Why the MDRD equation should not be used in patients with normal renal function (and normal creatinine values)?
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Biomedical subjects
Publications and source records attributed to J P Chapelle.
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Glomerular filtration rate (GFR) is the most frequently used parameter to evaluate the renal function. GFR may be estimated with serum creatinine, creatinine clearance based on 24 hours urine collection or Cockcroft formula. All these methods have bias. Other approaches have thus been proposed. The limitations and advantages of isotopic methods and recent mathematical approaches (MDRD formula) are reviewed.
Glomerular filtration rate (GFR) is the best indicator of renal function. GFR is usually estimated by serum creatinine or the creatinine clearance calculated on urine collected over 24 hours or with the Cockcroft formula. These methods are however limited. Serum creatinine has a very poor sensitivity and urine collection is difficult. Cystatin C is a protease inhibitor produced in a constant manner by nucleated cells. This molecule is freely filtrated by the glomerule and quite completely catabolized in the proximal tubules. Its plasmatic concentration might thus be used to estimate GFR. Presently available data allow to conclude that plasmatic cystatin C is at least as good as serum creatinine to estimate GFR. It is less sensible to changes in body mass. Its determination appears more sensitive to detect early mild changes in GFR. Reference values are presently available for the different methods of determination. Cystatin C plasma level determination is more expensive than routine creatinine plasma determination. In the absence of very significant advantages, this might explain its limited use in daily clinical practice.
Like other medical investigations, laboratory tests are not free from uncertainties or errors which limit their clinical value. Usually two levels of variations are identified: the analytical and biological levels. This paper presents some sources of variations and provides different examples. The concept of reference value is also discussed, as well as the clinician's viewpoint regarding the interpretation of laboratory results.
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We evaluated the AxSYM immunoassay for the quantification of cardiac troponin I (cTnI). Total assay imprecision, expressed as coefficient of variation, ranged between 5.6% and 8.3% for commercial control serum samples and between 4.2% and 13.9% for pooled patient samples. Linearity was verified up to 42 micrograms/L. Triglycerides (up to 1,000 mg/dL) did not interfere with the assay, but minor hemolysis and clinically relevant hyperbilirubinemia caused a negative bias. In 186 patient samples, AxSYM cTnI levels correlated significantly with data obtained with the Stratus II cTnI fluorometric enzyme immunoassay but were 3 to 4 times higher on AxSYM than on Stratus II. In 111 healthy blood donors, the reference range for cTnI levels on AxSYM was 0.0 to 0.4 microgram/L. After eccentric isokinetic exercise, healthy volunteers showed a rise in creatine kinase MB mass (AxSYM) but not in cTnI. On AxSYM and Stratus II, cTnI levels increased above the manufacturer's cutoff for acute myocardial infarction in all 17 patients followed up after onset of infarction-related chest pain but in only 1 of 91 control subjects. The AxSYM cTnI assay is a valid alternative for the detection of myocardial injury with diagnostic performance comparable to the established Stratus cTnI assay.
The troponin (Tn) complex consists of three subunits referred to as TnT, TnI and TnC. Myocardium contains TnT and TnI isoforms which are not present in skeletal muscles and which can be separated from the muscular isoforms by immunological techniques. Using commercially available immunoassays, clinical laboratories are able to determine cardiac TnT and TnI (cTnT and cTnI) quickly and reliably as classical cardiac markers. After acute myocardial infarction, cTnT and cTnI concentrations start to increase in serum in a rather similar way than CK-MB, but return to normal after longer periods of time (approximately one week). Because of their excellent cardiac specificity, Tn subunits appear ideally suited for the differential diagnosis of myocardial and muscular damage, for example in noncardiac surgery patients, in patients with muscular trauma or with chronic muscular diseases, or after intense physical exercise. cTnT and cTnI may also be used for detecting evidence of minor myocardial damage: therefore they have found new clinical applications, in particular risk stratification in patients with unstable angina. In spite of the possible reexpression of cTnT in human skeletal muscles, and of the lack of standardization of cTnI assays, Tn subunits are not far to meet the criteria of ideal markers for acute myocardial injury. Only an insufficient sensitivity in the first hours following the acute coronary syndroms requiries to maintain an early myocardial marker in the cardiac panel for routine laboratory testing.
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Homocystinuria is an uncommon genetic disease characterized by a marked increase of serum homocysteine (HCY), an intermediate of methionine metabolism. In patients with homocystinuria, hyperhomocysteinemia promotes the development of atherosclerotic lesions and is responsible for premature coronary artery disease. Recently, several studies have also demonstrated that moderate hyperhomocysteinemia--not necessarily linked to an inborn metabolic defect--may also be considered as an independant risk factor for cardiovascular disease. The main mechanisms of HCY atherogenic action are thought to be LDL oxydation, inhibition of vascular endothelium growth combined with stimulation of smooth muscular cells proliferation, and interference with the coagulation and fibrinolytic systems. Cofactors of key enzymes in HCY metabolism, folic acid, vitamin B12 and vitamin B6, may be given, alone or in combination, for the treatment of hyperhomocysteinemia. Homocysteinemia can be assessed by basal plasma HCY concentration and/or by HCY levels measured after a methionine loading test. Mainly measured till now in specialized laboratories using rather complex techniques (HPLC, GCMS, amino acid analyser ...), HCY determination is today spreading widely owing to the development of automated immunoassays.
