Journal of clinical chemistry and clinical biochemistry. Zeitschrift fur klinische Chemie und klinische Biochemie最新文献

This review discusses the interference in clinical chemical analysis caused by autoantibodies and by antibodies to foreign antigens. The nature of these disturbances, their occurrence, prevalence, and detection, the different ways in which they are manifested, the clinical consequences of the failure to recognize such interference, and finally methods for avoiding these disturbances are discussed. Interference by cold agglutinins in the automatic determination of the erythrocyte count, interference by cryoglobulins in the determination of the leukocyte count, and EDTA-induced thrombocyte agglutination are well documented as sources of error in the analysis of haematological parameters. Enzyme determinations may be affected by the occurrence of macro-complexes of the measured enzyme with immunoglobulins. This type of interference, for example, may occur in the determination of amylase and creatine kinase. Immunoassays for the determination of hormones or tumour markers are sensitive to autoantibodies and heterophilic antibodies. The former are particularly important in the determination of thyroid hormones, where the interference is method-dependent. In several immunoassays, interference by heterophilic antibodies can be abolished by the addition of non-immune serum. Finally, rheumatoid factors, antibodies administered for therapeutic purposes, and monoclonal gammopathies are possible sources of interference in the determination of various analytes.

A method for the separation and quantification of a complex ganglioside mixture from a clinically available amount (5 ml) of human cerebrospinal fluid (CSF) is described. After reduction of the CSF volume by ultrafiltration, gangliosides are extracted with methanol/chloroform, then separated and quantified by high performance thin layer chromatography (HPTLC) and direct densitometry. For purification of crude ganglioside extract, the method of choice was microdialysis against water. Recovery for the present method including all methodological steps was 78%. No delective loss of gangliosides was demonstrated. The CSF ganglioside pattern from 'normal' CSF samples resembles that of brain gangliosides, particularly cerebellum gangliosides. Based on chromatographic comparison with standards, the percentages of lipid-bound NeuAc-positive fractions were: GM1 = II3NeuAc-GgOse4Cer (3%), GD3 = II3NeuAc2-Lac-Cer (3%), GD1a = IV3NeuAc,II3NeuAc-GgOse4Cer (15%), X1 (3%), GD1b = II3(NeuAc)2-GgOse4Cer (16%), X2 (3%), GT1b = IV3NeuAc,II3NeuAc2-GgOse4-Cer (41%), and GQ1b = IV3NeuAc2-,II3NeuAc2-GgOse4-Cer (16%). The total ganglioside content varied between 616-944 micrograms/l. Within-run and between-run assay precision (relative standard deviation) for 'normal' pooled CSF ranged from 0.04 to 0.12 for the predominant CSF ganglioside fractions (GD1a, GD1b, GT1b, GQ1b), and from 0.13 to 0.23 for the less pronounced fractions (GM1, GD3).

The reliability of the determination of the most common substrates with the Ektachem 700 was evaluated. Accuracy control was performed in various ways including the comparison of results with reference method values. The influence of protein concentration was investigated systematically using sera containing varying amounts of protein obtained by ultracentrifugation. Bilirubin. Deviation from the reference method values was within limits of the guidelines (Dt. Arztebl. 85 (1988) B517-B532). The influence of protein concentration was negligible. Cholesterol. The results agreed well with the reference method values. The method was not influenced by different protein concentrations. Creatinine. The results were in good agreement with reference method values, when the method was calibrated with the primary assigned values of the calibrators. At high protein concentrations, the results were higher than those of the comparative method. Glucose. The mean deviation from the reference method values was 1.0%. There was a clear positive bias at high protein concentrations. Protein. The results from the Ektachem deviated from the reference method values by -6.8%. Total glycerol (triacylglycerols). In the analysis of control sera, the Ektachem values differed greatly (+32.1%) from the reference method values. This difference was not found in a comparative study with native sera. At high protein concentrations the Ektachem results were higher than those of the comparative method. Urea. The results were lower than the method-dependent assigned values (-16.6%). This deviation was not observed in a comparative study with native sera. Interference by protein was not observed. Uric acid. In accuracy control with reference method values, a small bias was observed (+3.3%), which increased at high protein concentrations.

An elevated erythrocyte sedimentation rate is generally regarded as an unspecific and mostly pathological indicator of inflammation or tumour. However, we have determined the concentrations of plasma/serum proteins that influence the erythrocyte sedimentation rate in numerous samples from several groups of patients with different diseases, including 2 forms of cancer. Equations have been developed by which the 1 h value of erythrocyte sedimentation rate can be expressed as the sum of disease-specific coefficients for each protein multiplied by the measured concentrations of the respective proteins. These equations are shown to be disease-specific with 64-93% probability. Such equations may thus form the basis for differential diagnosis.

