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Accusign nicotine 5894-d.p65

the binding of the dye coated with cotinine conjugate to the antibodies on References
the membrane. This prevents the formation of a line on the membrane.
1. U.S. Department of Health and Human Services, Public Health Service, Therefore, a cotinine-positive urine sample will not generate a line at the Centers for Disease Control: Reducing the Health Consequences of Smoking, 25 Years of Progress, A Report of the Surgeon General, AccuSign® Nicotine
Test position (T) in the Result Window, indicating a positive result frompositive cotinine competition, while a negative urine sample will generate Rockville, MD. Office on Smoking and Health, 1989.
a line at the Test position in the Result Window, indicating a negative result 2. U.S. Department of Health and Human Services, Public Health Service, One-Step Nicotine Test
from an absence of competition with free cotinine.
The Health Consequences of Smoking: Cardiovascular Disease, A In addition to the Test Line that may appear at the Test position (T), a Report of the Surgeon General, Rockville, MD. Office on Smoking and Control Line is present at the Control position (C) to confirm the viability of the test. This Control Lne (validation line) should always appear if the 3. Bruckert E, Jacob N, Lamaire L, Truffert J, Percheron F, and de Gennes test is conducted properly. This works as a procedural control, confirming JL. Relationship between Smoking Status and Serum Lipid in a Simple One-Step Immunoassay for the Qualita-
that proper sample volume was used and the reagent system at the control Hyperlipidemic Population and Analysis of Possible Confounding tive Detection of Nicotine Metabolite in Urine
Line and the conjugate-color indicator worked. If insufficient sample Factors. Clin Chem 1992;38:1698-1705.
volume is used, there may not be a Control Line, indicating the test is invalid.
4. Pojer. R, Whitfield JB, Poulos V, Eckhard IF, Richmond R, Hensley WJ. Carboxyhaemoglobin, Cotinine and Thiocyanate Assay Com- PBM
pared for Distinguishing Smokers from Non-smokers. Clin Chem Materials Provided
5. Benowitz NL, Kuyt F, Jacob P, et al. Cotinine Dispostion and Effects.
The AccuSign® Nicotine test kit contains all the reagents necessary to
Clin Pharmacol Ther 34, 604-611 (1983).
AccuSign® Nicotine device. The test device contains a membrane
Intended Use
strip and a dye pad: The membrane strip is coated with monoclonal Symbols Key
anti-cotinine antibody and the dye pad contains dye coated with The AccuSign® Nicotine test is a simple, one-step, immuno-chromato-
graphic assay for the rapid, qualitative detection of cotinine, a major metabolite of nicotine, at the cutoff of 500 ng/mL in human urine.
AccuSign® Nicotine is used as an aid in the detection of cotinine after use
of tobacco products or other products containing nicotine. For in vitro Materials Needed but Not Provided
The AccuSign Nicotine test provides only a preliminary analytical result.
In Vitro Diagnostic Medical Device A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC/MS) is the preferred confirmatory method. Summary and Explanation
Smoking has been identified as a major risk factor for lung cancer and For in vitro diagnostic use only.
“Use By” date in year-month-day format cardiovascular disease.1,2 Self-reporting of smoking status is not reliable.3 Avoid cross contamination of urine samples by using a new urine The detection of cotinine, a major metabolite of nicotine, has become the specimen container and dropper for each urine sample.
preferred biomedical method of assessing the smoking status of individuals Urine specimens are potentially infectious. Proper handling and on account of its sensitivity and specificity.4 disposal methods should be established according to good laboratory Cotinine is present in blood, urine, and saliva of individuals who smoke or chew tobacco or who inhale tobacco smoke produced by others. As an The AccuSign® Nicotine device should remain in its original sealed
objective indicator of nicotine intake or confirmation of nonsmoker status, pouch until ready for use. Do not use the test if the pouch is damaged cotinine offers several advantages over other biochemical measures: It is a specific indicator of nicotine intake, its concentrations are not influenced Do not use the test kit after the expiration date.
by confounding factors such as diet or environment, its average biological half-life in blood is 19 hours, and its concentration within a given individual Storage and Stability
varies by only 15 to 20% over the course of a day.5 Cotinine assay is thusa superior objective measure of exposure to nicotine.
