Multicentre evaluation of the new ORTHO VISION® analyser.

BACKGROUND: Implementation of fully automated analysers has become a crucial security step in the blood bank; it reduces human errors, allows standardisation and improves turnaround time (TAT). OBJECTIVES: We aimed at evaluating the ease of use and the efficiency of the ORTHO VISION® Analyser (VISION) in comparison to the ORTHO AutoVue® Innova System (AutoVue) in six different laboratories. METHODS: After initial training and system configuration, VISION was used in parallel to AutoVue following the daily workload, both automates being based on ORTHO BioVue® System column agglutination technology. Each participating laboratory provided data and scored the training, system configuration, quality control, maintenance and system efficiency. A total of 1049 individual samples were run: 266 forward and reverse grouping and antibody screens with 10 urgent samples, 473 ABD forward grouping and antibody screens with 22 urgent samples, 160 ABD forward grouping, 42 antibody screens and a series of 108 specific case profiles. RESULTS: The VISION instrument was more rapid than the AutoVue with a mean performing test time of 27·9 min compared to 36 min; for various test type comparisons, the TAT data obtained from VISION was shorter than that from AutoVue. Moreover, VISION analysed urgent STAT samples faster. Regarding the ease of use, VISION was intuitive and user friendly. CONCLUSIONS: VISION is a robust, reproducible system performing the most types of analytical determinations needed for pre-transfusion testing today, thus accommodating a wide range of clinical needs. VISION brings appreciated new features that could further secure blood transfusions.

Haemostatic function and biomarkers of endothelial damage before and after platelet transfusion in patients with acute myeloid leukaemia.

OBJECTIVES: The beneficial effect of platelet transfusion on haemostasis is well established, but there is emerging evidence that platelet transfusion induces an inflammatory response in vascular endothelial cells. BACKGROUND: We investigated haemostatic function and endothelial biomarkers before and after platelet transfusion in patients with acute myeloid leukaemia. MATERIALS AND METHODS: Blood was sampled before, 1 and 24 h after platelet transfusion. Primary and secondary haemostasis was evaluated by whole blood aggregometry (Multiplate) and thromboelastography (TEG). Endothelial biomarkers (sICAM-1, syndecan-1, sThrombomodulin, sVE-Cadherin) and platelet activation biomarkers (sCD40L, TGF-beta) were investigated along with haematology/biochemistry analyses. RESULTS: Twenty-two patients were included. Despite continued low platelet counts, platelet transfusion normalised the median values of most TEG parameters and slightly increased platelet aggregation (all P < 0·05). Endothelial biomarkers were not significantly affected by transfusion. The 1 h sCD40L level correlated positively with Syndecan-1 and soluble thrombomodulin delta values, biomarkers of endothelial damage (both P = 0·005). CONCLUSION: Platelet transfusion improved haemostasis, whereas post-transfusion increases in sCD40L were associated with endothelial damage, indicating that transfused platelets and platelet-derived pro-inflammatory mediators may have opposite effects on the endothelium.

[Occurrence of hepatitis B and C among mentally retarded]. / Forekomst af hepatitis B og C blandt mentalt retarderede.

The objective of this study was to examine whether the prevalence of hepatitis C, like hepatitis B, is increased among the mentally retarded in Denmark. The prevalence of serological markers of hepatitis B and C was examined in an institution for the mentally retarded. A total of 126 out of 178 inhabitants (71%) with a median age of 49 years (range 23-78) participated. All subjects were anti-HCV-negative by third generation ELISA antibody test. A total of 45 (35.7%) subjects were anti-HBc-positive and 10 (7.9%) were HBsAg-positive. Among subjects with Down's syndrome (n = 20), 55% were anti-HBc-positive and 30% were HBsAg-positive as compared to 32% and 3.8% respectively among others. In conclusion, hepatitis C infection seems to be uncommon among mentally retarded persons in Denmark and the risk of acquiring infection not significantly increased as compared to that of the general population. The prevalence of serological markers for hepatitis B was high and comparable to previous studies in this population.

Alpha interferon therapy in Danish haemophiliac patients with chronic hepatitis C: results of a randomized controlled open label study comparing two different maintenance regimens following standard interferon-alpha-2b treatment.

