Interference by antiruthenium antibodies in the Roche thyroid-stimulating hormone assay.

There are many causes of interference in immunoassays causing erratic patient results. A method-specific interference due to antiruthenium antibodies in Roche free thyroxine (fT4) and free triiodothyronine (fT3) assays has been described previously. As a result, a new generation fT4 assay has been introduced by Roche. We describe six cases of interference due to antiruthenium antibodies, where in four cases interference in the Roche thyroid-stimulating hormone (TSH) assay was found as well. This raised the question as to whether other assays on this platform would also give incorrect results in patients with antiruthenium antibodies. Interference due to antiruthenium antibodies was suspected because of discrepancies between clinical presentation and/or TSH, fT4 and fT3 results. Samples of these six patients were re-analysed in Roche Diagnostics Laboratory, where it was demonstrated that the found discrepancies were indeed caused by interfering antiruthenium antibodies. Subsequently, these patients were asked to donate some blood once more for further evaluation, and three subjects agreed to participate. Their plasma was used to assay 18 analytes on Modular E and on a ruthenium-independent platform. The results were compared taking into account the known differences between distinct methods. As expected, significant interference was found in TSH. Also, in the new generation fT4 assay, ruthenium-induced interference was still present. However, the other assays, both competitive and immunometric, did not show clear interference. We therefore conclude that although antiruthenium antibodies theoretically can interfere in all assays on the Modular E platform, this kind of interference is found in the thyroid hormone assays, without marked interference in the other assays.

Patients' representations of their end-stage renal disease: relation with mortality.

BACKGROUND: Self-regulation theory explains how patients' illness perceptions influence self-management behaviour (e.g. via adherence to treatment). Following these assumptions, we explored whether illness perceptions of ESRD-patients are related to mortality rates. METHODS: Illness perceptions of 182 patients participating in the NECOSAD-2 study in the period between December 2004 and June 2005 were assessed. Cox proportional hazard models were used to estimate whether subsequent all-cause mortality could be attributed to illness perception dimensions. RESULTS: One-third of the participants had died at the end of the follow-up. Mortality rates were higher among patients who believed that their treatment was less effective in controlling their disease (perceived treatment control; RR = 0.71, P = 0.028). This effect remained stable after adjusting for sociodemographic and clinical variables (RR = 0.65, P = 0.015). CONCLUSIONS: If we consider risk factors for mortality, we tend to rely on clinical parameters rather than on patients' representations of their illness. Nevertheless, results from the current exploration may suggest that addressing patients' personal beliefs regarding the effectiveness of treatment can provide a powerful tool for predicting and perhaps even enhancing survival.

Unexpected prolonged extreme hypocalcaemia and an inadequate PTH response in a patient with metastatic breast carcinoma.

Although hypercalcaemia is often encountered during the course of malignant disease, hypocalcaemia appears to be rather rare. We describe a 37-year-old patient with metastatic carcinoma of the breast, who developed extreme hypocalcaemia (as low as 0.75 mmol calcium per litre) after chemotherapy. This is caused by a combination of hungry-bone syndrome and an insufficient parathyroid response. The latter may be the result of a direct toxic effect of chemotherapy on parathyroid hormone (PTH) synthesis possibly in combination with microscopic tumour infiltration in the parathyroid glands. Correction of the extreme hypocalcaemia over a period of 100 days by oral and intravenous calcium supplementation, corresponding to a total of 352 gram elemental calcium (1/3 of the total body calcium), resulted in gradual symptomatic relief. The possible mechanisms for these findings are discussed and the literature is briefly reviewed.

The influence of ketoacids on plasma creatinine assays in diabetic ketoacidosis.

