ms_11972103122V26.

Elecsys PTH
MODULAR ANALYTICS E170
cobas e 411
11972103 122 100
cobas e 601
cobas e 602

English ▪ 2nd incubation: After addition of streptavidin-coated microparticles, the

complex becomes bound to the solid phase via interaction of biotin and
System information streptavidin.
For cobas e 411 analyzer: test number 680
For MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 ▪ The reaction mixture is aspirated into the measuring cell where the
analyzers: Application Code Number 126 microparticles are magnetically captured onto the surface of the
electrode. Unbound substances are then removed with
Intended use ProCell/ProCell M. Application of a voltage to the electrode then induces
Immunoassay for the in vitro quantitative determination of intact parathyroid chemiluminescent emission which is measured by a photomultiplier.
hormone in human serum and plasma for the differential diagnosis of ▪ Results are determined via a calibration curve which is instrument-
hypercalcemia and hypocalcemia. This assay can be used intraoperatively. specifically generated by 2‑point calibration and a master curve provided
The electrochemiluminescence immunoassay “ECLIA” is intended for use via the reagent barcode or e‑barcode.
on Elecsys and cobas e immunoassay analyzers. Reagents - working solutions
Summary The reagent rackpack is labeled as PTH.
Parathyroid hormone (PTH) is a single-chain 84‑amino-acid peptide
produced by the parathyroid glands in response to decreased extracellular M Streptavidin-coated microparticles (transparent cap), 1 bottle, 6.5 mL:
concentrations of ionized calcium. Its main role is to increase serum Streptavidin-coated microparticles 0.72 mg/mL; preservative.
calcium levels by stimulating the release of calcium from bone and its renal
re-absorption in the distal tubule. In the proximal tubule, PTH stimulates the R1 Anti-PTH-Ab~biotin (gray cap), 1 bottle, 7 mL:
synthesis of calcitriol which in turn increases intestinal absorption of calcium
and exerts an endocrine feed-back on the secretion of PTH at the Biotinylated monoclonal anti‑PTH antibody (mouse) 2.3 mg/L;
parathyroid level. PTH also decreases the renal re-absorption of phosphate phosphate buffer 100 mmol/L, pH 7.0; preservative.
in the proximal tubule, thereby decreasing serum phosphate.1 R2 Anti-PTH-Ab~Ru(bpy) (black cap), 1 bottle, 7 mL:
Parathyroid gland disorders lead to elevated or depressed blood calcium
levels (hypercalcemia or hypocalcemia) brought about by a change in the Monoclonal anti‑PTH antibody (mouse) labeled with ruthenium
secretion of PTH. complex 2.0 mg/L; phosphate buffer 100 mmol/L, pH 7.0;
Detection of subfunctioning parathyroid glands (hypoparathyroidism) preservative.
requires the use of a highly sensitive test in order to be able to measure
PTH levels well below normal. Hyperfunctioning of the parathyroid glands Precautions and warnings
results in an increased secretion of PTH (hyperparathyroidism). Primary For in vitro diagnostic use.
causes are adenomas of the parathyroid glands. In secondary Exercise the normal precautions required for handling all laboratory
hyperparathyroidism the blood calcium level is low as a result of other reagents.
pathological states (e.g. vitamin D deficiency).2 Disposal of all waste material should be in accordance with local guidelines.
The determination of PTH intraoperatively during adenoma resection in the Safety data sheet available for professional user on request.
parathyroid glands has been reported for primary hyperparathyroidism,3,4 Avoid foam formation in all reagents and sample types (specimens,
secondary hyperparathyroidism relating to renal failure,5,6 and tertiary calibrators and controls).
hyperparathyroidism post renal transplant surgery.7 Because PTH has a Reagent handling
reported half‑life of 3‑5 minutes,8 a significant drop in PTH levels after
resection of the abnormal gland or glands enables the surgeon to assess The reagents in the kit have been assembled into a ready‑for‑use unit that
whether all hyperfunctioning parathyroid tissue has been removed from the cannot be separated.
patient.9 All information required for correct operation is read in from the respective
The National Academy of Clinical Biochemistry recommends routine use of reagent barcodes.
intraoperative PTH testing for patients undergoing surgery for primary Storage and stability
hyperparathyroidism, both in initial surgeries and in reoperative Store at 2‑8 °C.
procedures.10
Do not freeze.
The Kidney Disease Outcomes Quality Initiative (KDOQI) and Kidney
Disease Improving Global Outcomes (KDIGO) guidelines recommend that Store the Elecsys reagent kit upright in order to ensure complete
serum PTH concentration should be measured regularly in patients with availability of the microparticles during automatic mixing prior to use.
Chronic Kidney Disease (CKD) and maintained within the target ranges that
are defined according to the stage of CKD.11,12 Stability:
The Elecsys assay for determining intact PTH employs a sandwich test unopened at 2‑8 °C up to the stated expiration date
principle in which a biotinylated monoclonal antibody reacts with the after opening at 2‑8 °C 12 weeks
N‑terminal fragment (1‑37) and a monoclonal antibody labeled with a
ruthenium complexa) reacts with the C‑terminal fragment (38‑84). on the analyzers 8 weeks
The antibodies used in this assay are reactive with epitopes in the amino
acid regions 26‑32 and 37‑42. Specimen collection and preparation
Only the specimens listed below were tested and found acceptable.
a) Tris(2,2'-bipyridyl)ruthenium(II)-complex (Ru(bpy) )
Serum collected using standard sampling tubes.
Test principle
Sandwich principle. Total duration of assay: 18 minutes. K2‑EDTA and K3‑EDTA plasma.
Because of the short half‑life of PTH, it is recommended that, when serum
▪ 1st incubation: 50 µL of sample, a biotinylated monoclonal PTH‑specific is needed, the blood should be centrifuged immediately.
antibody, and monoclonal PTH‑specific antibody labeled with a
ruthenium complex form a sandwich complex. Preference should be given to EDTA plasma, as it is stable longer than
serum.

