Hemolysis interference studies: freeze method should be used in the preparation of hemolyzed samples

Gidske, Gro; Sølvik, Una Ørvim; Sandberg, Sverre; Kristensen, Gunn B.B.

doi:10.1515/cclm-2018-0193

Cited by 12 publications

(14 citation statements)

References 7 publications

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“…For hemolysis interference, the results of quantitative analysis are largely consistent with previous reports, including changes in LDH, potassium, CK, CK‐MB, and some other analytes 12,18,19 . However, several previous studies used freeze‐and‐thaw or mechanical trauma without assessing the achievement of cell lysis 12,16,20,21 . In these studies, the extent of hemolysis was solely determined from the hemolytic index measured by the machine.…”

Section: Discussionsupporting

confidence: 85%

“…12,18,19 However, several previous studies used freeze-and-thaw or mechanical trauma without assessing the achievement of cell lysis. 12,16,20,21 In these studies, the extent of hemolysis was solely determined from the hemolytic index measured by the machine. However, it is impractical for clinical staff to determine whether a clearly hemolyzed blood sample is acceptable because the hemolytic index cannot be Therefore, the reference chart is even more significant at these laboratories as they can simply compare the reddish colors of the sample and the standards listed on the chart for estimation.…”

Section: Discussionmentioning

confidence: 99%

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A Reference chart for clinical biochemical tests of hemolyzed serum samples

Zhu

Wang

et al. 2020

Clinical Laboratory Analysis

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Background Although the effect of hemolysis has been extensively evaluated on clinical biochemical tests, a practical guidance for laboratory staff to rapidly determine whether a hemolyzed blood sample is acceptable and how to interpret the results is lacking. Here, we introduce a chart as a convenient reference for dealing with such samples. Methods Serum samples with 0.1%, 0.3%, 1%, 3%, and 10% hemolysis were prepared from sonicated endogenous red blood cells and received 35 wet and 22 dry clinical biochemical tests, respectively. The contributing part in the biochemical test result at each hemolysis condition was derived by subtracting the original test result of this sample with no hemolysis. The net results were used for analyses and preparation of the reference chart. Results The reference chart displayed the analytically calculated hemolysis interference and related statistical analyses. The chart also provided the color appearance of serum samples at each hemolysis condition for clinical staffs to determine whether a hemolyzed sample could be accepted. Conclusion In clinical laboratories, preparation of such a reference chart is extremely useful in dealing with hemolyzed blood samples for clinical biochemical tests.

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Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 99%

A Reference chart for clinical biochemical tests of hemolyzed serum samples

Zhu

Wang

et al. 2020

Clinical Laboratory Analysis

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“…The most commonly used methods for preparation of hemolysate for interference studies is osmotic shock, freezing/thawing, and shearing with multiple needle aspirations . Recently, a comparative study of these three approaches was performed by Gidske et al They concluded that the freeze/thaw method is to prefer since it includes lysis of leukocytes, platelets, and erythrocytes and is less laborious than the needle aspiration technique. Another approach is to ask for a second replacement sample for a hemolyzed patient sample .…”

Section: Introductionmentioning

confidence: 99%

Hemolysis interference in 10 coagulation assays on an instrument with viscosity‐based, chromogenic, and turbidimetric clot detection

Hedeland

Gustafsson

Touza

et al. 2020

Int J Lab Hematology

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Introduction: Hemolysate in plasma samples from patients may cause misleading results in coagulation assays. Even though modern coagulation instruments often are equipped with modules that can detect hemolysis, icterus, and lipemia (HIL), studies that report the influence of these interferences are still limited. The present paper focuses on the influence of hemolysis on 10 coagulation assays.Methods: Artificial hemolysis was created by freezing/thawing, and the hemolysates generated were added to pools of patient plasma. Pathological and normal levels were pooled separately. These spiked samples were analyzed on a STA R Max 2 instrument. The coagulation assays evaluated utilize clot, chromogenic, or immunoturbidimetric detection. Results:Four of the evaluated assays were not influenced by hemolysis: fibrinogen, von Willebrand factor antigen, activated partial thromboplastin time, and factor VIII.Interestingly, normal and slightly elevated prothrombin time (INR < 2.0) was insensitive to hemolysis, whereas samples with a high INR (≥2.0) exhibited falsely high readings. The assays for antithrombin and fibrin D-dimer displayed an intermediate sensitivity to hemolysis. The most sensitive assay turned out to be anti-Xa, followed by protein C and protein S. For the anti-Xa assay, the results are decreased by 10% already at 0.5 g/L hemoglobin. Conclusion:The present study shows that hemolysis affects several of commonly used coagulation assays. Since the sensitivity for hemolysis is dependent on the brand of the assay as well as the instrument and principle of measurement, it is necessary to evaluate the influence of each specific combination. K E Y W O R D Scoagulation, freezing, hemoglobin interference, hemolysis, pre-analytical error, serum indices

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“…The tubes were thawed for 1 h at 21 °C and centrifuged for 10 min at 1300 g at 21 °C. The top layers from the eight tubes were mixed and used as hemolysate [11,12]. The Hb concentration in the hemolysate and the four identical samples distributed to the participants was measured using a point-of-care instrument HemoCue Hb 201 + (HemoCue, Ängelholm, Sweden).…”

Section: Preparation Of Samplesmentioning

confidence: 99%

Handling of hemolyzed serum samples in clinical chemistry laboratories: the Nordic hemolysis project

Gidske¹,

Aakre

Rustad³

et al. 2019

Clinical Chemistry and Laboratory Medicine (CCLM)

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Background Some clinical chemistry measurement methods are vulnerable to interference if hemolyzed serum samples are used. The aims of this study were: (1) to obtain updated information about how hemolysis affects clinical chemistry test results on different instrument platforms used in Nordic laboratories, and (2) to obtain data on how test results from hemolyzed samples are reported in Nordic laboratories. Methods Four identical samples containing different degrees of hemolysis were prepared and distributed to 145 laboratories in the Nordic countries. The laboratories were asked to measure the concentration of cell-free hemoglobin (Hb), together with 15 clinical chemistry analytes. In addition, the laboratories completed a questionnaire about how hemolyzed samples are handled and reported. Results Automated detection of hemolysis in all routine patient samples was used by 63% of laboratories, and 88% had written procedures on how to handle hemolyzed samples. The different instrument platforms measured comparable mean Hb concentrations in the four samples. For most analytes, hemolysis caused a homogenous degree of interference regardless of the instrument platform used, except for alkaline phosphatase (ALP), bilirubin (total) and creatine kinase (CK). The recommended cut-off points for rejection of a result varied substantially between the manufacturers. The laboratories differed in how they reported test results, even when they used the same type of instrument. Conclusions Most of the analytes were homogeneously affected by hemolysis, regardless of the instrument used. There is large variation, however, between the laboratories on how they report test results from hemolyzed samples, even when they use the same type of instrument.

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Hemolysis interference studies: freeze method should be used in the preparation of hemolyzed samples

Cited by 12 publications

References 7 publications

A Reference chart for clinical biochemical tests of hemolyzed serum samples

A Reference chart for clinical biochemical tests of hemolyzed serum samples

Hemolysis interference in 10 coagulation assays on an instrument with viscosity‐based, chromogenic, and turbidimetric clot detection

Handling of hemolyzed serum samples in clinical chemistry laboratories: the Nordic hemolysis project

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