Please only refer to the current product lot insert enclosed with the kits package for execution

and reporting

.
0123 130206011M: 100 tests
130606011M: 050 tests

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MAGLUMI Direct Renin (CLIA)
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INTENDED USE
The kit is an in vitro chemiluminescent immunoassay for the quantitative determination of Renin in human EDTA-plasma
specimens using the MAGLUMI series Fully-auto chemiluminescence immunoassay analyzer (including Maglumi 600, Maglumi
800, Maglumi 1000, Maglumi 2000, Maglumi 2000 Plus, Maglumi 4000 and Maglumi 4000 Plus).
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SUMMARY AND EXPLANATION OF THE TEST

Renin, a polypeptidic enzyme (MW~ 40000) also known as angiotensinogenase, is a circulating protease secreted by
1.2
juxtaglomerular cells in the juxtaglomerular apparatus of the kidneys in response to low blood volume or low body NaCl content .
3
It is released in response to physiological stimuli like decreased blood volume and blood pressure , and sodium depletion. Renin
4-6
inactive precursor is prorenin, which is converted into renin by two steps . Firstly, prorenin undergoes a reversible
conformational change (which gives rise to activated prorenin); secondly, 46 amino acids of prorenin are proteolytically cleaved to
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produce active renin, a glycoprotein formed of 340 amino acid residues. Part of prorenin escapes proteolytic cleavage to renin
7
and is released into the circulation . Prorenin can be activated by different ways, like cryoactivation, acidification or partial
8,9 10
proteolysis . Prorenin secretion does not appear to be tightly regulated, whereas renin secretion is strictly controlled . Blood
concentration of prorenin is approximately ten-fold greater than that of renin. Renin is termed a double-domain enzyme, because
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11
the N- and C-terminal ends are quite similar. Each domain contains a single aspartic acid residue, critical for catalytic activity .
Renin activates the renin-angiotensin system by cleaving angiotensinogen produced in the liver into angiotensin I (inactive) which
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is further converted into angiotensin II (active) in the vascular epithelium of the lung . Angiotensin II can cause vasoconstriction
by stimulating the central nervous system, in addition it stimulates ADH (antidiuretic hormone) secretion and aldosterone
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13,14
secretion from the adrenal gland . Regulation of blood pressure and renal glomerular filtration control are the most important
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functions of renin -angiotensin system .

PRINCIPLE OF THE TEST

The Direct Renin assay is a sandwich chemiluminescence immunoassay.
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The sample (or calibrator/control, if applicable), magnetic miccrobeads coated with anti-Renin monoclonal antibody, and ABEI
labeled with another anti-Renin monoclonal antibody are mixed thoroughly and incubated at 37°C, formed sandwich
immuno-complexes. After precipitation in a magnetic field, the supernatant is decanted and then a wash cycle is performed.
Subsequently, the Starter 1+2 are added to initiate a chemiluminescent reaction. The light signal is measured by a photomultiplier
within 3 seconds as relative light units (RLUs), which is proportional to the concentration of Renin present in the sample (or
calibrator/control, if applicable).
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KIT COMPONENTS
Material Provided

100 tests 50 tests

Components Contents
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(REF: 130206011M) (REF: 130606011M)

Coated with anti-Renin monoclonal

Magnetic Microbeads 2.5 mL 2.0 mL
antibody, containing BSA, NaN3 (<0.1%).
Renin antigen (recombinant), containing
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Calibrator Low 2.0 mL 1.0 mL

BSA, NaN3 (<0.1%).
Renin antigen (recombinant), containing
Calibrator High 2.0 mL 1.0 mL
BSA, NaN3 (<0.1%).
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Labeled with anti-Renin monoclonal

ABEI Label 8.5 mL 5.5 mL
antibody, containing BSA, NaN3 (<0.1%).
Renin antigen (recombinant), containing
Control 1 2.0 mL 2.0 mL
BSA, NaN3 (<0.1%).
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Renin antigen (recombinant), containing

Control 2 2.0 mL 2.0 mL
BSA, NaN3 (<0.1%).
All reagents are provided ready-to-use.

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 Please only refer to the current product lot insert enclosed with the kits package for execution and reporting

Accessories Required But Not Provided

MAGLUMI Series:
Reaction Module REF: 630003
Starter 1+2 REF: 130299004M
Wash Concentrate REF: 130299005M
Light Check REF: 130299006M
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Please order accessories from Shenzhen New Industries Biomedical Engineering Co., Ltd. (SNIBE) or our authorized
representative.

CALIBRATION
Test of assay specific calibrators allows the RLU values to adjust the assigned master curve. To perform an accurate calibration, we
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have provided the calibrators checked by WHO International Standard 68/356. Results are determined via a calibration curve which
is instrument-specifically generated by 2-point calibration (10 calibrations) and a master curve provided via the reagent Radio
Frequency Identification (RFID) CHIP.
Recalibration is recommended if any of the following conditions occurs:
 After each exchange of lots (Reagent or Starter Reagents).