Following a survey among all Danish haemophiliac patients 49 HIV-negative patients with chronic hepatitis C were offered enrollment in a randomized controlled open label study comparing two different maintenance regimens following standard interferon-alpha-2b treatment. Dose modifications and treatment discontinuation were based upon changes in transaminase levels. Forty-seven patients enrolled received 3 MU of alpha interferon thrice weekly (TIW) for 3 months. Twenty-six nonresponders had their dose increased to 6 MU TIW for an additional 3 months, while 21 responding patients continued on 3 MU TIW. At 6 months, 25 patients with a complete or a partial biochemical response were randomly allocated to either a fixed dose regimen (13 patients) (3 or 6 MU thrice weekly) or an individualized dose regimen (12 patients) tapering interferon dose from 3 or 6 MU by one-third every 2 months if transaminases were persistently normal. The remaining 22 biochemical nonresponders were followed for an additional 6 months without further treatment. After 12 months of treatment, 18 patients (38%) had a virological response, irrespective of regimen, and seven patients (16%) had a sustained virological and biochemical response after 6 months of follow up. Overall, the individualized treatment regimen did not seem to offer any advantage over the fixed dose regimen. The response to alpha interferon treatment in Danish haemophiliac patients with chronic hepatitis C immediately after treatment is comparable to that obtained in previous studies among nonhaemophiliacs. However, a sustained virological and biochemical response was seen in only 16% of treatment patients.

The seroprevalence of hepatitis B and C in hospitalized Danish patients.

Health care workers are at risk of acquiring blood-borne infections. To assess the risk of exposure to hepatitis B or C in the case of occupational blood exposure, we determined the seroprevalence of these infections in 466 patients admitted to a Copenhagen university hospital. Serological markers for hepatitis B or C were detected in 56 patients (12.0%). The seroprevalence of HBsAg and anti-HCV was 0.9% and 1.5% respectively. HCV RNA, indicating ongoing hepatitis C, was found in five of seven anti-HCV-positive patients by polymerase chain reaction. The serological findings had not previously been diagnosed in 4 of 10 potentially infectious patients and only 6 of 10 patients belonged to high-risk groups. In conclusion, health care workers should be aware of the potential the occupational risk of hepatitis B and C even in a low-prevalence country like Denmark. Management of health care workers after blood exposure should include serological testing for both hepatitis B and C. Strict adherence to universal precautions is recommended and vaccination against hepatitis B should be encouraged.

High prevalence of hepatitis C virus (HCV) RNA in dialysis patients: failure of commercially available antibody tests to identify a significant number of patients with HCV infection. Copenhagen Dialysis HCV Study Group.

Results of serologic tests were correlated with hepatitis C virus (HCV) viremia, determined by a cDNA polymerase chain reaction assay to detect HCV RNA, in 340 Danish dialysis patients; of these, 28 (8.2%) were positive for antibodies to HCV (anti-HCV) with second-generation ELI-SAs. HCV RNA was found in sera from 27 of these 28 anti-HCV-positive patients. However, 8 dialysis patients had detectable levels of HCV RNA but were anti-HCV-negative with second-generation ELISAs. Among the 35 HCV-infected dialysis patients 16 were positive, 7 indeterminate, and 12 negative with the second-generation RIBA. More than 60% of patients with evidence of ongoing liver disease had HCV infection. Thus, current commercially available antibody tests did not accurately reflect the HCV status in dialysis patients. A relatively high prevalence (> 10%) of HCV RNA, closely associated with liver disease, was found among dialysis patients in a low-prevalence area of the world.

Drug metabolism and genetic polymorphism in subjects with previous halothane hepatitis.

To test the hypothesis that halothane hepatitis is caused by a combination of altered drug metabolism and an immunoallergic disposition, the metabolism of antipyrine, metronidazole, sparteine, phenytoin, and racemic R- and S-mephenytoin was investigated in seven subjects with previous halothane hepatitis. The HLA tissue types and the complement C3 phenotypes were also determined. The metabolism of antipyrine and metronidazole was within normal range in all subjects, and they were all fast or extensive metabolizers of sparteine, mephenytoin, and phenytoin. HLA tissue types were unremarkable. Five of the seven subjects had complement C3 phenotypes F or FS. In the general population phenotype S is the most common, but the difference in complement C3 phenotypes is not statistically significant (p = 0.07). We conclude, although in a limited number of patients, that subjects with previous halothane hepatitis do not appear to be different from controls with regard to drug metabolism and HLA tissue type. The possibility of a higher frequency of complement C3 phenotype F and FS needs further investigation.

Hepatitis C in dialysis patients: relationship to blood transfusions, dialysis and liver disease.

Antibodies to hepatitis C virus (anti-HCV) were determined in an unselected group of 340 patients with chronic renal failure treated with maintenance dialysis. A second generation hepatitis C virus (HCV) enzyme-linked immunosorbent assay (ELISA) was used and confirmation made by a second generation recombinant immunoblot assay (RIBA). Sixteen patients (4.7%) were anti-HCV positive and 8 (2.4%) were anti-HCV indeterminate. All anti-HCV positive and anti-HCV indeterminate patients had received blood transfusions. No statistically significant differences were found between anti-HCV positive and indeterminate patients considering blood transfusions, dialysis and liver disease. The combined group of anti-HCV positive and indeterminate patients had had more blood transfusions (P < 0.005) and had been on dialysis for a longer period (P < 0.01) compared with anti-HCV negative patients. Further, significant correlation with elevation of transaminases and anti-HCV was observed (P < 0.001). Thirty patients (8.8%) had elevated transaminase levels and 13 (43%) of these were anti-HCV positive or indeterminate. In conclusion, HCV infection accounts for a substantial proportion of liver disease in dialysis patients, probably most often transmitted by blood transfusions but other routes of transmission could not be excluded.

