{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T12:19:13Z","timestamp":1790943553354,"version":"4.1.0"},"reference-count":24,"publisher":"Croatian Society for Medical Biochemistry and Laboratory Medicine","issue":"1","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biochem. med. (Online)"],"published-print":{"date-parts":[[2023,2,15]]},"abstract":"<jats:p>IntroductionThe presence of macroenzymes in blood can cause diagnostic confusion. Therefore, confirming the presence of macroenzymes is important to reduce unnecessary (non-)invasive investigations. Polyethylene glycol (PEG) precipitation is a simple and fast first-line method for the detection of macroenzymes. However, there is no consensus on the upper reference limit for the PEG-precipitable activity (%PPA) of monomeric enzymes. The aim of this study was to verify a PEG precipitation protocol for the detection of macroenzymes in our laboratory by establishing upper reference limits (URLs) and determining imprecision for eight enzymes after PEG precipitation. In addition, we aimed to clinically verify the URLs using samples containing macroenzymes as identified by electrophoresis.\nMaterials and methodsPer enzyme, at least 40 leftover blood samples from adult patients with either normal or increased enzyme activities were diluted 1:1 with 25% PEG 6000 and 1:1 with 0.9% NaCl. Mixtures were incubated for 10 min at 37\u00b0C and centrifuged. Supernatant enzyme activity was measured on Cobas c702 and the %PPA was calculated.\nResultsThe following URLs were obtained: 26% PPA for amylase, 29% PPA for alkaline phosphatase (ALP), 61% PPA for alanine aminotransferase, 48% PPA for aspartate aminotransferase, 24% PPA for creatine kinase (CK), 55% PPA for gamma-glutamyltransferase, 65% PPA for lactate dehydrogenase, and 56% PPA for lipase. The within-lab imprecision was &lt; 15%. Regarding the clinical verification, the two historical samples with proven macroCK showed a %PPA of 69% and 43%, respectively, and a sample with proven macroALP had a %PPA of 52%.\nConclusionIn this study, URLs for monomeric enzyme activities after PEG precipitation for eight different enzymes were established. The URLs are suitable for clinical use, but are only partially in line with other studies. Therefore, our data highlight the importance of establishing laboratory-specific upper reference limits for %PPA to allow a correct interpretation.<\/jats:p>","DOI":"10.11613\/bm.2023.010705","type":"journal-article","created":{"date-parts":[[2022,12,19]],"date-time":"2022-12-19T12:47:46Z","timestamp":1671454066000},"source":"Crossref","is-referenced-by-count":5,"title":["Detection of macroenzymes"],"prefix":"10.11613","volume":"33","author":[{"given":"Pieter","family":"Vermeersch","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Glynis","family":"Frans","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lieselot","family":"Dedeene","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie","family":"Stockman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sophie","family":"Steels","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"4885","published-online":{"date-parts":[[2022,12,15]]},"reference":[{"key":"key20221219074722_r1","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1258\/000456303322326425","article-title":"Macroenzyme detection by polyethylene glycol precipitation.","volume":"40","author":"Davidson","year":"2003","journal-title":"Ann Clin Biochem"},{"key":"key20221219074722_r2","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1016\/S0016-5085(82)80331-4","article-title":"A Rapid and Simple Assay to Determine if Macroamylase Is the Cause of Hyperamylasemia.","volume":"83","author":"Levitt","year":"1982","journal-title":"Gastroenterology"},{"key":"key20221219074722_r3","doi-asserted-by":"crossref","first-page":"2052","DOI":"10.1016\/j.cca.2011.01.023","article-title":"Detection of macro-creatine kinase and macroamylase by polyethylene glycol precipitation and ultrafiltration methods.","volume":"412","author":"Wyness","year":"2011","journal-title":"Clin Chim Acta"},{"key":"key20221219074722_r4","first-page":"65","article-title":"Macro enzymes: prevalence, composition, detection and clinical relevance.","volume":"28","author":"Sanders","year":"1990","journal-title":"Clin Chem Lab Med"},{"key":"key20221219074722_r5","doi-asserted-by":"crossref","first-page":"1161","DOI":"10.1016\/j.cca.2010.11.005","article-title":"Reference intervals for six enzymes after polyethylene glycol precipitation and ultrafiltration.","volume":"412","author":"Wyness","year":"2011","journal-title":"Clin Chim Acta"},{"key":"key20221219074722_r6","first-page":"392","article-title":"Macroamylasemia and other immunoglobulin-complexed enzyme disorders.","volume":"133","author":"Klonoff","year":"1980","journal-title":"West J Med"},{"key":"key20221219074722_r7","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1309\/AJCPWA9QHR9WHPNA","article-title":"Multiple macroenzymes in a patient with AIDS: Diagnosis using ultrafiltration.","volume":"142","author":"Wyness","year":"2014","journal-title":"Am J Clin Pathol"},{"key":"key20221219074722_r8","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1258\/acb.2008.007233","article-title":"False-positive polyethylene glycol precipitation tests for macroprolactin due to increased serum globulins.","volume":"45","author":"Ram","year":"2008","journal-title":"Ann Clin Biochem"},{"key":"key20221219074722_r9","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1515\/cclm-2020-0804","article-title":"The new IVD Regulation 2017\/746: A case study at a large university hospital laboratory in Belgium demonstrates the need for clarification on the degrees of freedom laboratories have to use lab-developed tests to improve patient care.","volume":"59","author":"Vermeersch","year":"2020","journal-title":"Clin Chem Lab Med"},{"key":"key20221219074722_r10","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1515\/CCLM.1998.034","article-title":"Approved recommendation on IFCC methods for the measurement of catalytic concentration of enzymes. 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