A critical appraisal of accuracy and cost of laboratory methodologies for the diagnosis of hypogonadism: the role of free testosterone assays.

The biochemical diagnosis of male hypogonadism remains a controversial issue. The problem is compounded by the variety of laboratory assays available to measure serum testosterone (T) and the limited understanding, among clinicians, of their relative diagnostic validity. It is widely accepted that only the testosterone not bound to sex hormone-bounding globulin is metabolically active. Therefore, for diagnostic purposes it is frequent practice to order the measurement of free T (FT) or bioavailable T (BAT). Our objective is to describe the methods available for measuring FT and to review the literature to determine the relevance of ordering FT as a diagnostic laboratory tool in cases of suspected hypogonadism. We also provide our biochemical approach in evaluating men with T deficiency. The limited information available in this regard is frequently confined to the biochemistry literature. The few reliable studies indicate that analog-based measurement of FT offers no diagnostic or financial advantage over automated assay for total T. The manuscript also describes "How we do it." For optimal diagnostic accuracy and financial responsibility, total T and calculated FT (cFT) should be the tests employed for initial and confirmatory diagnosis respectively. Measurement of bioavailable T is an alternative option but not germane to the points to which we are calling attention in this paper. While clinicians should be discouraged from ordering FT assays, laboratories performing it should indicate what method was used and warned about possible reliability concerns. FT assays should no longer be a reimbursable test.

The significance of biological variation in the diagnosis of testosterone deficiency, and consideration of the relevance of total, free and bioavailable testosterone determinations.

PURPOSE: The diagnosis of testosterone deficiency syndrome is based on clinical manifestations and documentation of low testosterone. Which biochemical tests to use and the importance of morning sampling are still controversial. Biological variation (including interindividual and intraindividual biological variation) must be considered when interpreting individual results as it determines the usefulness of reference intervals and the change (reference change value) necessary for a significant difference between results. MATERIALS AND METHODS: A total of 87 healthy men (50 to 70+ years old) provided blood in the morning of the first day, and 4 weeks later in the morning and afternoon. Samples were frozen (-70C) and analyzed in the same run for serum testosterone, sex hormone-binding globulin and albumin, and bioavailable testosterone and free testosterone were calculated. RESULTS: Serum testosterone was lower in the afternoon by 1.5 nmol/l (43 ng/dl, p <0.05), with larger changes observed with higher morning values. However, this diurnal effect was dwarfed by the normal biological variation observed for repeat morning samples (serum testosterone +/- 4 nmol/l [115 ng/dl]). Between day intra-individual biological variation for morning serum testosterone was 18.7% while within day intra-individual biological variation was 12.9%. A change of 52% (reference change value) is necessary between serial morning results to confirm a significant difference. The biological variation parameters of calculated bioavailable testosterone and calculated free testosterone confer no advantage over total testosterone. CONCLUSIONS: Marked individuality of serum testosterone is evident even in healthy men. Because intraindividual biological variation is less than interindividual biological variation, reference intervals are marginally useful. The homeostatic set point of a patient could decrease by half and still be within the reference interval. Prospective establishment of an individual's baseline over repeated measurements or symptoms regardless of serum testosterone concentration should be used to guide clinical decisions.

Medical students' perceptions of an undergraduate research elective.

Recent years have seen a steady decline in the number of new physician-investigators (Association of American Medical Colleges, 2000). To encourage medical students to select research careers, the Queen's University Faculty of Health Sciences curriculum includes a mandatory Critical Enquiry elective in the 2nd year. An anonymous written survey was administered to medical students before and after the elective to determine their perceptions of the value of the elective and its impact on their decision to pursue a career in medical research. There was a significant increase in the number of students expressing an interest in pursuing a research career following the elective (35-42%, p = 0.029). Students recognized other benefits including the development of critical appraisal, information literacy, and critical thinking skills; and the opportunity to select an area of and form contacts for postgraduate training. Even students who choose not to pursue careers in medical research perceive benefits to a mandatory undergraduate research elective.

Salivary cortisol on ROCHE Elecsys immunoassay system: pilot biological variation studies.

OBJECTIVES: Salivary analysis is a noninvasive alternative that may be more acceptable to patients, especially children. As such, it has the potential for incorporation into comprehensive, dynamic investigations of metabolic dysfunctions - a significant advancement over a single time point serum analysis. In this study, we wanted to determine if the serum cortisol assay on our routine immunoassay analyzer could reliably measure salivary cortisol concentrations. Because of potential fluctuations in salivary concentrations, we included a biologic variation study as a main facet of our preliminary method evaluation. DESIGN AND METHODS: Twenty-eight healthy individuals (12 males, 16 females) volunteered to provide 5 nonconsecutive first morning saliva samples over a two-week period. Samples were stored frozen at home until the completion of the study. Following thawing and centrifugation, cortisol was measured in batch mode for each set of participant samples on the ROCHE Elecsys. Biologic variation was determined following removal of outliers. A method comparison was performed with the DPC Coat-A-Count Cortisol assay following the recommended modifications for salivary analysis, and with the Salimetrics HS-Cortisol two-site monoclonal assay optimized for salivary cortisol. RESULTS: Mean salivary cortisol concentration was 20.4 nmol/L. Analytical variation (CV(A) = 3.8%), within-subject variation (CV(I) = 6.3%), between-subject variation (CV(G) = 20.5%), index of individuality (II = 0.36) and reference change value (RCV = 20.4%) were determined. A negative 40% proportional bias was observed on the Elecsys compared to the two methods that have already been optimized for salivary analysis. CONCLUSIONS: This study demonstrated that salivary cortisol can be reliably measured on a routine automated immunoassay analyzer such as the ROCHE Elecsys. This particular assay needs to be optimized at the low end of the standard curve for routine use with salivary samples. Based on the relatively small intra-individual variation and low index of individuality, reference change values are preferable to the use of population reference intervals for this assay.