Hepatitis C virus antibodies in homosexual men and intravenous drug users in Denmark.

In order to evaluate the role of sexual transmission and parenteral transmission of hepatitis C virus (HCV) in homosexual men and intravenous drug users (IVDU) serum samples from 147 homosexual men and 126 IVDU were tested for anti-HCV. Anti-HCV was found in two (1.4%) of the homosexual men and in 123 (98%) of IVDU. The presence of anti-HCV could not be correlated to the presence of HBV markers or HIV-antibodies. HCV is widespread among Danish IVDU. Risk of sexual transmission seems low even though sexual contact is a much more prevalent risk factor than needle sharing.

Clinical manifestations in patients with autoantibodies specific for nuclear lamin proteins.

IgG antibodies to nuclear lamin proteins have been found in serum samples from 31 patients using immunofluorescence on HEp-2 cells, Western blotting, and enzyme-linked immunosorbent assay, performed against a nuclear lamina preparation from Ehrlich ascites tumor cells. Antilamin antibodies were most prevalent among patients with nonerosive, seronegative polyarthritis, or patients showing serum antiphospholipid reactivity as well. It is possible that anti-lamin antibodies may thus be a marker for a subgroup of polyarthritis patients who have a different prognosis from that of those with seropositive rheumatoid arthritis. The mechanism for the combined occurrence of anti-lamin and antiphospholipid autoantibodies is obscure. Future studies will answer whether these two antibodies represent a distinct antibody profile in patients with antiphospholipid antibody syndrome.

Chronic evolution of acute hepatitis B: the significance of simultaneous infections with hepatitis C and D. Copenhagen Hepatitis Acuta Programme.

Seventy-six of 77 consecutive patients with hepatitis B surface antigen (HBsAg)-positive acute hepatitis were reevaluated using anti-hepatitis C virus (HCV), anti-hepatitis D virus (HDV), and IgM anti-hepatitis B core (HBc) testing. Anti-HCV and/or anti-HDV was found in 32 patients (42%). The presence of these markers was significantly associated with intravenous drug abuse (p less than 10(-6). Sixty-nine patients were IgM anti-HBc-positive, of whom two (3%) (95% confidence limits, 1-12%) became chronic HBsAg carriers with histologically verified chronic liver disease; both were anti-HCV and anti-HDV-negative. Among the remaining 67 IgM anti-HBc-positive patients 8 had HBV and HDV co-infection, 3 had HBV and HCV co-infection, and 1 had HBV, HCV, and HDV co-infection. Twenty-two had evidence of preceding or past HCV infection; two developed chronic active hepatitis in spite of HBsAg clearance. Seven patients with IgM anti-HBc negative. One was a chronic HBsAg carrier with HDV superinfection. One had subclinical acute HBV infection and became a chronic HBsAg carrier. In a further two patients reactivation of replication in a chronic HBV infection could not be disregarded. Three patients could not be classified; all had acute recent onset of symptoms, cleared HBsAg within 6 months, but lacked IgM anti-HBc. It is concluded that HCV and HDV superinfections in HBV carriers mimicking acute HBV infection with chronic evolution are rarely encountered in the present population in spite of high frequency of both HCV and HDV markers.

[Screening of Danish blood donors for hepatitis C virus antibodies]. / Screening af danske bloddonorer for antistof rettet mod Hepatitis C virus.

In Denmark only 1-3 cases of transfusion-associated hepatitis NANB (TAH-NANB) are registered annually or about 1 case per 100,000 units transfused. Thus, the prevalence of blood donors with hepatitis NANB infection is estimated to be very low and measures to prevent TAH-NANB such as surrogate testing have not been considered to be worth while. The incidence of TAH-NANB, however, may be underestimated if cases are overlooked or not identified as transfusion-associated or if registration fails. The appearance of the anti-HCV test (Ortho) has clearly provided a more sensitive and specific tool for identifying blood donors, who may transmit hepatitis NANB. In the present study, we have determined the prevalence of anti-HCV in random Danish blood donors from 1989 (fresh serum samples) and 1982 (frozen serum samples). All donors were tested for ALT and anti-HBc. A prevalence of 0.3% anti-HCV positive blood donors was found in 1989 (2/600) as well as in 1982 (2/595). Thus, no change in the prevalence of HCV infected donors seems to have occurred in Denmark during these years. The anti-HCV prevalence of 0.3% in random blood donors is low but not negligible, and if the majority of the reactive donors identified are true positives, the epidemiologic pressure as to HCV infection probably equalizes the pressure as to HBV infection (0.1-0.2%) in people attending our blood banks to become donors. In a group of donors positive for surrogate markers (ALT greater than 40 U/l and/or anti-HBc) we found only 0.9% (2/225) anti-HCV positives.(ABSTRACT TRUNCATED AT 250 WORDS)

Sexual transmission of hepatitis C virus: cohort study (1981-9) among European homosexual men.

