Limits of usual biochemical alcohol markers in cord blood at term: a fetal/maternal population-based study.

BACKGROUND: After maternal alcohol consumption during pregnancy, many neonates affected by less apparent forms of fetal alcohol syndrome disorder (FASD) do not receive proper diagnosis or treatment. There is thus a need for laboratory markers for early detection of alcohol-exposed neonates. The aim of our study was to assess the efficiency of the usual alcohol biomarkers measured in cord blood to identify alcohol-exposed neonates immediately after birth. METHODS: A 1-year study was conducted in the labor wards of the maternity units of the Auvergne, Central France at the time of term delivery. The patients answered an anonymous self-completion survey concerning alcohol use (Alcohol Use Disorder Identification Test; AUDIT) during their pregnancy. Aspartate aminotransferase, alanine aminotransferase, and gamma-glutamyltransferase concentrations and the percentage of carbohydrate-deficient transferrin were measured in maternal and cord blood. RESULTS: We collected 870 maternal-fetal sample pairs. Two cases (0.2%) of typical FASD were detected. We report a non-significant correlation between maternal and cord blood biomarkers. None of the cord blood biomarkers differed significantly between newborns of alcohol-exposed and unexposed mothers. CONCLUSIONS: We demonstrate that the usual alcohol biomarkers are not effective in cord blood for identifying alcohol-exposed neonates.

Performance of asialotransferrin in detecting alcohol abuse.

BACKGROUND: The spectrum of alcohol use disorders covers hazardous use, alcohol abuse, and alcohol dependence. The present study evaluated the performance of asialotransferrin, a newly proposed biomarker for alcohol use disorders, in detecting alcohol abuse and alcohol dependence. METHOD: A 4-month trial was conducted in three groups of participants: alcohol abusers and alcohol-dependent patients, as defined in DSM-IV, and a control group. Asialotransferrin was assayed by capillary zone electrophoresis. RESULTS: Asialotransferrin demonstrated a sensitivity of 0.34 and a specificity of 1.00 for alcohol abuse. The sensitivity of asialotransferrin increased to 0.57 in alcohol-dependent patients. CONCLUSION: Despite the high specificity of asialotransferrin in alcohol use disorders, its sensitivity is too low to make it a useful marker of alcohol abuse.

Analyte comigrating with trisialotransferrin during capillary zone electrophoresis of sera from patients with cancer.

BACKGROUND: Serum concentrations of monoglycosylated isoforms of transferrin are increased by chronic ethanol intake. We investigated transferrin glycosylation in patients with cancer, in which aberrant glycosylation is also induced. METHODS: We used a P/ACE 5000 series capillary zone electrophoresis (CZE) apparatus and a CZE carbohydrate-deficient transferrin reagent set to study 200 cancer patients who consumed alcohol moderately and 33 who were alcohol abusers; we then compared these patients with 56 healthy teetotalers, 89 moderate, and 112 excessive alcohol drinkers without known malignancies. Transferrin isoforms were identified by immunosubtraction with anti-human transferrin polyclonal antibody. RESULTS: Seven peaks, P0-P6, were visualized and completely or partly immunosubtracted when CZE separation was performed at pH 8.5. P0 was present in 95% of alcohol abusers with or without cancer. P3 was significantly higher in cancer patients and was only partly immunosubtracted as trisialotransferrin in all groups. The comigrating analyte was not altered by papain, precipitation by ethanol, or extraction by organic solvents, but was sensitive to acid hydrolysis, suggesting a polysaccharide structure. When isolated at pH 8.25, this analyte was higher in cancer patients. ROC curve analysis identified localized malignant neoplasia at P3 values above 5.8% of total transferrin (sensitivity, 0.78; specificity, 0.87), regardless of alcohol consumption. Disseminated cancers were better differentiated above 8% (sensitivity, 0.94; specificity, 0.96). CONCLUSIONS: Malignant neoplasia, unlike excessive ethanol intake, did not alter the addition of two N-glycans to transferrin. A peak comigrating with trisialotransferrin had characteristics of a polysaccharide in all adults and was increased in sera of patients with cancer.