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Elecsys PTH
Criterion: Method comparison serum versus plasma, slope 0.9‑1.1 + exceptional cases the barcode cannot be read, enter the 15‑digit sequence
intercept within < ± 2 x analytical sensitivity (LDL) + coefficient of of numbers (except for the cobas e 602 analyzer).
correlation > 0.95. MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 analyzers:
Serum: Stable for 8 hours at 15‑25 °C, 2 days at 2‑8 °C, 6 months at PreClean M solution is necessary.
‑20 °C (± 5 °C). Bring the cooled reagents to approximately 20 °C and place on the reagent
Plasma: Stable for 2 days at 15‑25 °C, 3 days at 2‑8 °C, 6 months at ‑20 °C disk (20 °C) of the analyzer. Avoid foam formation. The system
(± 5 °C). automatically regulates the temperature of the reagents and the
The sample types listed were tested with a selection of sample collection opening/closing of the bottles.
tubes that were commercially available at the time of testing, i.e. not all Calibration
available tubes of all manufacturers were tested. Sample collection systems Traceability: This method has been standardized against a commercial PTH
from various manufacturers may contain differing materials which could test (RIA). The recovery of the NIBSC 95/646 (WHO) standard was
affect the test results in some cases. When processing samples in primary assessed by testing dilutions in human serum covering the measuring
tubes (sample collection systems), follow the instructions of the tube range (40‑4000 pg/mL) on 16 analyzers (cobas e 411 and cobas e 601
manufacturer. analyzers). The mean recovery was 100 % ± 4 %.
Centrifuge samples containing precipitates before performing the assay. Every Elecsys reagent set has a barcoded label containing specific
Do not use samples and controls stabilized with azide. information for calibration of the particular reagent lot. The predefined
Ensure the samples, calibrators and controls are at 20‑25 °C prior to master curve is adapted to the analyzer using the relevant CalSet.
measurement. Calibration frequency: Calibration must be performed once per reagent lot
Due to possible evaporation effects, samples, calibrators and controls on using fresh reagent (i.e. not more than 24 hours since the reagent kit was
the analyzers should be analyzed/measured within 2 hours. registered on the analyzer).
Materials provided Calibration interval may be extended based on acceptable verification of
calibration by the laboratory.
See “Reagents – working solutions” section for reagents.
Renewed calibration is recommended as follows:
Materials required (but not provided)
▪ after 12 weeks when using the same reagent lot
▪ 11972219122, PTH CalSet, for 4 x 1.0 mL
▪ after 7 days (when using the same reagent kit on the analyzer)
▪ 08243875190, CalSet PTH, for 4 x 1.0 mL
▪ as required: e.g. quality control findings outside the defined limits
▪ 05618860190, PreciControl Varia, for 4 x 3 mL
Quality control
▪ General laboratory equipment For quality control, use PreciControl Varia.
▪ MODULAR ANALYTICS E170 or cobas e analyzer In addition, other suitable control material can be used.
Accessories for cobas e 411 analyzer: Controls for the various concentration ranges should be run individually at
▪ 11662988122, ProCell, 6 x 380 mL system buffer least once every 24 hours when the test is in use, once per reagent kit, and
following each calibration.
▪ 11662970122, CleanCell, 6 x 380 mL measuring cell cleaning
solution The control intervals and limits should be adapted to each laboratory’s
individual requirements. Values obtained should fall within the defined
▪ 11930346122, Elecsys SysWash, 1 x 500 mL washwater additive limits. Each laboratory should establish corrective measures to be taken if
▪ 11933159001, Adapter for SysClean values fall outside the defined limits.
▪ 11706802001, AssayCup, 60 x 60 reaction cups If necessary, repeat the measurement of the samples concerned.
▪ 11706799001, AssayTip, 30 x 120 pipette tips Follow the applicable government regulations and local guidelines for
quality control.
▪ 11800507001, Clean‑Liner
Accessories for MODULAR ANALYTICS E170, cobas e 601 and Calculation
cobas e 602 analyzers: The analyzer automatically calculates the analyte concentration of each
sample (either in pg/mL or pmol/L).
▪ 04880340190, ProCell M, 2 x 2 L system buffer
▪ 04880293190, CleanCell M, 2 x 2 L measuring cell cleaning Conversion factors: pg/mL x 0.106 = pmol/L
solution pmol/L x 9.43 = pg/mL
▪ 03023141001, PC/CC‑Cups, 12 cups to prewarm ProCell M and
CleanCell M before use Limitations - interference
The assay is unaffected by icterus (bilirubin < 1112 µmol/L or < 65 mg/dL),
▪ 03005712190, ProbeWash M, 12 x 70 mL cleaning solution for run lipemia (Intralipid < 1500 mg/dL) and biotin (< 205 nmol/L or < 50 ng/mL).
finalization and rinsing during reagent change
The assay is affected by hemolysis ≥ 0.15 g/dL. Do not analyze samples
▪ 03004899190, PreClean M, 5 x 600 mL detection cleaning solution that show visible signs of hemolysis.
▪ 12102137001, AssayTip/AssayCup, 48 magazines x 84 reaction Criterion: Recovery within ± 10 % of initial value.
cups or pipette tips, waste bags
Samples should not be taken from patients receiving therapy with high
▪ 03023150001, WasteLiner, waste bags biotin doses (i.e. > 5 mg/day) until at least 8 hours following the last biotin
▪ 03027651001, SysClean Adapter M administration.
Accessories for all analyzers: No interference was observed from rheumatoid factors up to a
concentration of 1500 IU/mL.
▪ 11298500316, ISE Cleaning Solution/Elecsys SysClean,
5 x 100 mL system cleaning solution There is no high-dose hook effect at PTH concentrations up to
17000 pg/mL (1802 pmol/L).
Assay In vitro tests were performed on 16 commonly used pharmaceuticals. No
For optimum performance of the assay follow the directions given in this interference with the assay was found.
document for the analyzer concerned. Refer to the appropriate operator’s
manual for analyzer‑specific assay instructions. In rare cases, interference due to extremely high titers of antibodies to
analyte‑specific antibodies, streptavidin or ruthenium can occur. These
Resuspension of the microparticles takes place automatically prior to use. effects are minimized by suitable test design.
Read in the test‑specific parameters via the reagent barcode. If in