Substance concentrations of plasma cholinesterase (EC 3.1.1.8) were measured in 94 healthy individuals without occupational exposure to known inhibitors (six samples from each individual). Immunoreactive cholinesterase substance concentrations showed an inter-individual variation corresponding to CVtotal = 22% (mean: 5.01 mg/l, SD: 1.11 mg/l). Intra-individual variations of immunoreactive cholinesterase substance concentration were correlated (r = 0.36) to intra-individual variation of albumin. Estimated by a repeated-measures analysis of variance, the observed intra-individual variation of cholinesterase substance concentration corresponded to CV = 8.8% (SD: 0.44 mg/l), which together with a CVerror = 6% (within and between runs), implies a biological intra-individual variation of cholinesterase substance concentration corresponding to CVintra = 6.4%. Specific catalytic activity (kU/mg immunoreactive cholinesterase) was influenced by the ChE-1 phenotype (phenotype U: 1.58 kU/mg, phenotype UA: 1.22 kU/mg), but not by body weight, height, age, and sex. Observed intra-individual variation of specific catalytic activity corresponded to 6.4% (SD: 0.10 kU/mg), which together with an estimated CVerror = 6.2% implies the biological intra-individual variations of specific catalytic cholinesterase activity to be insignificant. The insignificant CVintra makes specific catalytic cholinesterase activity a rational quantity for evaluation of unexpected fluctuations of cholinesterase activity concentrations.

The reliability of electrolyte determinations with the Ektachem 700 was evaluated by various means including the use of reference method values. The influence of protein concentration, which may alter the viscosity and hence the speed of diffusion, was systematically investigated by using samples of varying protein concentration obtained by ultracentrifugation. Calcium. The mean bias between Ektachem results and reference method values of 9 control sera was -7.3%. In a comparative study with native sera a negative bias was not obtained. The accuracy was independent of the protein concentration. Chloride. The mean bias was -0.1% compared with reference method values. As might be expected, the difference between chloride values with Ektachem--using a "direct" ion-selective electrode--and determinations by coulometry increased with increasing protein concentrations. Magnesium. A negative bias (-5.0%) was obtained in accuracy control with reference method values, and to a smaller extent in a comparative study with native sera. An influence of the protein concentration on the magnesium determination was not observed. Phosphate. Accuracy control by method-dependent assigned values and a comparative study with native sera showed a positive bias. Ektachem results depend on the protein concentration. At 120 g/l protein the bias was +13.6%. Potassium. The mean bias, with respect to reference method values, was -1.2%. At high concentrations of proteins of "normal" composition, Ektachem results agree with measurements by flame atomic emission spectrometry; in paraproteinaemic sera the values are higher. Sodium. The mean inaccuracy was 3.3%, compared with reference method values. The dependence on the amount and composition of the total protein was similar to that found for potassium analysis.

We present a radiometric assay for the determination of urinary angiotensin-converting enzyme activity, using benzoyl-[1-14C]glycyl-L-histidyl-L-leucine as the substrate. An optimal pH of 8.3, an optimal chloride concentration of 0.375 mol/l and complete inhibition by EDTA-Na2, captopril and enalaprilat confirm the specificity of the assay. Comparison of dialysis and ultrafiltration for concentration of urine showed the existence of angiotensin-converting enzyme inhibitors in human urine. Dialysis against water was the more effective method for avoiding enzyme inhibition. After dialysis of urine, the assay was linear with time and with enzyme concentration; it was highly sensitive (60 mU/l) and showed good reproducibility. Under our technical conditions, we found angiotensin-converting enzyme activity in urine samples with quantitatively abnormal protein contents, but not in normal urine. Urinary angiotensin-converting enzyme did not correlate with proteinuria nor with water-salt parameters or creatinine. We confirm the kidney tubular epithelial origin of the enzyme, and propose the use of our assay to study urinary angiotensin-converting enzyme as a marker of renal tubular damage.

alpha 1-Microglobulin was purified from urine to a purity of 97.7% in a yield of 25.8%, and was used to produce antibodies in sheep. These antibodies, purified by affinity chromatography, were used to develop a rapid one-step and a two-step immunoenzymometric assay (IEMA). The equilibrium in the reaction between solid phase-adsorbed antibodies and antigen and between the antigen and enzyme-labelled antibodies was attained within 30 and 100 min, respectively. The one-step IEMA permits a good differentiation of low alpha 1-microglobulin concentrations after 30 min reaction time. Its detection limit is 0.35 micrograms/l, and its measurement range is between 0.5 and 100 micrograms/l. The IEMA correlates well with radial immunodiffusion (r = 0.973). The mean alpha 1-microglobulin serum concentration in women is insignificantly lower (33.2 mg/l) than in men (36.1 mg/l). In both sexes the alpha 1-microglobulin concentration increases with age. HIV-infected symptomless men have a significantly lower (15.9 mg/l) alpha 1-microglobulin concentration in serum than normal persons, whereas in AIDS patients it is significantly higher (45.5 mg/l).