosis and a foreign-body reaction respectively. CONCLUSION: These results confirm previous reports of the poor specificity of MRI of focal breast lesions. The authors therefore recommend caution in the use of breast MRI in the assessment and management of suspected recurrent carcinoma.

Aged↗