OratecPlus

Device for drug screening
and alcohol in the oral fluid
ME / TH / CO / AM / OP / PC or BZ and alcohol
Intended use

CE Medical Device.
OratectPlus for the screening of drugs and alcohol in the oral fluid is an in vitro diagnostic device exclusively for use by medical personnel. OratectPlus for oral drug and alcohol screening is intended for use in the simultaneous detection of various drugs or related metabolites and alcohol in the human oral fluid. The device provides qualitative results for cocaine, methamphetamine, MDMA (ecstasy), THC (cannabinoids), amphetamine, opiates, phencyclidine, benzodiazepines and alcohol in human oral fluid at the following cutoff concentrations:

MYSELF	d-Methamphetamine / MDMA	25 ng / ml
TH	Δ9-Tetrahydrocannabinol	40 ng / ml
CO	Cocaine	20 ng / ml
AM	d-Amphetamine	25 ng / ml
OP	Morphine	10 ng / ml
PC	phencyclidine	4 ng / ml
BZ	Diazepam	5 ng / ml
TO THE	Alcohol	≥ 0.02% BAC

The test must be performed by adequately trained personnel. It must not be used without supervision.

OratectPlus for drug and alcohol screening in the oral fluid provides only preliminary results for the alcohol / drug test. For qualitative results or for the confirmation of presumptive positive results obtained with the use of OratecPlus for loscreening of drugs and alcohol in the oral fluid, it is necessary to use a more specific alternative method.

Summary
The use of drugs and / or the consumption of excessive amounts of alcohol contribute to the occurrence of many accidents with consequent injuries and permanent porters. Individual screening for drug abuse and alcohol detection is an important method for identifying individuals who could potentially harm themselves and others.

Studies on methamphetamine, MDMA, cocaine, opiates, amphetamine, phencyclidine, benzodiazepine, cannabinoids and alcohol show that all these substances are detectable in oral fluid. OratectPlus for the screening of drugs and alcohol in the oral fluid integrates the collection of oral fluid and the simultaneous identification of drugs of abuse and alcohol in a single device.

Test principle
OratectPlus for oral drug and alcohol screening is based on:

(1) Drug test: a competitive lateral-flow enzyme immunoassay in which derivatives of immobilized membrane drugs compete with the drug (or drugs) that could be present in the oral fluid for a limited number of binding sites on the antibody-colloidal gold conjugate colored. During the test the oral fluid is deposited on the collection pad and from there migrates through the membrane. If no drugs are present in the fluid, the colored colloidal antibody-gold conjugate binds aiderivatives present on the membrane to form visible bands in specific areas of the test. Therefore, the presence of a colored band in a specific area of the test indicates a negative result . In the event that any of the drugs indicated above is present in the oral fluid, it competes with the immobilized conjugate for the limited number of binding sites of the colloidal conjugate with colored gold. A sufficient amount of drug is present in the sample, the antibodies break down and the colloidal conjugate with gold cannot bind to the derivatives of the membrane drug. Therefore, the absence of a colored band in the test area indicates a presumptive positive result for that particular drug .

Fig. A Description of the test areas of the OratectPlus device for the screening of oral and alcohol fluids.

The presence of a blue line in each window indicates that the device has not yet been used. Exceeding the blue line indicates that a sufficient amount of oral fluid has been collected.

A control band in the appropriate area (C) indicates that the test has been performed correctly. This control band must always appear, independent of the presence of drugs or their metabolites.

(2) Alcohol test: in contact with the alcohol present in the oral fluid, a buffer covered with enzymes changes color gradually passing through the variety of green and blue. The alcohol test buffer uses a solid phase agent that uses the following highly specific enzymatic reaction:

	Alcohol oxidase
CH 3 CH 2 OH + O 2	-------------->	CH 3 C = O + H 2 O 2
	peroxidase
H 2 O 2 + DH 2	-------------->	D Blue + 2H 2 O

During the test, the oral fluid is collected on the pad and saturated. If alcohol is not present in the fluid, the buffer remains colorless (it remains white or colorcrema) because no reaction takes place with enzymes that can cause discoloration. When alcohol is present in the oral fluid, the sicolora buffer of a shade between green and blue because alcohol reacts by producing aldehyde and peroxide. Hydrogen liberated through the peroxidase reaction which leads to the formation of the blue color. Therefore, the presence of a green-blue color on the pad indicates a presumed positive result for the alcohol test.

