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Detection of macroenzymes: establishing upper reference limits for eight enzymes after polyethylene glycol
Lieselot Dedeene1, Marie Stockman1, Sophie Steels1
1Clinical Department of Laboratory Medicine, University Hospitals Leuven, Leuven, Belgium.
Introduction:
The 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.
Materials And Methods:
Per 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°C and centrifuged. Supernatant enzyme activity was measured on Cobas c702 and the %PPA was calculated.
Results:
The 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 < 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%.
Conclusion:
In 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.
Insights
This study establishes upper reference limits for polyethylene glycol (PEG) precipitable activity (%PPA) in eight enzymes to aid in macroenzyme detection. Laboratory-specific %PPA limits are crucial for accurate interpretation and reducing unnecessary investigations.
Area of Science:
- Clinical Biochemistry
- Diagnostic Laboratory Medicine
Background:
- Macroenzymes in blood can lead to diagnostic confusion and unnecessary medical investigations.
- Polyethylene glycol (PEG) precipitation is a common method for detecting macroenzymes.
- A lack of consensus exists regarding upper reference limits for PEG-precipitable activity (%PPA) of monomeric enzymes.
Purpose of the Study:
- To establish laboratory-specific upper reference limits (URLs) for %PPA for eight enzymes using PEG precipitation.
- To determine the imprecision of the PEG precipitation method for these enzymes.
- To clinically verify the established URLs using samples with known macroenzymes.
Main Methods:
- Collected blood samples from adult patients with normal or elevated enzyme activities.
- Applied polyethylene glycol (PEG) 6000 precipitation to samples.
- Measured supernatant enzyme activity and calculated %PPA.
- Determined within-laboratory imprecision and clinically verified URLs with electrophoresis-identified macroenzyme samples.
Main Results:
- Established URLs for eight enzymes, including amylase (26% PPA), alkaline phosphatase (29% PPA), and creatine kinase (24% PPA).
- Demonstrated within-laboratory imprecision of < 15% for the PEG precipitation method.
- Clinical verification showed samples with macroCK and macroALP had %PPA values of 69%, 43%, and 52%, respectively.
Conclusions:
- Upper reference limits for monomeric enzyme %PPA after PEG precipitation were successfully established for eight enzymes.
- The established URLs are suitable for clinical use but require laboratory-specific validation.
- Highlight the importance of using laboratory-specific %PPA reference limits for accurate macroenzyme detection and interpretation.