 Every week and/or each time a new reagent kit is used (recommended).
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 After instrument service is required.

 If controls lie outside the expected range.

 Whenever room temperature changes exceed 5° C (recommended).

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QUALITY CONTROL
Follow government regulations or accreditation requirements for quality control frequency.
Internal quality control is only applicable with MAGLUMI system. For instructions for use and target value refer to Direct Renin
(CLIA) Quality Control Information. User needs to judge results with their own standards and knowledge.
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For detailed information about entering quality control values, refer to the operating instructions of MAGLUMI series Fully-auto
chemiluminescence immunoassay analyzer.
To monitor system performance and chart trends, commercially available quality control materials are required. Treat all quality
control samples the same as patient samples.A satisfactory level of performance is achieved when analyte values obtained are
within the acceptable Control Range for the system or within your range, as determined by an appropriate internal laboratory
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quality control scheme. If the quality control results do not fall within the Expected Values or within the laboratory’s established
values, do not report results. Take the following actions:
 Verify that the materials are not expired.

 Verify that required maintenance was performed.

 Verify that the assay was performed according to the instructions for use.

 Rerun the assay with fresh quality control samples.

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 If necessary, contact your local technical support provider or distributor for assistance.

SPECIMEN COLLECTION AND PREPARATION

 When prorenin, the inactive precursor of renin, is cryoactivated to renin during sample handling, falsely elevated results are
obtained. Cryoactivation occurs when patient samples are chilled to temperatures of 4°C or below for extendedperiods of time,
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and when samples are chilled but still liquid (i.e., not frozen). Cryoactivation of prorenin to renin occurs more rapidly in serum.
Prorenin blood concentration is approximately ten-fold greater than that of renin.
 The only sample material validated is human EDTA-plasma. Use of serum, heparinized plasma, and citrated plasma samples
provides lower renin values, and is therefore not recommended.
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 Careful standardization of the patient preparation and sampling conditions is strongly recommended.Collect blood at room
temperature by venipuncture, in siliconized glass tubes, vacutainers (violet cap) or equivalent, containing EDTA as anticoagulant.
The presence of haemolysis may indicate mistreating during sample collection or handling.
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 Fasting specimens are recommended but not required. Record the time of day and the patient’s posture during blood collection
(supine, upright or seated).
 Do not pre-chill EDTA blood collection tubes nor store tubes on ice, but process blood at room temperature. Centrifuge tubes in a
non-refrigerated centrifuge, separate EDTA-plasma from cells immediately after centrifugation, then aliquot and deep-freeze at
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-20°C or below instantly.

 Carefully thaw before testing, mix the thawed samples and check for and remove air bubbles before assaying.
 Grossly haemolyzed or lipaemic samples as well as samples containing particulate matter or exhibiting obvious microbial
contamination should not be tested.

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 Do not use clotted samples.

 Avoid repeated freeze-thaw cycles. The EDTA-plasma samples can be frozen and thawed for only once, and may be stored up
to 60 days frozen at -20°C.
 It is recommended to test plasma samples immediately after loading on to the instrument.
 The minimum volume required for a single determination of Direct Renin is 100 µL.

WARNING AND PRECAUTIONS FOR USERS

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
 For In Vitro Diagnostic Use.
 Follow the package insert carefully. Reliability of assay results cannot be guaranteed if there are any deviations from the
instructions in this package insert.
Safety Precautions
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 CAUTION: This product requires the handling of human specimens. It is recommended that all human sourced materials be

considered potentially infectious and handled in accordance with the 29 CFR 1910.1030 Occupational exposure to bloodborne
pathogens. Biosafety Level 2 or other appropriate biosafety practices should be used for materials that contain or are suspected
of containing infectious agents.
 All samples, biological reagents and materials used in the assay should be considered potentially able to transmit infectious

agents. They should therefore be disposed of in accordance with the practices of your institution. Discard all materials in a safe
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and acceptable manner and in compliance with prevailing regulatory requirements.

 This product contains Sodium Azide. Dispose of contents and containers must be in accordance with all local, regional and

national regulations.
 Refer to safety data sheets which are available on request.
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Handling Precautions
 Do not use reagent kits beyond the expiration date.

 Do not interchange reagent components from different reagents or lots.

 Prior to loading the Reagent Kit on the system for the first time, the Reagent Kit requires mixing to re-suspend magnetic
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microbeads that have settled during shipment.

 For magnetic microbeads mixing instructions, refer to the Preparation of the Reagent section of this package insert.

 To avoid contamination, wear clean gloves when operating with a reagent kit and sample.

 Over time, residual liquids may dry on the septum surface. These are typically dried salts which have no effect on assay efficacy.