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For diagnostic purposes, the results should always be assessed in Precision was determined using Elecsys reagents and controls in a
conjunction with the patient’s medical history, clinical examination and other separate study according to protocol EP5‑A2 of the CLSI (Clinical and
findings. Laboratory Standards Institute): 2 runs per day in duplicate each for
21 days (n = 84). The following results were obtained:
Limits and ranges
Measuring range cobas e 411 analyzer
1.20‑5000 pg/mL or 0.127‑530 pmol/L (defined by the lower detection limit Repeatability Intermediate
and the maximum of the master curve). Values below the lower detection
limit are reported as < 1.20 pg/mL (< 0.127 pmol/L). Values above the precision
measuring range are reported as > 5000 pg/mL (> 530 pmol/L). Sample Mean SD CV SD CV
Lower limits of measurement pg/mL pmol/L pg/mL pmol/L % pg/mL pmol/L %
Lower detection limit of the test
PCc) 54.6 5.79 0.657 0.070 1.2 1.11 0.118 2.0
Lower detection limit: 1.20 pg/mL (0.127 pmol/L) Varia 1
The Lower Detection Limit represents the lowest measurable analyte level
that can be distinguished from zero. It is calculated as the value lying two PC 182 19.3 2.43 0.258 1.3 3.14 0.333 1.7
standard deviations above that of the lowest standard (master calibrator, Varia 2
standard 1 + 2 SD, repeatability study, n = 21). c) PC = PreciControl
Dilution
MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 analyzers
Not necessary due to the broad measuring range.
Repeatability Intermediate
Expected values
precision
15‑65 pg/mL (1.6‑6.9 pmol/L)13,14
Each laboratory should investigate the transferability of the expected values Sample Mean SD CV SD CV
to its own patient population and if necessary determine its own reference pg/mL pmol/L pg/mL pmol/L % pg/mL pmol/L %
ranges.
PC 54.7 5.80 0.943 0.100 1.7 0.977 0.104 1.8
Specific performance data Varia 1
Representative performance data on the analyzers are given below.
Results obtained in individual laboratories may differ. PC 184 19.5 2.65 0.281 1.4 3.11 0.330 1.7
Varia 2
Precision
Precision was determined using Elecsys reagents and pooled human sera Method comparison
in a modified protocol (EP5‑A) of the CLSI (Clinical and Laboratory A comparison of the Elecsys PTH assay (y) with a commercial PTH test (x)
Standards Institute): 6 times daily for 10 days (n = 60); repeatability on using clinical samples gave the following correlations (pg/mL):
MODULAR ANALYTICS E170 analyzer, n = 21. The following results were
obtained: Number of samples measured: 152