Reagents
OratectPlus for drug and alcohol screening in the oral fluid contains:

(1) Test for drugs: the drug test consists of two membrane strips and a collection pad. Each strip consists of a membrane, a colloidal gold conjugate pad, a swab for the sample and a swab absorbent.

Membrane : test strip for ME / TH / CO In a specific area of the membrane, referred to as the "Test Zone", dimethanfetamine, THC and cocaine protein conjugates were absorbed.

Test strip for AM / OP / PC or AM / OP / BZ: amphetamine, morphine, and phencyclidine, amphetamine, morphine and benzodiazepine conjugates have been absorbed into the membrane test area.

Conjugate gold buffer with colloidal gold : the buffer strip of the colloidal conate conjugate for the ME / TH / CO test contains conjugates with colloidal gold of methamphetamine antibodies, anti THC and anti cocaine absorbed on a fiber pad. The colloidal conjugate with colloidal gold for the AM / OP / PC or AM / OP / BZ test contains colloidal conoroidal conjugates of anti-amphetamine antibodies, anti morphine and anti phencyclidine or anti-amphetamine antibodies, anti morphine and anti benzodiazepine.

(2) Alcohol test - The alcohol pad contains
Tetramethylbenzidine Alcohol oxidase
peroxidase
Buffer e
Stabilizing proteins

Material supplied

OratectPlus for drug and alcohol screening in the oral fluid contains:
1. 1 insert
2. 1 guide
3. 25 test devices: each device consists of a plastic support and a removable cap. The devices are individually packaged in an aluminum bag with desiccant and oxygen absorbent material.
4. 1 plastic vial containing the confirmatory test swab

Materials requested but not supplied
• Time measurement device

Warnings and precautions
• OratecPlus for drug and alcohol screening in the oral fluid is indicated exclusively for in vitro diagnostic use.
• The test device must remain in the original packaging until it is used.
• If the package is torn or cut the device must be discarded.
• Do not use the device beyond the expiry date stated on the package.
• Treat all saliva samples as potentially infected. Proper procedures for the treatment and disposal of the product must be established.
• The results should not be interpreted by color-blind or disturbing people.

The alcohol test has been studied and calibrated to be interpreted 5 minutes after the start of the saliva collection procedure. If you wait for more than ten minutes, incorrect or false results may occur. The alcohol test is sensitive to the presence of ethanol. Sometimes the alcohol vapors in the air are detected by the OratecPlus device. Alcohol fumes are often present in public spaces and homes. Alcohol is a component of many household products, such as disinfectants, deodorisers and liquid for glass cleaning. If the presence of alcohol vapors is suspected, it is necessary to expose yourself and perform the test in an area that is certainly free of vapors.

The alcohol test is a visual interpretation in which each color change from green to blue is considered to be a presumed positive result with an alcohol concentration of 0.02% or higher in saliva.

Product storage
The unopened package of the OratecPlus device must be stored at room temperature, 15 ° -30 ° C (59 ° F-86 ° F), which must not exceed 30 ° C (86 ° F) . If stored in compliance with these conditions the alcohol test will follow the specifications up to the expiration date. If the storage temperature exceeds 30 ° C (86 ° F), degradation of the product and its performance may occur. Do not open the package until the test is completed.

Sample collection and processing
IMPORTANT: at least 10 minutes before the test is administered, ask the subject not to eat, drink, smoke or chew chewing gum or food based products because these behaviors can lead to incorrect results due to possible contamination of the saliva with interfering substances.

Test procedure
1. Remove the device from the sealed package.
2. Carefully remove the cap by holding it on the sides and pulling gently. In this way the collection pad and a circular pad are exposed for the alcohol content between the band and the pad.
3. Check that the blue line is present in each test band.
4. Observe the alcohol pad. The swab should be light cream in color. If the color is dark or in any way discolored before taking the saliva, the device should not be used.
5. The oral fluid collection procedure must be carefully observed. Indicate the subject to hold the device from the top (above the test window).
6. When inserting the device into the mouth, keep the head horizontal .
to. Open your mouth, insert the collection pad inside and gently roll it out several times against a cheek with a circular motion (about 15-20 times). (Fig. B)
b. Keeping the head horizontal, rub the collection pad against the opposite cheek several times (about 15-20) with a circular motion. (Fig B)

Fig. B Rub the collection pad against each end several times (about 15-20).

c. Rub the collection pad on the tongue several times (about 15-20) and then under the tongue several more times (about 15-20). (Fig. C and Fig. D). Do not chew, suck, bite or bend the collection pad.