Use of capillary zone electrophoresis for differentiating excessive from moderate alcohol consumption.

BACKGROUND: The poorly sialylated transferrin isoforms in serum were analyzed by capillary zone electrophoresis (CZE) to differentiate moderate from heavy alcohol consumption. METHODS: We enrolled 614 volunteers, classified after interviews, self-reported drinking habits, and AUDIT scores as alcohol abusers (consuming >50 g/day ethanol for the previous 3 months or longer; n = 413) or moderate drinkers (<30 g/day ethanol; n = 201). Serum transferrin isoforms were separated at 28 kV and monitored at 214 nm on a P/ACE 5500 CZE with use of fused-silica capillaries and the related CEofix CDT reagent set. Immunosubtraction by anti-human transferrin and electrophoretic migration times identified the isoforms. Previous markers of alcohol abuse and an assay combining anion-exchange minicolumn chromatography with immunoturbidimetry (%CDT) were included in the study. Sensitivities and specificities were compared by ROC analysis. RESULTS: The asialylated isoform was missing in 95% of moderate drinkers but present in 92% of alcohol misusers. Disialotransferrin had a specificity and sensitivity of 0.75 at a cutoff of 0.7% of total transferrin, whereas the sum (asialo- + disialotransferrin) at a threshold of 1.2% had a sensitivity of 0.73 and a specificity of 0.92. Trisialotransferrin values did not distinguish between the two populations. Sensitivities and specificities of %CDT averaged 0.77 and 0.74, respectively, at a 2.6% cutoff; 0.67 and 0.83 at 2.8%; and 0.63 and 0.90 at 3%. CDT data were more sensitive and specific for males. Conventional biomarkers appeared less discriminating. CONCLUSIONS: Asialotransferrin detected by CZE in sera of alcohol abusers offers the highest discrimination between excessive and moderate drinking.

Carbohydrate-deficient transferrin isoforms measured by capillary zone electrophoresis for detection of alcohol abuse.

BACKGROUND: Measurements of carbohydrate-deficient transferrin (CDT) are used as markers of alcohol abuse. We developed a capillary zone electrophoresis (CZE) method aimed at improving accuracy of CDT testing. METHODS: We studied 111 alcohol abusers with Alcohol Use Disorders Identification Test scores >11 and 50 teetotalers. CZE was performed with a P/ACE 5500, fused-silica capillaries, and a CEofix CDT reagent set. After iron saturation, sera were loaded by low-pressure injection, separated at 28 kV, and monitored at 214 nm. We identified the transferrin isoforms by migration times, treatment with 100 U/L neuraminidase, and immunosubtraction with anti-human transferrin and anti-C-reactive protein antibodies. We compared CZE results with current biological markers of alcohol abuse, including the %CDT turbidimetric immunoassay. RESULTS: Migration times of the isoforms were identical in both populations. Asialotransferrin was missing in teetotalers but present in 92% of alcohol abusers. Disialotransferrin was higher in those who consumed excessive amounts of alcohol, whereas mean trisialotransferrin concentration was not affected by alcohol abuse. At cutoffs to maximize sensitivity and specificity, these values were 0.92 and 1 [mean ROC area (MRa), 0.96; 95% confidence interval (CI), 0.93-0.99] for asialotransferrin; 0.84 and 0.94 for the sum of asialo- + disialotransferrin (MRa, 0.94; 95% CI, 0.91-0.98); 0.79 and 0.94 for disialotransferrin (MRa, 0.89; 95% CI, 0.84-0.94); 0.62 and 0.53 for trisialotransferrin (MRa, 0.58; 95% CI, 0.49-0.68); 0.79 and 0.82 for a 3% %CDT; and 0.83 and 0.69 for a 2.6% cutoff (MRa, 0.87; 95% CI, 0.81-0.92). Current markers lack sensitivity (<0.65). Transferrins were not significantly correlated with serum enzymes and mean erythrocyte volume. CONCLUSIONS: CZE-isolated desialylated transferrin isoforms allowed differentiation between chronic alcohol abusers and teetotalers.