cobas e 411 analyzer Passing/Bablok15 Linear regression

Repeatability Intermediate y = 1.01x + 4.86 y = 0.83x + 19.4
precision τ = 0.886 r = 0.991
Sample Mean SD CV SD CV The sample concentrations were between 1.4 and 1880 pg/mL (0.15 and
199 pmol/L).
pg/mL pmol/L pg/mL pmol/L % pg/mL pmol/L %
Analytical specificity
HSb) 1 26.7 2.83 0.711 0.075 2.7 1.73 0.184 6.5
No cross-reactivities were found for: Osteocalcin, PTH fragment 1‑37,
HS 2 52.5 5.56 0.853 0.091 1.6 2.07 0.220 3.9 PTH‑related protein (1‑86), bone‑specific alkaline phosphatase, and
β‑CrossLaps.
HS 3 261 27.7 4.0 0.424 1.5 7.81 0.829 3.0
Functional sensitivity
b) HS = human serum
6.0 pg/mL (0.64 pmol/L)
MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 analyzers The functional sensitivity is the lowest analyte concentration that can be
Repeatability reproducibly measured with an intermediate precision CV of ≤ 20 %.
Sample Mean SD CV Clinical investigations in intraoperative use
In 2006, the National Academy of Clinical Biochemistry published their
pg/mL pmol/L pg/mL pmol/L % Laboratory Medicine Practice Guidelines for point of care testing, entitled
HS 1 21.9 2.32 0.44 0.05 2.0 Evidence Based Practice for Point of Care Testing.10 The guidelines
recommend the use of intraoperative parathyroid hormone testing 1) for
HS 2 35.0 3.71 0.43 0.05 1.2 patients undergoing surgery for hyperparathyroidism, especially in
HS 3 123 13.04 1.31 0.14 1.1 minimally invasive or directed procedures, 2) for patients undergoing
reoperation, and 3) as a replacement for traditional laboratory
measurements of PTH during venous localization in order to help the
MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 analyzers angiography team guide sampling.
Intermediate precision The guidelines further recommend for patients undergoing
Sample Mean SD CV parathyroidectomy for hyperparathyroidism that baseline samples be
obtained preoperation exploration and pre‑excision of the gland, and that
pg/mL pmol/L pg/mL pmol/L % post‑excision sampling be drawn at 5 and 10 minutes post resection with a
HS 1 23.2 2.46 0.79 0.08 3.4 50 % reduction in PTH concentrations from the highest baseline level. The
guidelines also caution that additional samples may be necessary.10
HS 2 80.9 8.58 2.01 0.21 2.5 PTH testing during parathyroid surgery was conducted by several groups of
HS 3 240 25.4 6.72 0.71 2.8 investigators using the Elecsys PTH immunoassay.4,5,6,7,16

