This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Certification No. 226ADAMX00197000**Revised: October 2020 (3rd edition)* Revised: January 2017 (2nd edition)

This package insert must be read carefully prior to use.
Tacrolimus assay kit(Classification No.: 30421000)

reported to cause adverse reactions such as infection,
renal failure, and central nervous system disorders.
Because there is considerable individual variation in
the blood level of tacrolimus, it is necessary tomeasure blood concentrations and adjust the dosage
depending on the condition of the patient. According
to the Drug Interview Form, the therapeutic range is
5–20 ng/mL after organ transplantation (or 10–20
ng/mL after bone marrow transplantation whengraft-versus-host disease is more likely to occur) and
10–15 ng/mL in patients with ulcerative colitis(reducing to 5–10 ng/mL after 2 weeks oftreatment). It is mentioned that adverse reactions
tend to occur in patients with persistently high blood
trough concentrations (> 20 ng/mL).1)

Nanopia TDM Tacrolimus
General Precautions
1. This product is for in vitro diagnostic use, andmust not be used for any other purposes.
2. Clinicians should make a comprehensive clinicaldecision based on assay results in conjunction
with clinical symptoms and other examination
results.
3. Please read carefully the "PRECAUTIONS"
section, particularly under "Effects on Laboratory
Tests" of the package insert of each drug, for
information on the influence to the assay resultthat medications administered to the patient have.
Please also read carefully the "2) Crossreactivity," under "2. Interfering Substances," inthe "Procedural Precautions" section, as well as"2. Precautions for Assessment" in the"Assessment of Assay Results" section, of thispackage insert.
4. This product should be used only as directed inthis package insert. Reliability of results cannot be
guaranteed if there are any deviations from the
instructions in this package insert.
5. If the reagent accidentally comes in contact witheyes and/or mouth, rinse immediately with ample
water as first aid, and consult the doctor if
required.
6. Carefully read the operating instructions for eachtype of automated analyzers prior to using thisproduct. Parameters for each type of analyzers areavailable, and can be requested from SEKISUIMEDICAL CO., LTD. if required.
7. Perform a quality control test prior to assay toensure accuracy.
8. TACR Antibody Solution 1 contains humanderived ingredients that have been confirmed tobe negative for HBs antigens, HIV antibodies, andHCV antibodies. However, handle this reagent
with great care in the same manner as the
samples, because it is potentially infectious.

Assay Principle
1. Assay Principle
Pretreat the whole blood sample with methanol
and the Pretreatment Solution, and subsequently
centrifuge. Use the supernatant thus obtained as
the sample. When a certain amount of antitacrolimus antibody is added and reacted with asample, consumption of the antibody depends on
its content in the sample. When tacrolimuscoated latex is added, residual anti-tacrolimus
antibody reacts with the latex and forms
aggregates.
Since the extent of aggregation depends on thetacrolimus concentration in the sample, thetacrolimus concentration can be determined bymeasuring aggregation as the absorbance.
Sample (tacrolimus) + Anti-tacrolimus antibody───▶ Antigen-antibody reaction
Unreacted anti-tacrolimus antibody +Tacrolimus-coated latex───▶ Aggregation by antigen-antibody reaction
2. Features
1) Liquid reagents, ready-to-use.
2) It can be measured using the automatedanalyzers commonly used.
＊＊

Procedural Precautions ＊1. Properties of Samples and Sampling Methods
1) Samples(1) Whole blood (EDTA-treated) is used as the
sample.
(2) Use methanol (HPLC grade) and thePretreatment Solution in the kit for
pretreatment of the sample.
2) Storage of samples(1) Samples may be stored for up to 7 days in arefrigerator. If samples cannot be measured
within 7 days of collection, store them at−20C or lower and measure them within 6months. Stored samples should be brought
to room temperature (15–30C) before use.
(2) Samples may be frozen and thawed up to

Description (Kit Components)
Component: Ingredients
TACR Antibody Solution 1:
Anti-tacrolimus rabbit
monoclonal antibody
TACR Latex Reagent 2:
Tacrolimus-coated latex
Pretreatment Solution 3
Intended Use
Measurement of tacrolimus in whole blood
Tacrolimus is a macrolide that has a strong
immunosuppressive effect by selectively inhibiting
the activation of T cells. Tacrolimus has been