Fig. C Rub the collection pad on the tongue several times (about 15-20).

Fig. D Rub the collection pad under the tongue several times (about 15-20).

7. Put the collection pad under the tongue for about 30 seconds to collect the laser. Instruct the donor to hold the device in place with his or her hands.
8. The flow of blue lines indicates the collection of a sufficient quantity of waste. If the blue line is still present after placing the collection pad under the tongue for 30 seconds, repeat the instructions from point 5 to point 7 until the blue line disappears (the absorption of the saliva exceeds the line)

Note: The flow of saliva over the blue line must occur in the two testentro bands five minutes. If no flow changes are observed after 5 minutes within the mouth, discard the device, review steps 4 to 7 of the procedure and repeat the test using a new device.

9. Put the cap back on the device, place it on a flat surface and read the results:
(a) The alcohol test must be read 5 minutes after removal of the device from the mouth. Do not read the result after 10 minutes. (b) The drug test should be read 5 minutes after removing the device from the mouth. Do not read the result after 30 minutes.

Interpretation of test results
(1) Alcohol test results:
(A) Negative result for alcohol When the alcohol pad shows no color changes (it remains white or cream), the result must be interpreted as negative (no alcohol is present). See Fig. E for an example of a negative alcohol test result.

In Fig. E below, the oral fluid sample is negative for alcohol because no color changes have occurred in the alcohol pad.

Fig E. Example of a negative result of the alcohol test

A result in which the outer edges of the alcohol pad stains weakly the majority of the pad remains colorless must be repeated to ensure complete saturation of the pad with oral fluid. If the second test is equal to the first, the result must be interpreted as negative (absence of alcohol).

(B) Presumptive positive result for alcohol: the alcohol test produces a color change towards green or blue in the presence of an alcohol concentration in the saliva equal to 0.02% BAC or higher. At higher dialcol concentrations, close to 0.30% BAC, the color can become gray-gray.

In Fig. F the oral fluid sample is positive for alcohol because there have been color changes (from green / blue to dark gray) of the alcohol pad.

Fig F. Example of a positive presumed result of the alcohol test

(2) Drug test results:

(A) Negative result for drugs : in each of the windows of the edema test, colored lines must be observed: one in the two control zones (C) and another next to the specific abbreviation of the drug (eg AM, OP, CO) in the test area (T). See the example in Fig. G.

The color of the test band may be slightly darker or brighter than the control band. Any band that can be visually noticed, no matter how weak, must be interpreted as a negative result. Read each test independently. Don't compare the intensity of different tests.

In Fig. G below, the sample of oral fluid is negative peranfetamine, opiates and cocaine because bands are visible in the zonedel tests related to AM, OP and CO .

Fig G. Example of a negative drug test result

(B) Presumptive positive result for drugs : when the corresponding band is visible in the control region (C) and no band appears in the specific region of the test, the result must be deemed presumptive positive for that particular drug. In Fig. H below, the oral fluid sample is presumptive positive for phencyclidine, methamphetamine and THC because there are no visible bands in the test areas related to AM, OP and CO .

Fig H. Example of a presumptive positive drug test result

(c) Invalid results : when no bands appear in the control area (C), the test should not be considered valid regardless of the result obtained in the test area. If the test is invalid, check the test procedures and samples. Repeat the test using a new device. In Fig. I below, the test is invalid because there are no bands in the control areas .

Fig. I Example of invalid test results

Important: read each test independently. Don't compare the intensity of different bands. If a faded band is obtained in the test area for a specific test, the sample should be considered negative. The OratectPlus device for drug and alcohol screening in the oral fluid provides qualitative results for the presence of drugs or drugs and alcohol at a specific cutoff concentration. For confirmation of a presumptive positive result with regard to drugs of abuse, a more specific quantitative method (GC / MS or LC / MS) should be used.