The AccuSign® Nicotine test kit should be stored at 2–30°C (35–86°F)
in the original sealed pouch. The expiration date given was established under Principle
The AccuSign® Nicotine test uses solid-phase chromatographic mem-
Specimen Collection and Preparation
brane immunoassay technology for a qualitative detection of a nicotinemetabolite, cotinine, in human urine. The test is based on the principle of Approximately 110 μL of urine sample is required for each test. Fresh urine the highly specific immunochemical reactions between antigens and anti- specimens do not require any special handling or pretreatment. Specimens bodies which are used for the analysis of specific substances in biological should be collected in a clean glass or plastic container. If testing will not fluids. The test relies on the competition to bind to the antibodies between be performed immediately, specimens should be refrigerated (2–8°C) or the cotinine conjugate and cotinine that may be present in the urine sample.
frozen. The stability of specimens in a refrigerator or a freezer is established In the test procedure, a sample of urine is placed in the Sample Well of the for up to 5 weeks. Specimens should be brought to room temperature before AccuSign® is a Registered Trademark of Princeton BioMeditech Corpora-
device and is allowed to migrate upward. If cotinine is present in the urine sample, it competes with the cotinine conjugate, which is bound to the dye, MT Promedt Consulting GmbH
Specimens containing a large amount of particulate matter may produce Princeton BioMeditech Corporation
for the limited antibodies immobilized on the membrane. If the cotinine level inconsistent test results. Such specimens should be clarified by centrifuging is above the cutoff level, cotinine will saturate the antibodies, thus inhibiting or allowing settling before testing.
Table 1. AccuSign® Nicotine test vs an EIA test
The reproducibility of the test results of the AccuSign® Nicotine test was
examined at three different sites using a total of 15 blind controls, consisting If the procedure was followed properly.
of 5 negative samples, 5 at half cutoff level (250 ng/mL cotinine), and 5 If no Control Line appears, the test is NOT VALID.
positive samples (1,000 ng/mL cotinine). The results obtained at these three Repeat the test using a new device, and follow the sites with these controls demonstrated 100% agreement with expected Specificity
The specificity of AccuSign Nicotine test was determined by adding the
compounds structurally related to nicotine to cotinine-negative urine
In a separate study, the accuracy of AccuSign® Nicotine test was
Cotinine ()
Cotinine (+)
evaluated in comparison to the cotinine value of the sample as determined specimens and testing with the AccuSign Nicotine test kit. The results
by the GC/MS. The samples contained cotinine from 0 ng/mL to 2155 ng/ are expressed in terms of the concentration required to produce a positiveresult (Table 4).
mL by GC/MS. The result is shown in Table 2.
Table 2. Correlation with GC/MS value
Table 4. Specificity
Cotinine ()
Negative () = Control Line and Test Line
Positive (+) = Control Line only; No Test Line Test Procedure
This test detects only the presence of cotinine in urine. A positive test result does not provide any indication of intoxication or urinary The test procedure consists of adding the urine sample to the Sample Well of the device and watching for the appearance of colored lines in the Result The test result read after 10 minutes may not be consistent with the original reading obtained within the 10 minute reading period. The test must be read within 10 minutes of sample application.
Test Protocol
Certain medications containing cotinine may produce a positive result 1. For each test, open one AccuSign® Nicotine pouch and
in any chemical or immunological assay.
Interfering Substances
label the Status device with the patient ID.
2. Holding the dropper vertically, dispense 3 full drops (110 User Quality Control
A: Negative (smaller than 25% below cutoff ) Endogenous Compounds
μL) of the urine sample into the Sample well (S).
