Liver Disease and HPLC Quantification of Disialotransferrin for Heavy Alcohol Use: A Case Series

Abstract

Background It had previously been suggested that individuals with cirrhosis may have a pattern of transferrin glycosylation that interferes with the interpretation of carbohydrate-deficient transferrin (CDT) testing for heavy alcohol use. The goal of this case series was to evaluate the prevalence of liver disease among individuals with poor resolution of transferrin glycoforms by high performance liquid chromatography. Methods We reviewed the electronic medical records of 35 consecutive patients with poor chromatographic resolution of disialotransferrin from trisialotransferrin, and recorded information on diagnosed liver disease, liver function testing, and other factors. Results Thirty of the 35 subjects with poor chromatographic resolution of the transferrin glycoforms had sufficient data in the medical record for some estimation of liver function. Of these 30 subjects, 25 had previously diagnosed liver pathology. Of the remaining 5 subjects, 2 had liver imaging results suggestive of benign tumor; the remaining 3 had mildly elevated bilirubin and aminotransferase activity, and low albumin. Conclusions Liver abnormalities, but not necessarily cirrhosis, are common in individuals with poor chromatographic separation of transferrin glycoforms, which might lead to false positive results on CDT testing. However, the chromatographic-based assay can detect this issue, minimizing the reporting of false positives, but not necessarily assisting in valid detection of heavy drinking.

Relationship of Abnormal Chromatographic Pattern for Carbohydrate-Deficient Transferrin with Severe Liver Disease

“…Di-tri bridging was originally observed in the laboratory without detailed clinical data, and was hypothesized to represent a genetic variant (Helander et al, 2001). Although genetic variation could contribute to this pattern, in subsequent case reports it has been associated with significant liver disease (Arndt et al, 2008;Stewart et al, 2010;Gonzalo et al, 2012). The current study confirmed and extended these observations.…”

“…Despite its utility in identifying chronic heavy drinking, CDT assay methods have diminished specificity for heavy alcohol use in patients with liver disease and particularly cirrhosis (Heinemann et al, 1998;DiMartini et al, 2001). Case series using the relatively newer HPLC or capillary electrophoresis assays for %dCDT have shown that liver disease can be associated with a diminished chromatographic resolution of disialotransferrin from trisialotransferrin (Arndt et al, 2008;Stewart et al, 2010;Gonzalo et al, 2012), which is the likely cause of this diminished specificity for heavy drinking. This abnormal pattern is due to the presence of higher mass disialotransferrin isoforms resulting from liver-diseaseassociated changes in transferrin glycosylation (Landberg et al, 2012).…”

BMI But Not Stage or Etiology of Nonalcoholic Liver Disease Affects the Diagnostic Utility of Carbohydrate‐Deficient Transferrin

“…Previously, it has been reported that stage or etiology of liver disease may affect %CDT (Arndt et al, 2008;Fleming et al, 2004;Stewart et al, 2010). In our patient cohort, no significant difference in %CDT was noted in relation to gender, smoking, age, or ethnicity.…”

“…HPLC provides a graphic visualization of the individual transferrin glycoforms permitting the detection of factors that interfere with the analysis. Visible assessment of glycoform patterns may be important in patients with liver disease because cirrhosis has been reported to lead to poor chromatographic resolution of disialotransferrin from trisialotransferrin (di-tri bridging), a pattern that could result in inaccurate quantification (Arndt et al, 2008;Stewart et al, 2010). Prior to the introduction of the standardized method, it was reported that CDT was elevated in patients with liver disease in the absence of documented alcohol abuse (Fleming et al, 2004).…”

Disialo–trisialo bridging of transferrin is due to increased branching and fucosylation of the carbohydrate moiety

“…It was recently established that the di-tri bridging phenomenon could neither be explained by a genetic transferrin protein variant nor by an increased trisialotransferrin fraction [16]. It has been suggested that the di-tri bridging phenomenon may be prevalent in patients with liver cirrhosis or other severe liver abnormalities [16,17].…”