Collection and management of the sample for the confirmation test
• For devices with any expected positive results, the collection pad must be removed and sent to a laboratory for confirmation testing.
• Insert the transparent cap on the collection pad and detach both. Make sure not to damage or alter the collection pad.
• Place the collection pad in the confirmation vial included in the package.
• Recap the vial and send it to the laboratory for the confirmation test (the sample must be stored at 15-30 ° C and analyzed as soon as possible after collection).
• Follow standard procedures for chain of custody.
• To confirm a presumptive positive alcohol test, use a theatrometer or blood test.

Quality check
The OratectPlus device for the screening of drugs and alcohol in the oral liquid is equipped with integrated control bands positioned in the control zones (C) which serve to indicate that the test was performed correctly. These control bands must always appear, regardless of the presence of drugs. The presence of colored bands in the control zones verifies that,

1. a correct sample volume was used
2. you have managed to get the correct flow.

If the control bands do not appear, the device must be discarded.

Limitations of the procedure
• The test is intended for exclusive use with human oral fluid.
• Positive results only indicate the presumed presence of drugs and / or alcohol. However, it does not indicate or measure a state of intoxication.
• Technical and procedural errors, in addition to certain types of substances present in some products and medicines, can interfere with the test and give rise to false results.

Test performance
Precision - Drug abuse test
For each individual drug test, an artificial oral fluid solution was added to a standard drug in different concentrations (0%, 50%, 200% and 300%). For each concentration, a total of 20 tests were performed to evaluate leprestazioni. The table below summarizes the results obtained with the OratectPlus device for drug and alcohol screening in the oral fluid.

50% 300% + - + 0 0 20 0 0 20 0 1 20 0 0 20 0 0 20 0 0 20 0 0 20 0 1 20

Type of training	Total number of tests / Concentration	Concentration
		0%		200%
		-	-	+	+	-
MYSELF	20	20	20	0	20	0
MDMA	20	20	20	0	20	0
TH	20	20	20	0	19	0
CO	20	20	20	0	20	0
AM	20	20	20	0	20	0
OP	20	20	20	0	20	0
PC	20	20	20	0	20	0
BZO	20	20	20	0	19	0

Precision - Alcohol test
For the alcohol test, saliva was obtained by rinsing with positive control solutions in ethanol at various BACs (0.02%, 0.04% and 0.06%). A negative saliva sample was used to test the 0% concentration. For each concentration, a total of 15 tests were performed to evaluate the performance. The table below summarizes the results obtained with the OratectPlus device for drug and alcohol screening in the oral fluid:
Test BAC 12:04% - + - 15 14 0

Total number of tests / Concentration
	0.00%		12:02%		12:06%
	+	-	-	+	+
Alcohol	15	0	1	0	15	15

Specificity
Specificity tests were performed for each drug adding compostestructurally related to an artificial oral fluid solution. The results are expressed as a quantity of compound, in ng / ml, which produced a positive result.

Approximate% of
cross reactivity 2.5% 8.3% 100% 83% 100% 5% 1% 0.5% 5000 4000 80 40% 40 400% 10 10% 110% 100% 0.4% 100% 62.5% 3.2% 200 40 100 100 0.3% 0.3% 50% 83% 100% 100% 17% 67% 17% 14% 100% 25 100 4 10 12 3 30 8 20 6 5 3 10 10 100 15 20 50 15 15 10

