title: Reproductive hormone reference intervals for healthy fertile young men: evaluation of automated platform assays.
abstract: BACKGROUND
  Management of male infertility and/or androgen deficiency requires accurate hormonal measurements with valid reference intervals.
  
  
  OBJECTIVE
  The objective of this study was to develop a valid reference panel of blood samples from healthy eugonadal young men with verified normal reproductive function and to use this panel to evaluate the performance of seven fully automated, commercial multiplex immunoassay platforms used to measure serum total testosterone (T), LH, and FSH.
  
  
  METHODS
  This was an observational study of consistency among seven different automated immunoassays for each of total T, LH, and FSH. Each method was implemented in two laboratories, with each repeating the analysis of the full reference panel samples twice. Serum T concentrations were also measured by gas chromatography/mass spectrometry (GC/MS), and serum inhibin B levels were determined by an ELISA.
  
  
  METHODS
  The study was performed at commercial, high-volume, clinical pathology laboratories.
  
  
  METHODS
  From 147 men screened, sera from 124 healthy, reproductively normal men (age, 21-35 yr) with normal sperm output were used as a reference panel. All laboratories selected for elite performance in the national immunoassay quality assurance program agreed to participate.
  
  
  METHODS
  For each of the 868 assays, descriptive statistics were calculated in the natural and log-transformed scales and were analyzed by nested, repeated measures ANOVA after log transformation. Reference intervals, defined as 95% confidence limits, were calculated using arithmetic (natural scale), geometric (log scale) and nonparametric methods.
  
  
  RESULTS
  Descriptive statistics and reference intervals for serum T, LH, and FSH differed widely and significantly between methods, but variation between laboratories for the same assay was negligible. All T methods showed significant differences in regression slope and intercept in deviance plots as well as in estimated reference ranges compared with the independent GC/MS reference method. Although similar between-method differences existed for gonadotropin assays, the smaller quantitative discrepancies allowed assignment of consensus reference intervals for serum FSH (1.3-8.4 IU/liter) and LH (1.6-8.0 IU/liter), although these differed from manufacturers' currently quoted expected values.
  
  
  CONCLUSIONS
  Using a reference panel of sera from healthy eugonadal young men with verified normal reproductive function, major differences exist between commercial T immunoassays as well as divergence from the GC/MS standard. This impairs their clinical diagnostic utility and requires substantial improvements in automated T immunoassay technologies or a switch to GC/MS methods. Gonadotropin assays showed less variability, but current high-throughput immunoassays remain suboptimal to confirm accurate diagnosis of azoospermia or androgen deficiency.
authors.affiliation: Royal College of Pathologists of Australasia and Australasian Association of Clinical Biochemists, Chemical Pathology Quality Assurance Programs Pty. Ltd., Flinders Medical Centre, South Australia 5042, Australia.
authors.forename: Ken
authors.initials: K
authors.lastname: Sikaris
authors.forename: Robert I
authors.initials: RI
authors.lastname: McLachlan
authors.forename: Rymantas
authors.initials: R
authors.lastname: Kazlauskas
authors.forename: David
authors.initials: D
authors.lastname: de Kretser
authors.forename: Carol A
authors.initials: CA
authors.lastname: Holden
authors.forename: David J
authors.initials: DJ
authors.lastname: Handelsman
mesh_terms: D000328:Adult; D005640:Follicle Stimulating Hormone / Q000097:blood*; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007265:Inhibins / Q000097:blood; D007986:Luteinizing Hormone / Q000097:blood*; D008297:Male; D012016:Reference Values; D013739:Testosterone / Q000097:blood*
chemical_list: C422364:inhibin B; D013739:Testosterone; D007265:Inhibins; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone