Detection and interpretation of lysergic acid diethylamide results by immunoassay screening of urine in various testing groups.

A total of 2259 urine samples were assayed for lysergic acid diethylamide (LSD) using radioimmunoassay (RIA, Coat-a-Count, Diagnostics Products) and a premarket cloned enzyme donor immunoassay (CEDIA, Boehringer Mannheim). Urine samples were obtained from patients admitted to the emergency room, patients in drug rehabilitation programs, and adults and juveniles in criminal probation programs. An overall incidence of positive results was 0.80% for CEDIA (500-pg/mL cutoff) and 0.89% and 0.18% for RIA at cutoffs of 250 and 500 pg/mL, respectively. Of the CEDIA-positive samples, only 17 and 11% were positive by RIA at 250 and 500 pg/mL, respectively, whereas among RIA-positive samples, only 10% of those > 250 pg/mL and only 25% of those > 500 pg/mL were positive by CEDIA. Moreover, only 2 of 25 of samples positive by one of these screening assays were confirmed by gas chromatography-mass spectrometry (GC-MS). It is likely that discrepancies in results between immunoassays are due to differences in antibody specificities used to detect LSD metabolites. In addition, immunoassays may be more sensitive than GC-MS for detecting LSD use as current confirmation assays are targeted towards detection of the parent drug only. The interpretation of results for LSD analysis must be made with knowledge of the limitations for each assay.

Detection of iron-deficiency anemia in hospitalized patients by zinc protoporphyrin.

The diagnostic value of zinc protoporphyrin (ZPP) as an indicator of iron-deficient anemia (IDA) in hospitalized patients is assessed in this study. ZPP was measured using an AVIV hematofluorometer with a coefficient of variation (CV) less than 5% and a recovery of greater than 97%. A reference range of 53-70 mu mol/mol heme was determined for ZPP in non-anemic patients in a hospital population. Hospitalized patients (221) with low hemoglobin (< 120 g/l) were evaluated for their iron status. ZPP and other anemia tests were performed. Macrocytic patients with mean corpuscular volume (MCV) greater than 98 fl) were excluded from the study. Seventy-four microcytic patients (MCV < 80 fl) were determined as having IDA according to a diagnostic algorithm. A distribution study of these microcytic patients showed that there was a significant overlap of values between the IDA and non-IDA patients for all serum anemia tests. A receiver-operator curve analysis revealed that ZPP has a relatively high degree of diagnostic efficiency better than iron and ferritin for this patient population. At a cutoff value > 170 mu mol/mol heme, ZPP has a sensitivity of 93% and a specificity of 90%. In addition, ZPP is also elevated in normocytic patients (MCV = 80-98 fl) with low ferritin values, who may have iron depletion. From these data, it is proposed that ZPP may be used as a screening tool for IDA in hospitalized patients.

Immunoassays for serum and urine myoglobin: myoglobin clearance assessed as a risk factor for acute renal failure.

We compared four immunoassays for serum and urine myoglobin. Within-run CVs were 5-13%, with biases seen between assays. Myoglobin was stable for 1 month in serum and 12 days in urine when the pH was adjusted to between 8.0 and 9.5. Hemoglobin caused no interference. We assayed 91 pairs of serum and timed urine specimens from 41 patients admitted for acute trauma or rhabdomyolysis. Most were treated with mannitol and alkalinization. Upon initial presentations, 21 patients with either low serum myoglobin concentrations (< 400 micrograms/L) or high myoglobin clearances (> or = 4 mL/min) had normal creatinine clearances and no clinical evidence of renal disease. The remaining 20 had low myoglobin clearances. Seven were in rhabdomyolysis-induced acute renal failure, or subsequently developed this complication. We suggest that low myoglobin clearance may indicate a high risk for developing renal failure or may be an early marker for kidney dysfunction. Low myoglobin clearance may prove useful in indicating failure of prophylactic treatment to clear myoglobin.

Preliminary evaluation of an experimental clinical chemistry analyzer developed for space medicine.

