The recent introduction of new technologies such as Luminex has provided alternative methods to the Complement Dependent Cytotoxicity (CDC) test for HLA specific antibody detection. In this study we compared the results obtained with CDC to those obtained using a Luminex method with the aim of evaluating the impact of this new technology on antibody screening policies in our transplant setting.A total of 1,421 sera, acquired from patients on the waiting list for a kidney transplant or following transplantation, were tested by both methodologies. CDC was performed using a whole lymphocyte population comprising a panel of 52 cells. The percentage panel reactive antibodies (PRA) and antibody specificity were evaluated using Lambda Scan Analysis software. For the Luminex method sera screening and identification of antibody specificity were carried out using the LABScreen Mixed and LABScreen PRA respectively.The overall concordance between the results obtained using the CDC and the Luminex methods was 85%. HLA antibody specificity was confirmed in 96% of the sera which tested positive using the Luminex system and serum positivity corresponded with a previous sensitisation event in these individuals. Using the Luminex method 18% of patients on the waiting list were considered and managed as sensitised as compared to 7% when testing with CDC alone. The Luminex method was able to detect a number of antibody specificities significantly more frequently than the CDC method and in addition the CDC method failed to detect some of the antibody specificities detected by the Luminex system.Based on this comparison study we have incorporated the Luminex methodology into our screening strategy. q 2007 Clinical Cytometry Society

We performed a prospective study in a cohort of repeat To assess the incidence and source of community-acblood donors to estimate the risk of acquiring HCV infection quired hepatitis C virus (HCV) infection among subjects among individuals without parenteral exposure, and to invesat low risk for blood-borne diseases, we prospectively tigate the possible cause of infection. studied a cohort of 16,515 repeat blood donors over a mean follow-up time of 36 months. Second-and third-PATIENTS AND METHODS generation methods were used for hepatitis C virus antibody (anti-HCV) testing. HCV RNA was determined in A cohort of 18,109 anti-HCV-negative blood donors, candidates the serum of anti-HCV-positive donors by reverse-tran-for a second or subsequent donation, was enrolled between May 1991 scription polymerase chain reaction. Liver biopsy was and April 1992 in a longitudinal prospective study. Of this cohort, 16,515 subjects came to our center for further donations and were performed in the viremic subjects. Risk factors for HCV followed up for an average of 36 months (range, 3-52 months). The infection were identified by a psychosocial questionmedian interval between donations was 202 days. The male/female naire in the whole cohort. During follow-up, 5 donors ratio was 7:3, the median age was 32 years (range, 18-65 years). A became infected with HCV. The incidence was 1 per psychosocial questionnaire, based on 47 direct and indirect ques-10,000 person-years (95% confidence interval, 0.3-2.4 per tions, was routinely administered to blood donors. 16 Questions re-10,000). During the 6 months before seroconversion, four garded: 1) demographic information; 2) cigarette smoking and alcohol subjects (80%) underwent medical or surgical percutane-consumption; 3) drug use, with reference to type, frequency, and time ous procedures, compared with 26.5% in the entire donor of last assumption; 4) sexual history; 5) history of hepatitis; and 6) cohort (difference between frequencies, 53.5%; CI: 18.9-other risk factors for blood-borne infection (blood transfusion, tattooing, nosocomial exposure). At each donation, the clinical data The subjects showing confirmed anti-HCV reactivity were asked after seroconversion, and liver biopsy showed chronic to enter a follow-up program consisting of clinical and laboratory hepatitis in all cases. Thus, new cases of hepatitis C oc-evaluation, including HCV-RNA determination, at 1-to 3-month incur among individuals without a history of known risk tervals. A serum sample, collected on the blood donation preceding factors, some of which may be caused by nosocomial seroconversion, was retested for anti-HCV using a third-generation exposure. (HEPATOLOGY 1997;25:702-704.) test and was evaluated for HCV RNA. Liver biopsy was proposed to the viremic subjects. Sexual partners and household contacts of seroconverting subjects were invited for counseling and anti-HCV Chronic hepatitis C virus (HCV) infection represents a sig-testing. nificant cause of morbidity and mortality worldwide. 89.1). One seroconverti...

We investigated the iron status of 33 pyruvate kinase (PK) deficient patients, most of the cases reported in Italy. Serum ferritin (SF) was higher than the upper limit of the range of matched controls in 15/25 (60%) non-transfused patients (median 228 micrograms/l, range 58-3160 v 43, 22-310). Liver siderosis and fibrosis were found in 8/9, and cirrhosis in two who died at age 39 and 42 of complications of iron overload. SF was independent of age, sex, or severity of haemolysis. The prevalence of HLA-A3 antigen in PK deficient patients was not significantly different from that of our healthy population (29.6% v 23%). The HLA-A3 positive, non-transfused patients had significantly higher SF values than the HLA-A3 negative ones (median 675 micrograms/l, range 340-3160 v 145, 58-400). A pedigree study of six high SF-probands indicated that iron overload has a multifactorial pathogenesis. In particular, the association of PK deficiency-induced haemolysis, splenectomy and an additional factor (heterozygosity for idiopathic haemochromatosis, ineffective erythropoiesis) leads to severe iron accumulation. We suggest that monitoring iron status would be useful in PK deficient patients, particularly in splenectomized and HLA-A3 positive ones, to identify those at risk of iron overload and prevent the clinical consequences of iron accumulation.

Eleven new cases of red cell pyruvate kinase (PK) deficiency with congenital haemolytic disease from 10 unrelated Italian families were characterized using the methods recommended by the International Committee for Standardization in Haematology (ICSH). All patients were double heterozygotes for the PK gene. The 10 variants were designated PK 'Lecce,' 'Parma,' 'Verona,' 'Milano,' 'Soresina,' 'Macerata,' 'Sassari,' 'Genova,' 'Mantova' and 'Brescia.' PK 'Sassari' was associated with glucose-6-phosphate dehydrogenase deficiency in two siblings. All mutants displayed multiple biochemical abnormalities except for PK 'Lecce' that only showed decreased red cell PK activity. No relation was found between the severity of anaemia and either the residual PK activity or specific biochemical enzyme abnormalities. Increased serum ferritin levels were detected in most of the patients, suggesting the need for systematically monitoring iron status in this disease.

In some pathological conditions, HLA class II antibodies can react with activated granulocytes expressing HLA-DR antigens, and activate TRALI reaction. HLA class II antibodies screening and flow cytometry cross-matching techniques should be added to the current diagnostic algorithm of TRALI.