OBJECTIVE: Analysis of the interference of ketoacids on various routine plasma creatinine assays during a clinical episode of diabetic ketoacidosis (DKA). DESIGN: Observational study. Blood samples were drawn before, during and after standard in-hospital treatment. Plasma creatinine was measured with two dissimilar enzymatic assays (creatininase PAP + and creatinine iminohydrolase Serapak), a kinetic alkaline picrate method (Jaffé) and a high-performance liquid chromatography (HPLC) procedure. Acetoacetate and beta-hydroxybutyrate were analysed by enzymatic methods. SETTING: Department of Medicine, University Hospital. SUBJECTS: Nine patients who experienced 10 episodes of DKA. MAIN OUTCOME MEASURES: Agreement of the routine plasma creatinine assays with HPLC and analysis of possible interferents. RESULTS: At presentation, the Jaffé assay gave falsely high values of plasma creatinine (median 99 micromol L(-1)), in contrast to the PAP+ (median 60.5 micromol L(-1)) and HPLC assays (median 67.5 micromol L(-1)). This positive error decreased during treatment. This was due to a decrease in acetoacetate, as the positive error by the Jaffé method correlated with the acetoacetate concentration (r = 0.79, P < 0.0001). In the multiple regression analysis, beta-hydroxybutyrate caused no additional interference by the Jaffé assay, confirmed by in vitro experiments. Analysis of agreement showed that the difference between PAP+ and HPLC creatinine was -4.6 +/- 3.0 micromol L(-1) (mean +/- SD), and 2.0 +/- 5.3 micromol L(-1) between Serapak and HPLC. This was statistically significant, but clinically negligible. CONCLUSION: Acetoacetate caused severe interference of the alkaline picrate (Jaffé) assay, which might influence therapeutic decisions at the start of diabetic ketoacidosis. Enzymatic assays lack this interference.

Independent prognostic value of C-reactive protein and troponin I in patients with unstable angina or non-Q-wave myocardial infarction.

OBJECTIVES: Elevated concentrations of C-reactive protein (CRP), a non-specific acute phase reactant, and troponin I (TnI), a cardiac-specific marker of myocardial damage, have been found to be associated with a higher risk for cardiac events in patients with an acute coronary syndrome. We evaluated CRP alone and in combination with TnI for predicting the incidence of major cardiac complications within 6 months in patients with unstable angina or non-Q-wave infarction (NQMI). METHODS: CRP and TnI was measured on admission in patients with unstable angina or NQMI, but results were kept blinded. Patients were treated according to a conservative management strategy, and the incidence of major cardiac events within 6 months was assessed. RESULTS: An abnormal CRP (> 5 mg/l) and an abnormal TnI (> 0.4 microgram/l) were more frequent in patients that suffered a major cardiac event (CRP: 93 vs. 35%, P < 0.0001; TnI: 73 vs. 26%, P < 0.001). The incidence of major cardiac events was higher in patients with an abnormal CRP than in patients with a normal CRP, both when TnI was abnormal (42 vs. 4.5%, P = 0.003) and when TnI was normal (11 vs. 0%, P = 0.014). Mean event-free survival was excellent in patients with both a normal CRP and TnI, whereas survival was poorest in patients with both an abnormal CRP and TnI (121 +/- 16 vs. 180 days, P < 0.0001). CONCLUSIONS: An abnormal CRP on admission in patients with unstable angina or NQMI is associated with increased incidence of major cardiac events within 6 months, both in patients with normal and abnormal TnI. CRP and TnI have independent and additive prognostic value in this patient group, and the combination may be useful for early risk stratification.

Clinical evaluation of the CARDIAC STATus, a rapid immunochromatographic assay for simultaneous detection of elevated concentrations of CK-MB and myoglobin in whole blood.

We studied the performance of the CARDIAC STATus, a new rapid, easy to perform qualitative whole blood bedside test for detection of elevated CK-MB and myoglobin in the emergency room. Blood samples from 182 consecutive patients with chest pain were drawn on admission and at five and seven hours after the onset of symptoms. The CARDIAC STATus tests were performed by coronary care unit nurses and, independently, by a trained laboratory technician. The results were compared with quantitative assays for CK-MB mass and myoglobin. At the end of the study, a second test series using a new lot number of cartridges was performed on the same blood samples because of possible elution buffer contamination. Nurses produced more false negative results than the technician (CK-MB 43 vs. 27 %, p=0.01, myoglobin 31 vs. 13%, p<0.0001), but the technician produced more false positive myoglobin results (9.3 vs. 5.5%, p=0.0001). In the second test series, the nurses produced significantly fewer false negative tests both for CK-MB (19%, p<0.0001) and myoglobin (13%, p=0.0002). The false negative rate for the technician was not different between the first and the second test series. The CARDIAC STATus yields a substantial number of false negative results both for CK-MB and myoglobin when compared to a quantitative assay, and therefore at present has limited value for ruling out an acute myocardial infarction.

Different hepatitis C virus (HCV) RNA load profiles following seroconversion among injecting drug users without correlation with HCV genotype and serum alanine aminotransferase levels.

Hepatitis C virus (HCV) infection often persists in association with chronic hepatitis. Different factors have been proposed to determine the clinical outcome of HCV infection. The aim of this study was to examine three different factors of HCV infection among injecting drug users. Nineteen untreated HCV seroconverters were tested longitudinally for the presence of HCV RNA by reverse transcriptase (RT) PCR, and results were quantified by the branched-DNA (bDNA) assay. HCV genotypes were determined with the first sample taken after HCV seroconversion. To assess the natural course of infection, serum alanine aminotransferase (ALT) levels were measured at three stages in every individual. The concordance between bDNA and RT-PCR was 98.9%. Three distinct patterns were found, according to the HCV RNA load after seroconversion during a mean follow-up period of 5 years (range, 1 to 8 years). HCV genotype 1a was predominant (52.6%). There was a significant increase in serum ALT levels (mean 55.5 U/liter) in the early phase of HCV infection, compared with basal serum ALT levels before HCV seroconversion and at the end of the follow-up period. Three distinct HCV RNA load profiles were found, without apparent relationship to genotype and serum ALT levels in the first 5 years of HCV infection.

[2nd revision consensus on blood transfusion. Central Guidance Organization for Peer Review]. / Tweede herziening consensus bloedtransfusie. Centraal Begeleidingsorgaan voor de Intercollegiale Toetsing.

Since 1982 several consensus conferences regarding the transfusion practice in hospitals have been organized in the Netherlands. Repeated updating of the consensus text such as described in this article is required to keep abreast of new developments and changes in clinical practice. Guidelines concerning compatibility testing (the result of compatibility testing is valid for three days at most), the organization of responsibilities and the clinical use of red cell concentrates, including a protocol for transfusion in patients with massive bleeding, were changed. No consensus was reached concerning the need to use only red cells that are antigen c, E and K compatible in women younger than 45 years of age and in patients with abnormal erythropoiesis. Although the need of systematic reviews to support guidelines was recognized, literature reviews revealed that only few studies were designed well enough to allow definitive conclusions regarding the benefits and risks of red cell transfusion.

Critical difference between serial measurements of CK-MB mass to detect myocardial damage.

To assess the critical difference in serial measurements of CK-MBmass and the ability of this critical difference to detect myocardial damage, we studied 110 patients in whom an acute myocardial infarction (AMI) had been ruled out. Blood samples were drawn at 3, 4, 5, 6, 7, 8, 12, 16, 20, and 24 h after onset of symptoms. With a critical difference of 72.6%, an increase of >2.0 microg/L between two CK-MBmass measurements was determined to be significant. Twenty-three of the non-AMI patients had an increase in CK-MBmass >2.0 microg/L, but five of these did not have an abnormal concentration of troponin T (i.e., not >0.1 microg/L). Also among the 110 non-AMI patients, 22 did have an abnormal troponin T value, 18 of whom (82%) also had CK-MBmass increased by >2.0 microg/L. In 20 of the 23 patients with an increase in CK-MBmass >2.0 microg/L, this increase was detected from the values for two samples collected at 5 and 12 h after onset of symptoms. In conclusion, using the critical difference for CK-MBmass defined as an increase >2.0 microg/L detected myocardial damage in patients without AMI.

Risk factors and anti-HBc reactivity among first time blood donors.