Type of training	Concentration approximate (ng / ml)
ME / MDMA
Desipramine	10000	0,25%
d, l-ephedrine	1000	2.5%
1R, 2S l-Ephedrine	1000	2.5%
p-Idrossimetanfetamina	1000
MDEA	300
MDMA	25
d, l-methamphetamine	30
d-Methamphetamine	25
I-Methamphetamine	500
Metossifenamina	2500
Phenylephrine	5000
d-Pseudoephedrine HCl	0.5%
trimethobenzamide	0.6%
TH
cannabinol	50%
Δ-8-Tetrahydrocannabinol	100
Δ-9-Tetrahydrocannabinol	100%
11-nor-Δ-8-THC-9-COOH	10
11-nor-Δ-9-THC-9-COOH	400%
11-Hydroxy-Δ9-THC	400
CO
benzoylecgonine	18
Cocaine	20
ecgonina	5000
AM
d-Amphetamine	25
d, l-amphetamine	40
I-Amphetamine	800
d, Ip-Cloranfetamina	12.5%
MDA	62.5%
MDEA	25%
Phentermine	25%
β-Phenylethylamine	8000
tyramine	8000
OP
6-Acetilcodeina	20
6-acetylmorphine	12
Codeine	10
dihydrocodeine	10
Ethyl morphine	60
Heroin	15
Hydrocodone	60
Hydromorphone	70
Morphine	10
Morphine-3-beta-D-glucuronide	40%
nalorphine	10%
PC
phencyclidine	100%
BZ
Alprazolam	50%
bromazepam	42%
Chlordiazepoxide	167%
clobazam	16.7%
Clonazepam	63%
delorazepam	25%
Diazepam	83%
estazolam	100%
flunitrazepam	167%
flurazepam	50%
Lorazepam	50%
lormetazepam	5%
Nitrazepam	33%
nordiazepam	25%
oxazepam	10%
Prazepam	33%
Temazepam	33%
triazolam	50%

Alcohol test
The alcohol test reacts with methyl, ethyl and allyl alcohols, but does not react with alcohols with more than five carbon atoms such as glycerin, glycerol eserine. This property is the result of the specificity of the enzyme alcohol oxidase extracted from yeast.

Interference
The following compounds were added to an artificial summer fluid solution established that no cross-reactions occur with the OratectPlus device for drug and alcohol screening in the oral fluid at the concentration of 10 µg / ml (10,000 ng / ml):

Acetaminophen Lithium salt of acetacetic acid Acetone 6-Acetylcodeine (except OP test) 6-Acetylmorphine (except for OP tests) Acetylsalicylic acid Albumin Allobarbital Alphenal Alprazolam (except BZ analysis) Amitriptyline amobarbital amoxapine Amoxicillin d-Amphetamine (except AM test) d, l-Amphetamine (except AM test) l-Amphetamine (except AM test) Ampicillin Apomorphine Aprobarbital l-Ascorbic acid Aspartame Atropine barbital Benzyl acid benzocaine Benzoic acid Hydro benzoylecgonine (except CO test) Bilirubin Alprazolam (except BZ analysis) d-brompheniramine Buprenorphine Butalbital Butetal Caffeine cannabidiol Cannabinol (except TH test) chloral hydrate Chlordiazepoxide (except BZ analysis) Chloroanphetamine (DL-p-) Hydrochloride (except AM test) Chloroquine d-Chlorpheniramine chlorpromazine Cholesterol Clobazam (except BZ analysis) Clomipramina Clobazam (except BZ analysis) Cocaine (except CO tests) Codeine (except OP tests) Cortisone l-Cotinine Creatine Creatinine cyclobenzaprine Delorazepam (except BZ analysis) Desoxycortisone acetate Desipramine (except ME tests) dextromethorphan Diazepam (except BZ analysis) Dihydrocodeine (except OP tests) 4-Dimetilaminoantipirina Diphenhydramine Dopamine (3-Hydroxytyramine) Doxepin hydrochloride doxylamine Ecgonine (except CO tests) Ecgonine methyl ester d, E-Ephedrine (except AM test) l-Ephedrine 1R, 2S l-Ephedrine (except ME tests) 1S, 2R d-Ephedrine l-epinephrine Erythromycin Estazolam (except BZ analysis) b-Estradiol Estrone-3-sulfate potassium salt Ethanol Ethyl morphine (except OP tests) Ethylidin-1,5-dimethyl-1-3,3-diphenylpyrrolidine perchloric salt Estazolam (except BZ analysis) Flurazepam (except BZ analysis) Furosemide Gentisic acid Glucose glutethimide Guaiacol glyceryl ether Hemoglobin Heroin (except OP test) p-hydroxymethamphetamine (Pholderin) (except ME tests) Ibuprofen