An experimental clinical chemistry analyzer system was designed and built to demonstrate the feasibility of clinical chemistry as part of a medical-care system at NASA's planned space station Freedom. We report the performance of the experimental analyzer, called a medical development unit (MDU), for selected analytes in a laboratory setting in preparation for a preliminary clinical trial at patients' bedsides in an intensive-care unit. Within-run CVs ranged from 0.7% for sodium to 7.1% for phosphorus; day-to-day CVs ranged from 1.0% for chloride to 23.4% for calcium. Correlation of patients' blood sample analyses compared well with those by Ektachem E700 and other high-volume central laboratory analyzers (r ranged from 0.933 for creatine kinase MB isoenzyme to 0.997 for potassium), except for hemoglobin (r = 0.901) and calcium (r = 0.823). Although several CVs obtained in this study exceeded theoretical desired precision limits based on biological variations, performance was adequate for clinical laboratory diagnosis. We examined the effect of potentially interfering concentrations of hemoglobin, bilirubin, and lipids: the only effect was negative interference with calcium analyses by high concentrations of bilirubin. We also examined the effects of preanalytical variables and the performance of experimental sample-transfer cups designed to retain sample and reference liquid in microgravity. Continued development of the MDU system is recommended, especially automation of sample processing.

Creatine kinase MB isoforms in patients with skeletal muscle injury: ramifications for early detection of acute myocardial infarction.

We measured total creatine kinase (CK), CK-MB isoenzyme, and the MB isoforms in 202 serum and plasma samples from nine groups of patients and normal individuals: 39 with acute myocardial infarction (MI), divided according to time between the onset of chest pain and blood collection (1-6 h, 7-12 h, and 13-48 h); 26 with chest pain for whom an MI was ruled out, sampled at admission; 17 undergoing bypass surgery or cardiac catheterization, sampled within 6 h after either procedure; 17 with acute skeletal muscle injury, sampled within 8 h after injury; 30 marathon runners immediately after a race; 17 runners and other athletes > 12 h after training or a race; 12 with cerebral injury or seizures, sampled at admission; 8 with closed head injury, sampled at admission; and 38 normal subjects. CK-MB (relative index) and MB isoforms (MB2/MB1) were respectively increased in 15% and 75% of MI patients 1-6 h after onset, 94% and 94% after 7-12 h, and 88% and 8% after 12 h, and in 87% and 82% of cardiac surgery patients. MB isoforms were increased in most patients with acute skeletal muscle trauma and in subjects examined after exercise, but were within normal limits in patients for whom MI was ruled out, patients with cerebral trauma, and normal individuals. The relative index of MB/total CK was normal in essentially all individuals in the last groups, including those with acute skeletal muscle trauma. We concluded that the CK-MB isoform ratio is increased in both acute skeletal muscle injury and MI. The isoform ratio is most useful for distinguishing recent from old (> 12 h) injury.

Role of rapid immunoassays for urgent ("stat") determinations of creatine kinase isoenyme MB.

We compared the analytical performance of three immunoassays used to rapidly determine creatine kinase (EC 2.7.3.2; CK) isoenzyme MB in serum: Dade's "Stratus," Corning's "Magic Lite," and Hybritech's "Icon QSR CK-MB." Performance criteria included precision, analytical sensitivity, sample stability, and analytical and clinical correlation of results for serum samples taken from healthy individuals, patients with suspected and confirmed acute myocardial infarction, and patients after coronary artery bypass surgery. We also examined 31 samples taken from patients in the emergency room suspected of myocardial infarction, to evaluate the potential of these assays for early diagnosis. Although these assays differ in the manner in which CK-MB is measured, and therefore have different procedural requirements, we conclude that they are equivalent in overall assay performance. None of these assays, however, is sufficiently sensitive for early diagnosis of myocardial infarction; therefore, results cannot be used by cardiologists in deciding whether acute thrombolytic therapy should be given. Other management decisions, such as the optimal utilization of intensive-care bed space, may justify using these assays on a "stat" basis.

Early diagnosis of acute myocardial infarction by rapid analysis of creatine kinase isoenzyme-3 (CK-MM) sub-types.

