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Objectives: Increased frequency of hyperamylasemia has previously been reported in human immunodeficiency virus (HIV)-positive patients, but studies determined total amylase activity and were performed before the introduction of highly active antiretroviral therapy (HAART). We evaluated the frequency of pancreatic hyperamylasemia in a large HIV+ population mostly treated with HAART.Methods: The upper reference limit (URL) for pancreatic amylase (P-AMY) was derived from 299 healthy blood donors. A cross-sectional study was then performed on samples obtained from 1,548 consecutive patients referred to our infectious disease clinic to assess serum P-AMY and lipase concentrations. Of the patients, 94% were HIV+, and most (92%) were taking HAART (HIV+Tx+).Results: P-AMY URL was 51 U/L. The frequency of P-AMY increase did not significantly differ between HIV+ and HIV - populations (14.2% vs 15.2%, P = .91) or between HIV+Tx+ and HIV+Tx - (14.7% vs 8.9%, P = .11). In almost half (48.3% of HIV+ and 42.9% of HIV -) of hyperamylasemic patients, lipase was normal, indicating a non pancreatic origin of their P-AMY increase. Markedly elevated P-AMY (>3 times the URL) was found in six HIV+ patients and in one HIV - patient: two had macroamylasemia, one acute pancreatitis, three (including the HIV - patient) chronic pancreatitis, and one chronic hyperamylasemia of undefined origin.Conclusions: In our study, both HIV+ and HIV+Tx+ do not show an increased frequency of P-AMY elevation. Frank pancreatic disease is rare in this clinical setting.