Internal Control: Each AccuSign® test device has built-in controls. The
B: Near Cutoff Negative (between 25% below cutoff and cutoff) AccuSign Nicotine test showed no interference when the endogenous
compounds were added at the concentration given below (Table 5) to urine
3. Read the result after 5 minutes, but within 10 minutes of Control Line is an internal positive procedural control. A distinct reddish- C: Near Cutoff Positive (between cutoff and 25% above cutoff) purple Control Line should always appear at the C position, if the test samples which had +25% cutoff concentration of cotinine.
D: Positive (greater than 25% above cutoff) procedure is performed properly, an adequate sample volume is used, thesample and reagent are wicking on the membrane, and the test reagents at Table 5. Endogenous compounds
Interpretation of Results
the control line and the conjugate-color indicator are reactive. In addition, All false positives samples contained cotinine greater than 350 ng/mL and if the test has been performed correctly and the device is working properly, Negative: Two lines. The appearance of two reddish-purple lines – one
possibly due to the cross-reactivity of nicotine’s metabolites. Total the background in the Result Window will become clear and provide a at the Test position (T) and the other at the Control position (C) in the
distinct result. This may be considered an internal negative procedural Result Window – indicates a negative test result; i.e., no cotinine above the cutoff level has been detected. The color of the Test Line may be weaker Precision
or stronger than that of the Control Line. A negative test result does not If the Control Line does not appear at the Control position, the test is invalid and a new test should be performed. If the problem persists, contact PBM AccuSign Nicotine test was determined by carrying
indicate the absence of cotinine in the sample. It indicates only that the sample out the test with serially diluted standard drug solutions (cotinine). Three does not contain cotinine above the cutoff level in qualitative terms. people carried out the study. The prepared concentrations were equal to Positive: One line. The appearance of only one reddish-purple line at the
External Control: External controls may also be used to assure that the
50% below cutoff, 25% below cutoff, cutoff, 25% above cutoff, 50% above Control position (C) in the Result Window and no distinct line at the Test reagents are working properly and that the assay procedure is followed cutoff and 100% above cutoff. There were no significant differences Exogenous Compounds
position (T) indicates the test result is positive (i.e., the specimen contains correctly. It is recommended that a control be tested at regular intervals as between operators, between days or between lots. Table 3 shows the The following compounds show no cross-reactivity when tested with cotinine at a concentration above the cutoff level).
good laboratory testing procedure and to follow federal, state, and local precision data that were combined all operators’ tests.
guidelines concerning the running of external quality controls. For infor- AccuSign Nicotine at a concentration of 100 μg/mL (Table 6).
Invalid: A distinct colored line should always appear at the Control
mation on how to obtain controls, contact PBM’s Technical Services.
position (C). The test is invalid if no line forms in the Control position (C).
Table 3. Precision Data
Table 6. Non Cross-Reacting Compounds
Note: A very faint line in the Test position (T), visible in 10 minutes, indicates
Expected Values
that the amount of cotinine in the sample is near or below the cutoff level for AccuSign® Nicotine is a qualitative assay. The amount of nicotine or
cotinine (a nicotine metabolite) present in the urine cannot be estimated by Limitations
the assay. The assay results distinguish positive from negative samples.
Positive results indicate the samples contain cotinine above the cutoff The test is designed for use with human urine only.
concentration and the individual has been exposed to nicotine.
There is a possibility that factors such as technical or procedural errors, as well as other substances not listed in the compounds tested in the Performance Characteristics
urine sample, may interfere with the test and cause erroneous results.
Adulterants, such as bleach and/or alum, in urine specimens may The performance of AccuSign® Nicotine was compared with commer-
produce erroneous results. If adulteration is suspected, the test should cially available Cotinine EIA test. The complete agreement (>99%) was observed. Results are shown in Table 1.

Source: http://www.drugtests.com.au/docs/AccuSign%20Nicotine.pdf


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