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The overall sensitivity and specificity of the assay to demonstrate Symbols
successful surgery as defined by postoperative reduction of calcium levels Roche Diagnostics uses the following symbols and signs in addition to
was 99.6 % and 93.7 %, respectively. those listed in the ISO 15223‑1 standard (for USA: see
References https://usdiagnostics.roche.com for definition of symbols used):
1 Souberbielle JC, Roth H, Fouque D. Parathyroid hormone Contents of kit
measurement in CKD. Kidney Int 2010;77:93–100.
Analyzers/Instruments on which reagents can be used
2 Nussbaum S, Potts JT. Advances in Immunoassays for Parathyroid
Hormone. Clinical Applications to Skeletal Disorders of Bone and Reagent
Mineral Metabolism. In Bilezikian JP, Levine MA, Marcus R (eds). The
Parathyroids: Basic an Clinical Concepts. Raven Press, New York Calibrator
1994:157-169. Volume after reconstitution or mixing
3 Bergenfelz A, Nordén NE, Ahrén B. Intraoperative fall in plasma levels
of intact parathyroid hormone after removal of one enlarged parathyroid GTIN Global Trade Item Number
gland in hyperthyroid patients. Eur J Surg 1991;157:109-112.
COBAS, COBAS E, ELECSYS and PRECICONTROL are trademarks of Roche. INTRALIPID is a trademark of
4 Ohe MN, Santos RO, Kunii IS, et al. Usefulness of a rapid Fresenius Kabi AB.
immunometric assay for intra-operative parathyroid hormone All other product names and trademarks are the property of their respective owners.
measurements. Braz J Med Biol Res 2003;36(6):715-721. Additions, deletions or changes are indicated by a change bar in the margin.
© 2019, Roche Diagnostics
5 Seehofer D, Rayes N, Ulrich F, et al. Intra-operative measurement of
intact parathyroid hormone in renal hyperparathyroidism by an
inexpensive routine assay. Langenbecks Arch Surg
2001;386(6):440-443.
6 Seehofer D, Rayes N, Klupp J, et al. Predictive value of intact Roche Diagnostics GmbH, Sandhofer Strasse 116, D-68305 Mannheim
www.roche.com
parathyroid hormone measurement during surgery for renal
hyperparathyroidism. Langenbecks Arch Surg 2005;390(3):222-229.
7 Haustein SV, Mack E, Starling JR, et al. The role of intra-operative
parathyroid hormone testing in patients with tertiary
hyperparathyroidism after renal transplantation. Surgery
2005;138(6):1066-1071.
8 Maier GW, Kreis ME, Renn W, et al. Parathyroid hormone after
adenectomy for primary hyperparathyroidism: A study of peptide
hormone elimination kinetics in humans. Jour Clin Endocrinol Metab
1998;83(11):3853-3856.
9 Carter AB, Howanitz TJ. Intra-operative testing for parathyroid
hormone: a comprehensive review of the use of the assay and the
relevant literature. Arch Pathol Lab Med 2003;127:1424-1442.
10 Nichols JH, Christenson RH, Clarke W, et al. National Academy of
Clinical Biochemistry Laboratory Medicine Practice Guidelines:
Evidence Based Practice for Point of Care Testing. AACC Press:2006.
11 National Kidney Foundation. K/DOQI clinical practice guidelines for
bone metabolism and diseases in chronic kidney disease. Am J Kidney
Dis 2003;42:S1-201.
12 KDIGO – Kidney Disease Improving Global Outcomes KDIGO clinical
practice guideline for the diagnosis, evaluation, prevention, and
treatment of chronic kidney disease-mineral and bone disorder
(CKDMBD). Kidney International 2009;76(Suppl 113).
13 Blind E. Measurement of Intact Parathyroid Hormone by an Extracting
Two-Site Immunometric Assay. In: Schmidt-Gayk H, Armbruster FP,
Bouillon R, (eds). Calcium regulating hormones, vitamin D metabolites,
an cyclic AMP. Heidelberg: Springer 1990:151.
14 Thomas L. Parathyroid hormone (PTH). Clinical Laboratory Diagnosis.
TH-Books, Frankfurt. 1st english edition 1998:248-250.
15 Bablok W, Passing H, Bender R, et al. A general regression procedure
for method transformation. Application of linear regression procedures
for method comparison studies in clinical chemistry, Part III.
J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.
16 Ohe MN, Santos RO, Kunii IS, et al. Usefulness of intra-operative PTH
measurement in primary and secondary hyperparathyroidism:
experience with 109 patients. Arq Bras Endocrinol Metab
2006;50(5):869-875.
For further information, please refer to the appropriate operator’s manual for
the analyzer concerned, the respective application sheets, the product
information and the Method Sheets of all necessary components (if
available in your country).
A point (period/stop) is always used in this Method Sheet as the decimal
separator to mark the border between the integral and the fractional parts of
a decimal numeral. Separators for thousands are not used.

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