OBJECTIVE: To determine the prevalence, incidence, and persistence of positivity for antibodies to hepatitis C virus (anti-HCV) and the potential for sexual transmission of the virus. DESIGN: A cohort analysis covering 1981-9 comparing estimated cumulative incidences of and seroconversion rates for anti-HCV with those of hepatitis B core antibody (anti-HBc) and antibodies to the human immunodeficiency virus (anti-HIV). SETTING: Copenhagen and Aarhus, Denmark. SUBJECTS: 259 Male members of a Danish homosexual organisation. MAIN OUTCOME MEASURES: Correlations of prevalence and incidence with a wide range of sexual lifestyle variables. RESULTS: Only four (1.6%) subjects were positive for anti-HCV in 1981. The estimated cumulative incidence of positivity for anti-HCV was 4.1% in 1984 (seroconversion rate during 1981-4 (2.5%)) and remained at 4.1% in 1989 (seroconversion rate nil during 1984-9). In contrast, positivity for anti-HBC rose from 44.0% in 1981 to 52.7% in 1984 (seroconversion rate 15.5%) and 58.8% in 1989 (seroconversion rate 12.9%), and that for anti-HIV rose from 8.8% to 24.0% (seroconversion rate 16.7%) and 30.1% (seroconversion rate 8.0%) respectively. Three anti-HCV positive patients seroreverted three to five years later. None of the anti-HCV positive subjects had had a transfusion and only one gave a past history of intravenous drug use. Variables in sexual lifestyle correlated with the presence of anti-HBc but not with that of anti-HCV. CONCLUSIONS: In contrast with hepatitis B virus and HIV, sexual transmission of hepatitis C virus seems to be a rare event. Furthermore, antibodies to the virus may become undetectable after several years.

Early appearance of antibodies to hepatitis C virus in community acquired acute non-A, non-B hepatitis is associated with progression to chronic liver disease. The Copenhagen Hepatitis Acuta Programme.

24 consecutive patients (14 females; median age 36, range 18-77) with liver biopsy proven acute non-A, non-B hepatitis (NANBH) were assayed for antibodies to hepatitis C virus (HCV). 14 (58%) were positive initially or during follow-up. Three patients were positive within 4 weeks following onset of symptoms and 7 patients in a serum sample obtained 4-8 weeks after clinical onset. Seroconversion was documented in 7/8 patients in paired sera from the acute phase of the disease. Anti-HCV was detected in 6% and 13% of control patients with acute hepatitis A and toxic hepatitis. NANBH in 6/14 patients (43%) with anti-HCV progressed to chronic liver disease (CLD). In contrast none of the anti-HCV negative patients developed CLD (p = 0.02). In addition, 2 anti-HCV positive patients developed fulminant and fatal hepatitis. The predominant route of HCV transmission was intravenous drug abuse. It is concluded that hepatitis may be ascribed to HCV infection in more than half of patients with community aquired NANBH, that seroconversion occurs in the majority within 8 weeks following onset of symptoms and that seropositive individuals often progress to CLD.

No effect of oral testosterone treatment on sexual dysfunction in alcoholic cirrhotic men.

The prevalence and course of sexual dysfunction was evaluated in 221 alcoholic cirrhotic men participating in a double-blind, placebo-controlled study on the effect of oral testosterone treatment on liver disease. At entry, 67% (95% confidence limits, 61%-74%) complained of sexual dysfunction. Sexual dysfunction was significantly (p less than 0.05) associated with lower serum concentrations of testosterone, non-protein-bound testosterone, and non-sex hormone-binding globulin-bound testosterone. The significant associations between sexual dysfunction and non-protein-bound and non-sex hormone-binding globulin-bound testosterone concentrations disappeared, however, when age, ethanol consumption, and severity of liver disease were included as covariates in the analysis. During follow-up (median 30 mo, range 1-48 mo) sexual dysfunction improved significantly (p less than 0.05) at 6, 12, and 24 mo. Furthermore, the reported libido and erectile and ejaculatory function improved significantly at the end of the follow-up period (p less than 0.01). However, the testosterone-treated patients did not differ significantly from the placebo-treated patients regarding any of the changes in sexual function. In conclusion, oral testosterone treatment does not significantly influence the type or course of sexual dysfunction in alcoholic cirrhotic men. However, sexual function improved after reduction of ethanol consumption in these patients.