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automated electrochemiluminescence
immunoassay method (ECLIA)
Evaluation
of
analytical
performance
C. Acevedo,
M. Morini,
M. Gomez, M.E. Almagro, M.G. Ropelato
Laboratorio de Medicina S.A. Olaya 1644. Bs. As. Argentina. e-mail:
endocrinologia@labmedicina.com
BACKGROUND
The diagnosis and treatment of Cushings Syndrome represents a challenge in clinical endocrinology as the features of endogenous
hypercortisolism are common to other clinical conditions. The lack of circadian rhythm in adrenal cortical secretion is one of the
hallmarks of this disease. Salivary cortisol reflects the unbound, biologically active form of serum cortisol. Consequently, the
measurement of late evening salivary cortisol is especially useful for evaluating ambulatory patients and for the pediatric
population, given it is a noninvasive method
OBJECTIVE
To evaluate the analytical performance of the ECLIA method for the measurement of salivary cortisol
MATERIALS Y METHODS
An automated immunoassay with electrochemiluminescent signal (Modular E170 Roche ) was used
Allowable total error (TEa) was established as 3CVi = 20%
Samples of saliva were obtained from healthy volunteers (13 women and 7 men) between 26 and 45 years of age. BMI<25 kg/m .These were
collected at 8:00, 15:00 and 23:00 hours.
Precision: repeatability (CVr) and intermediate precision (CVi) of two levels of concentration were evaluated with pools of samples of saliva (P1
and P2) according to EP15-A2 CLSI protocol
P1: 8.3 nmol/l
P2: 16.6 nmol/l
Data were compared with manufacturers claims
The limit of quantification ( LoQ), defined as the lowest concentration that can be quantified with an intermediate precision, CVi <20%, was
determined based on Spenser et al. (Demmers LM and Spencer LA (eds.) National Academy of Clinical Biochemistry Support for the diagram and monitoring of thyroid disease. 2003,
13:4-126) Five concentrations of saliva near the LoQ claimed were run for 10 days
The reference interval (RI) of 8:00 am and 15:00 pm proposed by the manufacturer was ascertained following C28 A3 CLSI protocol. The 95
percentile was estimated for the 23:00 hour samples as this RI was not reported as product information
Linearity was verified across 0.74 17.00 nmol/L, an interval representing the lower zone of the measuring range ( 0.50 1750 nmol/l)
LINEARITY
RESULTS
Repeatability
P1 CVr
4.7%
4.0%
P2 CVr
6.4%
4.4%
Intermediate Precision
P1 CVi
10.0%
6.7%
P2 CVi
9.5%
5.6%
LoQ
Verified
Verified
Not Reported
Clinical Linearity
Women
Mujeres
12,00
9,00
6,00
3,00
0,00
Csol 8 hs
Csol 15 hs Csol 23 hs
Cort 8hs
Cort 15 hs
Cort 23hs
Varones
8:00 hours
Laboratorio de Medicina
12,00
Salivary en
Cortisol
nmol/L)
Cosrtisol
saliva ((nmol/L)
ROCHE
Men
9,00
6,00
3,00
0,00
Cort 15 hs
Cort 23hs
CONCLUSIONS
The assessment of the ECLIA analytical performance enabled us to verify the precision and reference interval
specifications for salivary cortisol claimed by the manufacturer .
The LoQ obtained allowed us to ensure an adequate performance in the clinical decision levels.
FUTURE OBJECTIVES
Confirming our preliminary results of the reference interval in the 23:00 hour samples will be of great value in the
evaluation of the hypercortisolism disorders.
Our mayor goal is to establish the laboratorys pediatric RI for salivary cortisol.