We compared the clinical sensitivity, specificity, and diagnostic efficiency of measuring creatine kinase-3 (MM) isoenzyme sub-types (CK, EC 2.7.3.2) with the measurement of CK-2 (MB) isoenzymes for the diagnosis of acute myocardial infarction. Serial blood collections at 3-h intervals from 35 patients with acute myocardial infarction were examined. In attempts to reperfuse their coronary arteries, some of these patients were treated with pharmacological thrombolysis (streptokinase, tissue plasminogen activator), with or without coronary angioplasty. The infarction patients were divided into two groups: patients who were successfully treated with thrombolytic agents (i.e., they achieved coronary reperfusion), and patients who were treated unsuccessfully or who were not treated acutely. We also examined blood from 34 non-infarction patients. We measured CK-3 sub-types by both anion-exchange liquid chromatography and a modified high-voltage electrophoresis method, and CK-2 by immunoprecipitation. Our results show that during the first few critical 3 to 9 h after onset of chest pain, measurement of CK-3 sub-types has the highest diagnostic efficiency; in contrast, CK-2 has the highest efficiency during the 10- to 21-h time intervals. Thus early diagnosis of acute myocardial infarction can be based on rapid assays of CK-3 sub-types.

Measurement of creatine kinase MM sub-type by anion-exchange liquid chromatography.

Electrophoretic and isoelectric focusing studies have demonstrated the presence of multiple sub-forms of the major creatine kinase isoenzymes, and preliminary work has suggested that measurement of the mm sub-forms provides early diagnostic information concerning acute myocardial infarction. We developed an isocratic method based on liquid chromatography to examine further the clinical potential of measuring these sub-forms. An anion-exchange column is used coupled with the fluorometric detection of NADPH after reaction of the sub-forms with CK reagents, which are added through a post-column pump. We examined various chromatographic variables in producing the best separations. Results for blood collected from normal individuals and patients with acute myocardial infarction and skeletal muscle diseases are compared. We conclude that for routine CK-MM sub-form analysis, this chromatographic procedure is better suited than isoelectric focusing because of the faster turn-around and lower costs.

Comparison of enzyme immunoassay and immunoprecipitation for creatine kinase MB in diagnosis of acute myocardial infarction.

We compared the clinical performance of measuring creatine kinase (EC 2.7.3.2) isoenzyme MB by use of an enzyme immunoassay (Enzygnost CK-MB, Behring Diagnostics) with an immunoprecipitation method (Isomune-CK, Roche Diagnostics) for the diagnosis of acute myocardial infarction. Sera from 80 patients admitted to the coronary care unit because of chest pain were examined: 40 who had this diagnosis of myocardial infarction, and 40 in whom it was ruled out. In addition, sera from 40 apparently healthy individuals were examined. The clinical sensitivity and specificity of these methods were evaluated by use of receiver operating characteristic curves. We conclude that for clinical efficiency, this enzyme immunoassay is slightly superior to the immunoprecipitation assay we used, because of its greater analytical sensitivity and precision for measuring the mass of the isoenzyme.

Preparation of urine samples for liquid-chromatographic determination of catecholamines: bonded-phase phenylboronic acid, cation-exchange resin, and alumina adsorbents compared.

We compared results for the liquid-chromatographic determination of free norepinephrine and epinephrine in urine after purifying the catechols by the following methods: (a) acid-washed alumina, (b) weak cation-exchange resin (WCX), (c) a combination of weak cation-exchange resin followed by alumina (WCX-alumina), and (d) commercially available phenylboronic acid adsorbent. We evaluated analytical specificity, sensitivity, recovery, and turnaround time. The WCX-alumina combination produced the most sensitive and specific chromatograms for urinary catecholamines; the other methods took less processing time. Neither WCX nor alumina alone was suitable for routine work because of chromatographic interferences in a significant proportion of urines. The phenylboronic acid method is adequately sensitive and specific for norepinephrine and epinephrine, and samples can be assayed faster. Thus it provides a compromise between the high analytical performance of the WCX-alumina method and the speed of the WCX and alumina methods.