BACKGROUND AND OBJECTIVES: The usefulness of testing for antibody to hepatitis B core antigen (anti-HBc) as a surrogate marker for non-A, non-B hepatitis can no longer be clearly established in the face of anti-hepatitis C virus testing. Application of anti-HBc testing in blood donors for detection of hepatitis B in addition to hepatitis B surface antigen testing (HbsAg) is a matter of debate. MATERIALS AND METHODS: We examined the serology and risk analysis data in a group of first-time blood donors. In 1.48% of 16,081 donors, anti-HBc reactivity was found. We invited a study group of 112 donors for extensive interviewing about the risk of blood transmissible diseases, and for serological testing. A control group of 240 first-time donors was studied as well. RESULTS: In the study group, the age was older (p < 0.001), a history of liver disease was more frequent (p < 0.001), and the donor (p < 0.001) or the donor's partner (p < 0.05) had either stayed longer in an HBV-endemic area or had been born in one. Combining these with the serological results, we found that strong anti-HBc reactivity was related to hepatitis B risk factors in HBsAg-negative donors. CONCLUSIONS: Anti-HBc testing in HbsAg-negative first-time donors makes it possible to identify hepatitis B risk factors with a prevalence of 0.02%. Our findings also stress the importance of including the history of the donor's partner(s) in the risk analysis before blood donation.

Intensification of donor interviewing procedures: a feasibility study.

OBJECTIVE: To determine the feasibility and acceptability for the blood donor of an intensified blood donor interviewing procedure on high-risk factors for infectious diseases. To answer the question whether an intensified blood donor interviewing procedure would lead to an unacceptable loss of blood donors. DESIGN: Feasibility study. SETTING: Red Cross Bloodbank Rotterdam. DONORS: Study group of 240 first-time donors. INTERVENTIONS: Intensified donor interviewing techniques by direct questioning and workload assessment. RESULTS: Intensified interviewing was welcomed by 88-91% of first-time donors and rejected by 2-5%. On the question whether the intensified interviewing procedure should be the standard approach of the blood bank the answer was positive in 76-82% of first-time donors and negative in 11-14%. No blood donors indicated that this would be a reason to withdraw from blood donation. The workload for the blood bank physician increased by approximately 30%. CONCLUSION: The approach of intensified donor interviewing techniques in first-time donors is acceptable both to the donors and the blood bank workload.

Exclusion of acute myocardial infarction. The value of measuring creatine kinase slope.

For the exclusion (and diagnosis) of acute myocardial infarction, we studied timed sequential (slope) measurements of creatine kinase and creatine kinase-MB catalytic activity concentration, creatine kinase-MB mass concentration, troponin T and myoglobin, using data from 242 patients consecutively admitted for evaluation of suspected acute myocardial infarction in the 12 hours before admission. Three biochemical strategies based on measurements in two consecutive samples obtained within 12 hours after admission were evaluated. The highest sensitivities were encountered for a biochemical strategy based on the sole measurement of creatine kinase mass concentration (98%) or troponin T (96%) and a strategy based on measurements of creatine kinase activity concentrations, which includes creatine kinase slope calculation and measurement of creatine kinase mass concentration (95%). Both strategies were applied in subgroups of patients based on the electrocardiographic findings. In patients with a normal electrocardiogram, the sensitivity of the strategy using sole measurements of creatine kinase mass concentration was 100%, but this was true for the strategy based on creatine kinase slope measurements, which is the cheaper and therefore preferred procedure for excluding myocardial infarction. This approach, however, does not account for detecting minor myocardial cell damage in patients not yet fulfilling the criteria of the World Health Organization for diagnosing acute myocardial infarction.

Evaluation of the Hitachi 911 for routine urine analysis and for measurement of various special serum analytes.