1

twice.
3) Measure samples immediately after
pretreatment.
2. Interfering substances
1) Assay results are not affected by free bilirubin(up to 19.1 mg/dL), conjugated bilirubin (up to
21.9 mg/dL), formazin turbidity (up to 1620
FTU), or rheumatoid factors (up to 500 IU/mL).
2) Cross-reactivity
The following table summarizes the crossreactivity with the tacrolimus metabolites andother drugs.
(1) Tacrolimus metabolites
If each metabolite is at the concentration
shown in the table below when thetacrolimus concentration is approximately 5–
21 ng/mL, cross-reactivity is as follows.
Metabolite
M-I (13-O-demethyl
tacrolimus)
M-II (31-O-demethyl
tacrolimus)
M-III (15-O-demethyl
tacrolimus)
M-IV (12-hydroxytacrolimus)
M-VII(13,15-O-didemethyl
tacrolimus)
M-VII (13,15-Odidemethyl tacrolimus)+ M-VI (13,31-Odidemethyl tacrolimus)

Concentration
tested(ng/mL)

Crossreactivity
※
(%)

20

10.5

20

1.3

20

8.3

3.3
20

174.8
10.5

20

11.5

20

4.0

Cyclosporine A
Diazepam
Digitoxin
DigoxinDiltiazem
Disopyramide
Erythromycin
Ethosuximide
Everolimus
Famotidine
Fluconazole
Flucytosine
Furosemide
Ganciclovir
Gentamicin
Hydralazine
Hydrochlorothiazide

Ibuprofen
Itraconazole
Kanamycin
AsulfateKanamycin
BsulfateKetoconazole
Labetalol
Lidocaine
Lithium
Methylprednisolone
Metoclopramide
Minoxidil
Morphine sulfate
Mycophenolicacid(2) Other drugs
If each drug is at the concentration shown in
the table below when the tacrolimus
concentration is approximately 5 or 12
ng/mL, cross-reactivity is as follows. The
recovery rate is 91–109%.
Drug nameConcentration
tested(ng/mL)

Crossreactivity※(%)

Acetaminophen
Acyclovir
Allopurinol
Amikacin sulfate
Amphotericin B
Ampicillin
Atenolol
Azathioprine
Azithromycin
Bromocriptine
Carbamazepine
Cefazolin
Ceftriaxone
Cephalosporin C
Chlorpromazine
Chloramphenicol
Chlordiazepoxide
Cimetidine
Ciprofloxacin
Clarithromycin
Clonidine
Colchicine
Cortisone

200,000
1,000,000
50,000150,000
100,000
100,000
40,000100,000
5,000
8,000
120,000
150,000
500,000
100,000
50,000250,000
20,000100,000
7,400
5,000
100
90
1,200

0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.2
0.7
0.0

N-Acetylprocainamide

Nadolol
Naproxen
Nicardipine
Nicotine
Nifedipine
Penicillin G
Pentobarbital
Phenobarbital
Phenytoin
Prazosin
Prednisolone
Primidone
Probucol
Procainamide
Propranolol
Quinidine
Ranitidine
Rifampicin
Salicylic acid
Sirolimus
Spectinomycin
Streptomycin
Sulfamethoxazole
Theophylline
Ticlopidine
Tobramycin
Triamterene
Trimethoprim

2

10,00020,000100,000
10,00060,000100,000
200,000
300,000
100
10,000100,000
40,000100,000
1,000,000
120,000
100,000
40,000400,000
100,000
100,000

0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
–0.5
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0

100,000

0.0

100,000
17,100100,000
35,000100,000

0.0
0.0
0.0
0.0
0.0

100,000
60,000100,000
100,000

0.0
0.0
0.0
0.0

120,000
1,200
100,000
500
20,000100,000
100,000
100,000
150,000
100,000
100,000
100,000
100,000
600,000
100,000
40,000100,000
200,000
100,000
500,000
300
100,000
100,000
150,000
250,000
150,000
100,000
100,000
40,000

0.0
0.0
0.0
0.1
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.2
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0

procedure is indicated below.