 For detailed discussion of handling precautions during system operation, refer to the SNIBE service information.
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Storage and Stability

 Sealed: Stored at 2-8° C until the expiration date.
 Opened at 2-8° C: Minimum stability is 4 weeks.
 On-board: Minimum stability is 4 weeks.

 To ensure the best kit performance, it is recommended to place opened kits in the refrigerator after the end of the intraday test
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work. It is still possible to keep on using the kit beyond the opened or on-board period if the controls are found within the
expected ranges.
 Keep upright for storage to facilitate later proper resuspension of magnetic microbeads.

 Keep away from sunlight.

TEST PROCEDURE
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Preparation of the Reagent

 Resuspension of the magnetic microbeads takes place automatically when the kit is loaded successfully, ensuring the magnetic

microbeads are totally resuspended homogenous prior to use.

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 To ensure proper test performance, strictly adhere to the operating instructions of MAGLUMI series Fully-auto

chemiluminescence immunoassay analyzer. Each test parameter is identified via a RFID CHIP on the Reagent. For further
information please refer to the operating instructions of MAGLUMI series Fully-auto chemiluminescence immunoassay analyzer.
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DILUTION
Dilutions are not generally needed due to the broad assay measuring range. In the rare instance when sample dilution is requested,
please refer to your laboratory procedures.
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High-Dose Hook
For the Direct Renin assay, no high dose hook effect was observed when samples containing Renin up to 150,000 μIU /mL.

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 Please only refer to the current product lot insert enclosed with the kits package for execution and reporting

LIMITATIONS
A skillful technique and strict adherence to the instructions are necessary to obtain reliable results.
Bacterial contamination or heat inactivation of the specimens may affect the test results.
A result within the expected range does not rule out the presence of disease and should be interpreted together with the patient’s
clinical picture and other diagnostic procedures.
Test results are reported quantitatively. However, diagnosis of a disease should not be based on the result of a single test, but
should be determined in conjunction with clinical findings in association with medical judgement.
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Any therapeutical decision should also be taken on a case-by-case basis.
Degradation of the molecule or prorenin cryoactivation may affect final results.
Patient samples containing human anti-mouse antibodies (HAMA) may give falsely elevated or decreased values. Although
HAMA-neutralizing agents are added, extremely high HAMA concentrations may occasionally influence results.

RESULTS
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Calculation of Results
The analyzer automatically calculates the Renin concentration in each sample by means of a calibration curve which is generated
by a 2-point calibration master curve procedure. The results are expressed in μIU/mL. For further information please refer to the
operating instructions of MAGLUMI series Fully-auto chemiluminescence immunoassay analyzer.
Interpretation of Results
The expected ranges for the Renin assay were obtained by testing 250 upright posture samples and 252 supine posture samples
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from healthy individuals in China and gave the following expected values:
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 Upright posture:4.20-45.6 μIU/mL (5 -95 percentiles).
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 Supine posture:3.11-41.2 μIU/mL (5 -95 percentiles).
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Blood was collected between 7:00 a.m. and 10:00 a.m. with the subjects either in an upright or supine position.
Upright samples were collected when individuals sat down to have their blood withdrawn, after standing for 30 minutes; supine
samples were collected after the individuals lay in supine position for at least 30 minutes.
Results may differ between laboratories due to variations in population and test method. It is recommended that each laboratory
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should establish its own reference range.

PERFORMANCE CHARACTERISTICS
Precision
Precision for the Direct Renin assay was determined as described in the CLSI EP5-A2, 2 controls, 4 human EDTA-plasma pools
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containing different concentration of analyte were assayed in duplicate of two at two independent runs per day for 20 testing days.
The result is summarized in the following table:
Within-Run Between-Run Total
Mean(µIU/mL)
Sample SD SD SD
(N=80) %CV %CV %CV
(µIU/mL) (µIU/mL) (µIU/mL)
EDTA-plasma Pool 1 4.498 0.355 7.89 0.082 1.82 0.365 8.11
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EDTA-plasma Pool 2 19.970 1.217 6.09 0.514 2.57 1.322 6.62

EDTA-plasma Pool 3 99.899 4.306 4.31 1.060 1.06 4.435 4.44
EDTA-plasma Pool 4 879.302 16.996 1.93 2.326 0.26 17.154 1.95
Control 1 40.047 2.076 5.18 0.715 1.79 2.198 5.49
Control 2 104.940 3.500 3.34 2.929 2.79 4.564 4.35
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Limit of Blank (LoB)

It is defined as the highest measurement result that is likely to be observed for a blank sample. The LoB for the Direct Renin assay is
0.5 µIU/mL.
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Limit of Detection (LoD)