The Boehringer Mannheim Hitachi 911 is a selective analyzer for 35 different methods including 3 ion-selective electrode (ISE) methods. We have evaluated this analyzer primarily to obtain objective information on its applicability for routine urine analyses in our laboratory. We also implemented appropriate assays for various special serum- and whole blood-tests, some for the first time on the Hitachi 911 and some with modified settings. Analytical evaluation involved NCCLS EP5-T2 (imprecision), NCCLS EP6-P (linearity), Krouwer 27 (multifactor) and Passing & Bablok (method comparison) evaluation protocols. With the exception of evidence of systematic erroneous sample predilution, overall results were favourable. Practicability of the Hitachi 911 was judged by simulating daily routine. During a period of two weeks, daily urine samples were rerun on the Hitachi 911, leading to a gain of about 50% in total processing time. It was concluded that the Hitachi 911 meets the requirements in terms of analytical performance, reliability, versatility and speed for an analyzer to be used in a routine (urine) setting, while having a distinct role in special (serum/whole blood) measurements.

Contribution of creatine kinase MB mass concentration at admission to early diagnosis of acute myocardial infarction.

OBJECTIVE: To assess the diagnostic value at admission of creatine kinase MB mass concentration, alone or in combination with electrocardiographic changes, in suspected myocardial infarction. DESIGN: Prospective study of all consecutive patients admitted within 12 hours after onset of chest pain to a coronary care unit for evaluation of suspected myocardial infarction. SETTING: Large regional hospital. PATIENTS: In 297 patients creatine kinase and creatine kinase MB activities and creatine kinase MB mass concentration were determined. Myocardial infarction according to the criteria of the World Health Organisation was diagnosed in 154 patients and excluded in 143 patients (including 70 with unstable angina pectoris). RESULTS: Sensitivity/specificity for creatine kinase MB mass concentration in patients admitted within 4 hours and 4-12 hours after onset of chest pain were 45%/94% and 76%/79% respectively. Corresponding values for creatine kinase activity were 20%/89% and 59%/83%, and for creatine kinase MB activity 16%/87% and 53%/87%. Raised creatine kinase MB mass concentration was seen in 17% of patients with unstable angina pectoris. Stepwise logistic regression analysis showed that independent predictors of acute myocardial infarction in patients admitted within 4 hours after onset of chest pain were electrocardiographic changes and creatine kinase MB mass concentration on admission; in patients admitted 4-12 hours after the onset of pain independent predictors were electrocardiographic changes and creatine kinase MB mass concentration and activity. CONCLUSION: Creatine kinase MB mass concentration is a more sensitive marker for myocardial infarction than the activity of creatine kinase and its MB isoenzyme. Electrocardiographic changes on admission in combination with creatine kinase MB mass concentration (instead of creatine kinase and creatine kinase MB activities) are best in diagnosing myocardial infarction.

Troponin T and myoglobin at admission: value of early diagnosis of acute myocardial infarction.

We studied the predictive power at admission of troponin T and myoglobin and compared them with that of CK and CK-MB activity and ECG in 290 consecutive patients admitted for evaluation of a suspected AMI. The likelihood ratio for an ischaemic ECG at admission < 4 h (between 4 and 12 h) after onset of chest pain was 2.85 (1.92), for a inconclusive ECG 1.53 (1.98) and for a normal ECG 0.27 (0.35). In patients admitted < 4 h after onset of chest pain, the likelihood ratio for abnormal and normal myoglobin concentrations (8.06 and 0.67) was considerably better for detection of AMI as defined by the WHO criteria than for the other markers, including the ECG. In patients admitted 4-12 h after onset of chest pain, the likelihood ratios for abnormal and normal myoglobin concentrations were 4.88 and 0.42; for troponin T 3.11 and 0.31; for CK activity 3.44 and 0.49 and for CK-MB activity 4.08 and 0.54 respectively. The sensitivity for troponin T (64%) was better than that of the other markers but its specificity (74%) was worse, because in patients with unstable angina troponin T was frequently elevated (37%). Stepwise logistic regression analysis showed that the best predictors of AMI within 4 h after onset of chest pain were the ECG and myoglobin and between 4-12 h after onset of chest pain the ECG, CK-MB activity and myoglobin.