Valproic acid
500,000
0.0
Vancomycin
100,000
0.0
Verapamil
100,000
0.0※
Based on the Clinical and Laboratory
Standards Institute (CLSI) EP7-A2, crossreactivity was calculated by the following
equation.
Cross-reactivity (%) = ({[Concentration oftacrolimus in the sample after adding themetabolite or drug] – [Concentration oftacrolimus in the sample without the
metabolite or concomitant drug]} /[Concentration of the metabolite or drug inthe sample]) × 100
3. Others
1) Use Tacrolimus Calibrator for Nanopia as thecalibration material after carrying out
pretreatment in the same manner as for samples.
2) Precautions for assay range
If the tacrolimus concentration in the sample
exceeds the measurement range, dilute thesample with Tacrolimus Calibrator A, and thenperform pretreatment and re-measurement.

Sample +10 μL

37C Reagent 37C Measurement
Reagent(2)(1) Approx. 75 μL Approx.(Absorbance※
125 μL 5 min
3 min I )37C Measurement(AbsorbanceApprox. ※
II )
2 min
Calculation ofconcentration

Absorbance I and II: Absorbance at 700 nm※

Calibration material: Tacrolimus Calibrator(Manufacture's assigned value)
Assessment of Assay Results ＊＊1. Reference standard range
The therapeutic range of the whole bloodconcentration of tacrolimus is specified as 5–20
ng/mL after organ transplantation (or 10–20
ng/mL after bone marrow transplantation whengraft-versus-host disease is more likely to occur)and 10–15 ng/mL in patients with ulcerative
colitis (reducing to 5–10 ng/mL after 2 weeks oftreatment).1) However, the therapeutic range may
overlap the toxic concentration range. Therefore,
interpretation of assay results should also bebased on the patient's clinical findings and otherexamination results.
It is recommended that the blood concentrationof tacrolimus should be measured in a sufficientnumber of samples for statistical analysis andthat its therapeutic range should be determinedby each medical institution.
2. Precautions for Assessment
There may be reactions or interfering reactions
with non-target substances. If assay resultsappear to be unreliable, repeat the measurement(if necessary, after dilution) or try anotheranalytical methods.

Dosage/Administration (Assay Procedure)
1. Preparation of reagents
Reagent (1): TACR Antibody Solution 1 is readyto use.
Reagent (2): TACR Latex Reagent 2 is ready touse.
Pretreatment Solution: Pretreatment Solution 3 isready to use.
Before using this product, gently invert the TACR
Latex Reagent 2 bottle to mix it thoroughly, andcheck that there are no bubbles.
2. Instruments and Reagents
1) Micropipettes (10–100 μL and 100–500 μL)
2) Pipette tip3) Methanol (HPLC grade)4) Microcentrifuge tube
5) Vortex mixer
6) High-speed shaker (if used)
7) Microcentrifuge
3. Pretreatment of Samples
1) Gently mix the Pretreatment Solution andmethanol (HPLC grade) at a volume ratio of
1:4 in a clean and dry air-tight bottle, and use
this solution as the extraction solution (e.g.,mix 10 mL of the Pretreatment Solution with
40 mL of methanol [HPLC grade]).
The extraction solution can be used for 2
weeks from the day of preparation when kept
at room temperature.
2) Pipette 200 μL of the sample into a
microcentrifuge tube.
3) Add 200 μL of the extraction solution andmix for 15–30 seconds in a Vortex mixer orfor 5 minutes in a high-speed shaker.
4) After the microcentrifuge tube is let stand for
5 minutes at room temperature, perform
centrifugation (13000 rpm for 5 minutes).
5)
Dispense 200 μL of the supernatant thus
obtained, and use it as the sample.
4. Assay Procedure
This product is compatible with various types ofautomated analyzer. An example of the assay

