Clinical utility of liquid-chromatographic analysis of effusions for hyaluronate content

Abstract

A previously described HPLC method for determining hyaluronate in effusions was used to analyze a consecutive series of effusions from 1039 patients with pleural fluids and from 571 patients with peritoneal fluids. A mesothelioma was verified histologically in 50 of the cases. The results were used to estimate the clinical utility of the analysis. With a cutoff of 75 mg/L for hyaluronate-derived uronic acid, assay specificity for a malignant mesothelioma was 100% and the sensitivity 56%. Only 20% of the effusions from the mesothelioma patients showed no evidence of increased production of hyaluronate. Cytological smears from the associated cell pellets were evaluated as malignant or suspicious for malignancy in only 28% or in a further 46% of the mesothelioma cases, respectively, leaving 30% of the pellets as cytologically false-negative. We also analyzed effusions from selected cases submitted from other hospitals, 154 of which had been diagnosed histologically as mesotheliomas. Concentrations of hyaluronate were increased in these cases too, but a considerable proportion of the samples showed evidence of losses of hyaluronate; consequently, the sensitivity of the assay in these samples was lower.

Telomerase activity analyzed with TRAP in situ provides additional information in effusions remaining equivocal after immunocytochemistry and hyaluronan analysis

“…Effusions represent a major diagnostic challenge in clinical cytology and a combination of microscopic morphological evaluation and immunocytochemical staining is often necessary for an adequate diagnosis. [1][2][3] In our laboratory, we also use hyaluronan content to distinguish mesotheliomas from metastatic tumors and from benign mesothelial cells, 3,4 but around 10% of the effusions still remain unsolved, which is in agreement with other studies. 5,6 The spectrum of underlying diseases in the diagnostically refractory effusions differs considerably from those in unselected effusions and the samples may even lack diagnostic cells.…”

The combination of CEA, EMA, and BerEp4 and hyaluronan analysis specifically identifies 79% of all histologically verified mesotheliomas causing an effusion

“…The peaks obtained at 231 nm were recorded and compared with the peaks obtained for external standards. 16 In a previous study, we have shown that values exceeding 75 mg/l of hyaluronan-derived uronic acid are specific for a mesothelioma. The probability of a mesothelioma is highly increased also in effusions with values higher than 25 mg/l but the specificity then is reduced to 95%.…”

“…14,15 A high-performance liquid chromatography (HPLC)-based method developed by Hjerpe and coworkers has proved to be specific enough and sensitive enough for routine diagnostic purposes, also in long-time use. 16 This analysis has the potential to identify a mesothelioma also in those cases where no diagnostic cells are shed into the effusion.…”

Differentiation of mesothelioma from adenocarcinoma in serous effusions: The role of hyaluronic acid and CD44 localization

“…Our findings supported the notion that the concentration of HA in malignant pleural effusion caused by MM is significantly higher than that in malignant pleural effusions associated with ACAs. 31,32 In many early studies on cell-cell aggregation, HA was found to mediate cross-linking of cells. Such cross-bridging suggests the existence of membrane-localized HA binding sites.…”