The mass concentrations of serum troponin T and creatine kinase-MB are elevated before creatine kinase and creatine kinase-MB activities in acute myocardial infarction.

The time-related frequency of elevated results for the mass concentrations of the MB isoenzyme of creatine kinase and of troponin T were compared with that of creatine kinase and creatine kinase-MB activity in patients with acute myocardial infarction. Patients (322; 175 with and 147 without myocardial infarction) consecutively admitted for evaluation of possible acute myocardial infarction were investigated. Reference limits for troponin T (0.1 microgram/l) and creatine kinase-MB mass concentration (5.0 micrograms/l) were exceeded frequently in patients with unstable angina pectoris (troponin T 43%, creatine kinase-MB mass concentration 24%) in contrast to patients with no acute ischaemic heart disease (both < 5%). Within 4 and between 4-8 hours after onset of chest pain, the frequency of elevated results for creatine kinase-MB mass concentration and troponin T in patients with acute myocardial infarction was considerably higher (20-30%) than for creatine kinase and creatine kinase-MB activity. Creatine kinase-MB mass concentration and troponin T both allowed earlier diagnosis of acute myocardial infarction than creatine kinase and creatine kinase-MB activity, but troponin T was not elevated before the creatine kinase-MB mass concentration.

Failure of new biochemical markers to exclude acute myocardial infarction at admission.

In a substantial proportion of patients with suspected myocardial infarction, biochemical markers are needed for clinical decision-making at the time of admission, because electrocardiographic (ECG) recordings are inconclusive. We have assessed the usefulness for exclusion of myocardial infarction at admission of the newer markers creatine kinase MB (CK-MB) mass concentration, troponin T, and myoglobin in comparison with the routinely used markers creatine kinase (CK) and CK-MB activity. 290 consecutive patients were enrolled. Acute myocardial infarction was diagnosed on the basis of clinical history, ECG criteria, and time-dependent changes in CK and CK-MB activity. 153 patients had definite acute myocardial infarction. Troponin T had the highest sensitivity for prediction of acute myocardial infarction; high concentrations (above the upper reference limits) were found in 98 (64%) of the patients with infarctions compared with 92 (60%) for CK-MB mass concentration, 76 (50%) for myoglobin, 61 (40%) for CK activity, and 53 (35%) for CK-MB activity. However, troponin T also had the highest "false-positive" rate; of 137 patients without myocardial infarction, 36 (26%) had high troponin T concentrations. Sensitivity, specificity, and positive and negative predictive values were calculated in relation to time between onset of chest pain and hospital admission. Although CK-MB mass concentration was, by a small margin, the best marker in patients admitted within 8-10 h of onset of chest pain, all the markers had negative predictive values too low to allow exclusion of acute myocardial infarction at admission in patients with symptoms suggestive of myocardial infarction of less than 10 h duration.

Rapid determination of serum myoglobin with a routine chemistry analyzer.

A turbidimetric immunoassay system (Turbitime system, Behringwerke AG) allows rapid determination of myoglobin in serum. We adapted the reagents for this myoglobin assay (Turbiquant myoglobin) for use with a Hitachi 717 analyzer. No high-dose hook effect was observed up to 15,000 micrograms/L. Interassay CVs were 4.6% (mean = 72.0 micrograms/L; n = 9) and 2.5% (mean = 365.6 micrograms/L; n = 11). The calibration curve was stable for at least 1 month. Hemolysis did not interfere, and turbidity from lipemia interfered only when absorbance exceeded 2.0 A. Results of this method (y) correlated well with those by the Turbitime method (y = 1.256x - 44.1 micrograms/L; n = 91; r = 0.991) and by a commercially available radioimmunoassay (Byk-Sangtec; y = 0.739x - 42.2 micrograms/L; n = 94; r = 0.991). The upper limit (95th percentile) of the reference interval for myoglobin was estimated at 57.9 micrograms/L. The positive predictive value for results of myoglobin at admission was 89% with this upper reference limit and 99% with 100 micrograms/L, whereas the negative predictive value was about 60% for both limits.