Performance
1. Sensitivity(1) Absorbance of standard solution (0.0 µg/mL)is ≥ 850 (Abs. × 10000).
(2) The ratio of absorbance between standard
solution (0.0 ng/mL) and another standard
solution (approx. 2.0 ng/mL) is ≤ 88%.
2. Accuracy: 80–120 % of the expected assay value
3. Within-run Reproducibility:
Coefficient of variation ≤ 15 %(Test methods used for 1.–3. are in-housemethods.)
4. Measurement Range2): (On Hitachi 7180
automated analyzer)
1.5–30.0 ng/mL
5. Correlation2)
1) Whole blood (EDTA)
N=168
r=0.932
y=1.03x+0.71
Control method: Approved in vitro diagnostic(chemiluminescence immunoassay)
2) Whole blood (EDTA)
N=93
r=0.926y=1.06x+1.9
Control method: Approved in vitro diagnostic3

(enzyme immunoassay)
6. Standard Material
Tacrolimus (in-house standard material)

Storage and Shelf Life
1. Storage temperature: 2–8C
2. Shelf life: 1 year from the date of manufacture(The expiration date is printed on the outer
package.)

Precautions for Use or Handling *1. Precautions for Handling (to Ensure Safety)
1) All samples used in the test should be handled
as a material possibly infected with HIV, HBV,HCV, or other viruses. To prevent infection,use disposable gloves and avoid mouth
pipetting during the test.
2) Sodium azide is added as an antiseptic agent in
the TACR Antibody Solution 1 and TACR
Latex Reagent 2. Therefore, if the reagent
comes in accidentally contact with eyes, mouthor skin, rinse immediately with ample water as
first aid, and consult the doctor if required.
3) Methanol (HPLC grade) used for pretreatment
is highly flammable liquid, and is known forstrong eye irritancy, etc. When using Methanol,
please handle it very carefully following its
Material Safety Data Sheet.
2. Precautions for use
1) This product should be stored as directed,
without freezing. Freezing can deteriorate thereagents, which can produce inaccurate results.
Therefore, avoid using the reagents which havebeen previously frozen.
2) Do not use expired reagents. Use of suchreagents cannot guarantee the reliability ofmeasurement values.
3) Do not replenish the reagents.
4) After completion of measurement, this productshould be tightly stoppered and stored in arefrigerator.
5) Do not mix materials from different kit lot
numbers.
6) Do not perform the assay under direct sunlight
3. Precautions for Disposal
1) Before disposal, used samples and theircontainers must be immersed in sodium
hypochlorite solution at a concentration ofgreater than 0.1% for longer than 1 hour or
autoclaved at 121ºC for 20 minutes.
2）To prevent infections from spilled samples orsolutions containing samples, wipe the spilledarea thoroughly with disinfectants such as
sodium hypochlorite solution at a concentrationof greater than 0.1%.
3) The reagents and treated samples should bediscarded as medical waste or industrial waste
according to the waste disposal regulations.
4) The reagents should be disposed of inaccordance with the Water Pollution Control
act or related regulations.
5) Sodium azide has been added as an antiseptic
agent in the TACR Antibody Solution 1 andTACR Latex Reagent 2. It can react with leador copper pipes to produce the highly explosive
metal azide. Therefore, the reagent should beflushed with large amounts of water during
disposal.
4. Other precautions
Do not use the containers for other purposes.

Packaging
Name
TACR Antibody
Solution 1
Nanopia TDM TACR Latex
Tacrolimus
Reagent 2
Pretreatment
Solution 3

Package1 × 13 mL
1 × 8 mL
2 × 15 mL

References
1) Drug Interview Form for Prograf Capsules 0.5mg,
1mg, 5mg, Prograf Granules 0.2mg, 1mg.
Japanese.
2) SEKISUI MEDICAL CO., LTD. In house data.
Japanese.
Contact
SEKISUI MEDICAL CO., LTD.
international@sekisui.com

Manufacturer
SEKISUI MEDICAL CO., LTD.
1-3, Nihonbashi 2-chome, Chuo-ku, Tokyo, Japan"Nanopia" is trademark or registered trademarks of
SEKISUI MEDICAL CO., LTD. in Japan and/or
other countries.
All other brands, product names and service names
are trademarks or registered trademarks of theirrespective companies.

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