Glycohaemoglobin, and particularly haemoglobin A1c(HbA1c), assays have been used for many years to retrospectively evaluate the glycaemic control of diabetic patients. Cut-off values have been established for deciding treatment modifications. The techniques used in the laboratories however exhibit varying quality, and all of them are not yet standardized. The consequence is an under-utilization of this test, especially in non-hospital practice. In this context, working groups of Société Française de Biologie Clinique (SFBC), Association de Langue Française pour l'Etude du Diabète et des Maladies Métaboliques (ALFEDIAM) and Société Française d'Endocrinologie (SFE) have met together, in order to analyze the national status, and to propose practical recommendations for implementing a standardization process on the basis of international experiences. It is recommended to exclusively express results as HbA1c percentage, using methods standardized and certified by comparison to reference methods such as those using Diabetes Control and Complications Trial (DCCT) values. Simultaneously, contacts have been established with manufacturers, and the realisation of periodic quality control surveys was encouraged.
A multicentre evaluation of the urine test strip analyser Super Aution-4220 was carried out in six laboratories. The analytical performance of the instrument with regard to imprecision, linearity, detection limit, drift, carry-over and method comparison was studied. Using the Aution stick 8 test strip the pH, glucose, protein, ketones, bilirubin, blood, urobilinogen and leukocyte esterase were analysed. Specific gravity measurements were performed by refractive index method. Within-run and between-run imprecision determined at three levels of analyte were good. No carry-over was observed. Obtained results were linear through all the described analytical range. No significant drift was detected. Method comparison with some quantitative methods was performed and showed a good correlation with most of the analytes. The study of interferences showed minor interferences by common therapeutic drugs with the measurement of some analytes. During the assessment period of about 6 months no breakdown occurred in any laboratory. The Super Aution urine analyser appeared to be a highly automated analyser of urinary test strips. The operation was simple and the maintenance required only a few minutes a day.
The troponin (Tn) complex consists of three protein subunits referred to as TnT, TnI and TnC. Myocardium contains TnI and TnT isoforms which are not present in skeletal muscles and which can be separated from the muscular isoforms by immunological techniques. Using commercially available immunoassays, clinical laboratories are able to determine cardiac TnT and TnI (cTnT and cTnI) as quickly and reliably as classical cardiac markers. After acute myocardial infarction, cTnT and cTnI concentrations start to increase in serum in a rather similar way than CK-MB, but return to normal after longer periods of time (approximately one week). Because of their excellent cardiac specificity, Tn subunits appear ideally suited for the differential diagnosis of myocardial and muscular damage, for example in non cardiac surgery patients, in patients with muscular trauma or with chronic muscular diseases, or after intensive physical exercise. cTnT and cTnI may also be used for detecting evidence of minor myocardial damage, and therefore may find new applications in the management of patients with unstable angina.
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Based on results from the Belgian External Quality Assessment (EQA) Scheme, we studied the main factors affecting the between-laboratory variation of C-reactive protein determination. Participants using homogeneous systems with several calibration points generally achieved better performance. Working temperatures influenced the results to a lesser extent. The present study stresses the importance for EQA organizers to collect more detailed information about CRP analytical methods used by the participants. It also suggests that manufacturers should be more involved in the management of quality, in particular by striving for standardization of the material (kit and calibrator) they produce for CRP assay.
A recently introduced turbidimetric immunoassay using shell/core particles for determination of myoglobin (Turbiquant Myoglobin, Behringwerke, Marburg, Germany) was evaluated on the Behring Turbitimer analyzer. Intra-assay CV varied between 1.0% and 3.2%; interassay CV was between 2.0% and 3.6%. Linear results were obtained between 2.8 nmol/L and 36.7 nmol/L. Manual dilution in saline solution allowed measurement of myoglobin concentrations up to 25,400 nmol/L. In the tested range, no high-dose 'hook' effect was observed. Haemolysis interfered with the assay when haemoglobin concentrations exceeded 2 g/L. The occurrence of error codes due to the presence of triglycerides was shown to depend on particle size. Repeat assays on diluted samples were necessary at triglyceride concentrations of 2.3 mmol/L for post-prandial chylomicrons (200-1000 nm), and at 11-31 mmol/L for very-low density lipoprotein-triglycerides (80-200 nm). No significant interferences of haptoglobin, bilirubin, iodine containing contrast media, and rheumatoid factors were detected for the assay. Treatment of lipaemic samples with Lipoclean (Behringwerke) resulted in lower myoglobin values. Simultaneously drawn serum and heparin-plasma samples gave comparable myoglobin results. However, values obtained in EDTA- or citrate-treated plasma samples occasionally differed significantly from serum values. The upper reference limit for myoglobin concentration was 4.2 nmol/L. The turbidimetric assay correlated well with a radioimmunoassay (Byk-Sangtec; r = 0.892) and with the automated nephelometric assay (Behringwerke; r = 0.944). Values obtained by turbidimetry were comparable to those obtained by the latex agglutination test (Behringwerke).