The LoD for the Direct Renin assay is 1.0 µIU/mL.
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Measuring Range
0.5-1000 µIU/mL (defined by the limit of blank and the maximum of the master curve). Values below the limit of blank are reported
as <0.5 µIU/mL. Values above the measuring range are reported as >1000 µIU/mL.
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Linearity
The assay is linear between 1.0 µIU/mL and 1000 µIU/mL based on a study performed with guidance from CLSI EP6-A. Nine
equally distributed levels of samples were prepared by blending an EDTA-plasma sample containing Renin 1050 µIU/mL with an
EDTA-plasma sample depleted of Renin (0.0 µIU/mL).The mean sample recovery ranged from 90% to 110%.
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Method Comparison
A total of 177 samples in the range of 0.53 to 973.73 µIU/mL were tested by the Direct Renin assay and a commercially available
immunoassay according to CLSI EP-9-A3. The data from the resulting linear regressions are summarized as:
2
y=1.031x-3.089, r =0.991.

Analytical Specificity
The specificity date of the assay was obtained by adding these potential cross reactant to EDTA-plasma samples. The
Cross-reactivivity of the asssay was shown to be non-detectable for Beta2-microglobulin (<50 μg/mL), Cathepsin D (<1.5 μg/mL),
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Trypsin (<1.6 μg/mL), and Plasmin (<100 μg/mL).

Endogenous Interference
Substances up to the following concentrations did not interfere with the assay:
 Bilirubin 20 mg/dL
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 Triglyceride 3000 mg/dL

 Hemoglobin 500 mg/dL

REFERENCES
1. Toffelmire EB, Slater K, Corvol P, et al. Response of plasma prorenin and active renin to chronic and acute alterations of
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renin secretion in normal humans. Studies using a direct immunoradiometric assay. J Clin Invest. (1989) 83, 679–687.
2. Primary structure of the human Renin gene: Hardman J.A; and al. DNA (1984): 3(6):457-468.
3. Control of glomerular filtration rate by rennin-angiotensin system: Hall J. E.and al; Am.J.Physiol. (1977) :
233(5) :F366-F372.
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4. Navar LG, Inscho EW, Majid DSA, et al. Paracrine regulation of the renal microcirculation.Physiol. Rev. (1996) 76, 425–536.
5. Carey RM, Padia SH. Angiotensin AT2 receptors: control of renal sodium excretion and blood pressure. Trends Endocrinol
Metab. (2008) 19, 84-7.
6. Raised aldosterone to renine ratio predicts antihypertensive efficacity of spironolactone: a prospective cohort follow-up
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study: Lim P.O. and al; Br. J. Clin. Pharmacol. (1999): 48(5) :756-760 .
7. Müller MW, Todorov V, Krämer BK, Kurtz A. Angiotensin II inhibits renin gene transcription via the protein kinase C pathway.
Pflugers Arch. (2002) 444, 499-505.
8. Screening of primary aldosteronism: Schirpenbach C. and al; Best Pract. Res. Clin. Endocrinol. Metab. (2006) : 20(3) :
369-384 .
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9. Nicar MJ. Specimen processing and renin activity in plasma. Clin. Chem. (1992) 38, 598.
10. Koeppen BM, Stanton BA. Renal Physiology (4th ed.). Philadelphia, PA. Mosby Physiology Monograph Series, 2007.
11. Diagnostic procedure in renovascular hypertension: Distler A. and al.Clinical nephrology (1991): 36(4):174-180.
12. Circulating and tissue angiotensin systems: Campbell D.J. J. Clin. Invest. (1987):79(1):1-6.
13. Spät A, Hunyady L. Control of aldosterone secretion: a model for convergence in cellular signaling pathways.Physiol Rev.
(2004) 84, 489-539.
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14. Pitarresi TM. Rubattu S, Heinrikson R, Sealey JE. Reversible cryoactivation of recombinant human prorenin.J.Biol.Chem.
(1992) 267, 11753-9.
15. Nguyen G., Delarue F., Burcklé C., et al. Pivotal role of the renin/prorenin receptor in angiotensin II production and cellular
responses to renin. J Clin Invest. (2002) 109, 1417–1427.
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Shenzhen New Industries Biomedical Engineering Co., Ltd.

No.16, Jinhui Road, Pingshan New District, Shenzhen, 518122, P.R.China
Tel: 0086-755-21536601 Fax: 0086-755-28292740
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Lotus Global Co., Ltd.

1 Four Seasons Terrace, West Drayton, Middlesex London, UB7 9GG, United Kingdom
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Tel: 0044-20-75868010 Fax: 0044-20-79006187

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SYMBOLS EXPLANATIONS

Consult instructions for use Manufacturer

Temperature limit
Use-by date
( Store at 2-8 °C)
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Contains sufficient for Keep away from sunlight

Authorized representative in the

This way up
European Community
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In vitro diagnostic medical device Kit component

Catalogue number Batch code

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