Imipramine Hippuric acid Idroclorotizide Hydrocodone (except OP tests) hydrocortisone Hydromorphone (except OP test) 11-Hydroxy-Δ-9-Tetrahydrocannabinol (except TH test) d, I-Isoproterenol l-Isoproterenol HCl Lidocaine Lorazepam (except BZ analysis) Lormetazepam (except BZ analysis) MDA (except AM test) MDEA (except AM and ME tests) MDMA (except ME tests) meperidine d, l-methadone d-Methamphetamine (except ME tests) d, 1-Methamphetamine (except ME tests) l-Methamphetamine (except ME tests) Methoxyphenamine (except ME tests) 2-Methylamine-Propiophenone HCl methylphenidate Morphine (except OP tests) Morphine-3-beta-D-glucuronide (except OP tests) Nalidixic acid Nalorphine (except OP tests) Naloxone Naltrexone hydrochloride d-Naproxen Niacinamide Nitrazepam (except BZ analysis) Nordiazepam (except BZ analysis) Nordoxepin hydrochloride d, l-Norefedrina hydrochloride norethindrone d-norpropoxyphene Nortriptyline hydrochloride Oxalic acid Oxazepam (except BZ analysis) Oxolinic acid Oxycodone papaverine Penicillin-G (Benzilpenicillin) pentazocine Pentobarbital perphenazine Phenylcyclidine (except PC test) pheniramine Phenobarbital Phenothiazine Fentermine (except AM test) Phenylephrine (except ME tests) b-Phenylethylamine (except AM test) d, 1-Phenylpropanolamine hydrochloride Prazepam (except BZ analysis) Prednisolone procaine promazine Promethazine d-Propoxyphene protriptyline d-Pseudoephedrine HCl (except ME tests) quinidine Ranitidine Riboflavin Salicylic acid secobarbital Serotonin Sodium chloride sulfamethazine Sulindac Temazepam (except BZ analysis) Tetracycline -8-Tetrahydrocannabinol (except TH test) Δ-9-Tetrahydrocannabinol (trannetest TH) 11-nor-Δ-8-THC-9-COOH (except TH test) 11-nor-Δ-9-THC-9-COOH (except TH test) Thiamine thioridazine Triazolam (except BZ analysis) trifluoperazine Trimethobenzamide (except ME tests) Trimipramine Maleato tryptamine d, l-tryptophan Tiramina (except AM test) d, l-Tyrosine Uric acid Verapamil zomepirac

Alcohol test
The following substances may interfere with the OratecPlus device for the screening of drugs and alcohol in the oral fluid when using samples other than the fluid:

(1) Agents that promote color development: peroxides and strong oxidants
(2) Agents that inhibit color development:
reducing agents: ascorbic acid, tannic acid, pyrogallol, mercaptans and tosylates, oxalic acid and uric acid.
Bilirubin, L-dopa, L-methyldopa and metampirone.

The substances listed above do not normally appear in the oral fluid in sufficient quantity to interfere with the test. However, special care must be taken to prevent them from being introduced into the mouth during the 15 minutes preceding the test.

Bibliography of suggested readings
1. Wong, R. The Current Status of Drug Testing in the US Workforce, American Clinical Lab, vol. 21 (1), page 21-23, 2002.
2. Caplan, Y. and Goldberger, B., Alternative Specimens for Workplace Drug Testing, J.Analytical Toxicology, vol. 25, p. 396-399, 2001.
3. Schramm, W., Smith, R. and Craig, P., Drugs of Abuse in Saliva: A Review, J.Analytical Toxicology, vol. 16, p. 1-9, 1992.
4. Mandatory Guidelines for Federal Workplace Drug Testing Programs, April 13, 2004 (69FR 19644).
5. Wong, R. On-site Oral Fluid Drug Testing by Oratect, in Drugs of Abuse: Body Fluid Testing, Wong, R and Tse, H ed., Humana Press, p146-158, 2005.
6. Bergmeyer, HU, Grabl, M & Walter, H. in Methods of Enzymatic Analysis, 3rd ed.Vol.II, ed. by Bergmeyer, HU, Verlag Chemie, Weinheim, 1983, P.143.
7. Blanke, RV in Fundamentals of Clinical Chemistry, ed. by Tietz, NW, WB SaundersCo., Philadelphia, 1970, P. 114
8. McCall, KEL, Whiting, B., Moore, MR & Goldberg, A., CLIN. SCI., 56, 283-286,1979.
9. Jones, AW, CLIN.EXP.PHARMACOL.PHYSIOL. 6, 53-59, 1979.
10. Jones, AW, CLIN.CHEM. 25, 1394 - 1398, 1979.

Code HMA11 / HM12