A recently introduced automated nephelometric immunoassay involving shell/core particles for determination of myoglobin (Behringwerke) was evaluated with the BNA Nephelometer. Method precision was good: the intra-assay CV varied between 1.5% and 6.1%; with daily calibration, the interassay CV ranged between 1.5% and 7.5%. For usual sample dilutions, the assay response varied linearly with myoglobin concentrations up to 23.1 nmol/L. After automatic dilution by the instrument, concentrations up to 2310 nmol/L could be measured without high-dose "hook" effect. Further manual dilution allowed measurement of myoglobin concentrations up to 26,000 nmol/L. Calibration was stable for at least seven days. We detected no significant interferences from hemoglobin, haptoglobin, bilirubin, iodine-containing contrast media, and rheumatoid factors. Treating lipemic samples with Lipoclean (Behringwerke) decreased test results. Simultaneously drawn serum and plasma samples from the same subject showed no consistent differences in myoglobin concentrations. The mean reference myoglobin concentration was 1.380 (SD 0.82) nmol/L for men and 0.878 (SD 0.45) nmol/L for women. In patients with renal insufficiency, serum creatinine values were moderately related to serum myoglobin values (r = 0.465). Although a commercial radioimmunoassay (Byk-Sangtec) and the nephelometric assay intercorrelated well (r = 0.929), values obtained by nephelometry were significantly lower (P less than 0.05). By both assays, results for heart and skeletal muscle tissue extracts showed no correlation, a finding that suggests the existence of multiple forms of myoglobin in human tissues. We conclude that immunonephelometry is a rapid, practical, and reliable method for measuring myoglobin in serum.
We evaluated the analytical and clinical performances of a new radial partition immunoassay for measuring the mass concentration of creatine kinase (CK)-MB in serum. All pipetting, washes, incubations and data reduction were performed in 8 min by the Stratus (Dade) fluorometric analyzer. Within-assay and between-assay CVs were respectively 5.5% and 8.4% at 21 micrograms/L, and 4.2% and 3.4% at 48 micrograms/L. Assaying serial dilutions of serum samples with high CK-MB concentrations demonstrated excellent linearity. Results of the Stratus technique correlated well (n = 115, r = 0.98) with those of the Tandem-E CKMB II assay. There was no interference from hemolysis, bilirubin, rheumatoid factor, or added CK-MM (up to 3500 U/L); consequently, CK-MB can be determined in undiluted serum, even in the presence of high total CK activity. The mean CK-MB concentration in 105 blood donors was 1.9 (SD 1.3) micrograms/L. For seven myocardial infarction patients who received prompt fibrinolytic therapy, the mean CK-MB concentration was 4.5 (SD 1.8) micrograms/L at admission, and maximum concentrations, 119 (SD 94) micrograms/L, were recorded 16 h later. CK-MB returned to concentrations less than 10 micrograms/L within 72 h.
To define the independent variables predictive of early versus late mortality after acute myocardial infarction (AMI), 420 consecutive patients were studied and divided into 3 groups: the 45 patients who died within the initial 3 months (group 1), the 45 patients who died greater than 3 months and less than or equal to 3 years after AMI (group 2) and the 330 greater than 3-year survivors (group 3). The stepwise logistic discrimination method was applied to clinical and laboratory variables recorded during hospitalization to distinguish among the 3 groups. Six independent variables were found to be predictive of early mortality: left ventricular function score (chi-square 26.2; p less than 0.00001), ventricular fibrillation (chi-square 9.3; p = 0.002), bundle branch block (chi-square 9.0; p = 0.003), history of previous AMI (chi-square 8.7; p = 0.003), age (chi-square 5.8; p = 0.02) and atrioventricular block (chi-square 3.8; p = 0.05). Three independent variables were found predictive of late mortality: age (chi-square 13.8; p = 0.0002), anterior location of the AMI (chi-square 4.0; p = 0.04) and a low peak creatine kinase-MB level (chi-square 3.8; p = 0.05). Only 2 variables were able to distinguish between early and late nonsurvivors: peak creatine kinase-MB level (chi-square 8.7; p = 0.003) and ventricular fibrillation (chi-square 4.6; p = 0.03). Thus, the sets of independent risk factors for early and late mortality after AMI are substantially different--suggesting that differing mechanisms are responsible for outcome.