1



1.0 Name of kit: Sensatest Alcohol Saliva Test

2.0 Indication: For detection of elevated alcohol level in human saliva.

3.0 Package Size: One test per kit.

4. Active Ingredients :

1. Alcohol Oxidase

2. Horseradish Peroxidase

3. Substrate tetramethylbenzidine (TMB)

4. Buffer with protein stabilizer

5. Assay Protocol :

1. Assay Procedure:

1. Remove the test device from its foil pouch by tearing along the notch. Use the device as soon as possible.

2. Saturate pad end of stick with saliva in clean tub.

3. After complete saturation of pad, remove stick from saliva and read at the end of 2 minutes.

4. Compare reactive pad to colour blocks.

2. Interpretation of Results:

1. Negative: No colour change on reactive pad.

2. Positive: Reactive pad changes colour in 2 minutes.

5. Cut Off Value: 0.02% EtOH

6. Specificity:

1. Cross Reactivity

Results as shown in Table 1.

Table 1 – List of tested cross reactivity

|Substance |Concentration (ng/ml) |

|Formaldehyde |1,000 |

|Acetone |25,000 |

7. Storage: 2 ~ 30oC

8. Expiration Dating: 12 months from date of manufacture

10.0 Method of Manufacture:

1. Preparation of Reagent Chemicals

1. A batch of reagent chemical containing active ingredients is prepared at a specific ratio and quality tested for the proper pH and concentration.

2. Preparation of Reagent Pad

1. Paper pad is immersed into the reagent chemical and allowed to soak up the liquid. The pad is then removed from the reagent chemical and air dried.

3. Test Device Assembly:

1. The reagent pad is applied to a plastic backing and cut into strips.

2. The strips are sealed in aluminum pouches or plastic cans with desiccant.

11.0 Clinical Trials:

To establish the sensitivity and specificity of Sensatest One-step alcohol test kit relative to other qualitative saliva alcohol tests, 750 clinic samples were studied. A commercially available qualitative test kit (Chematics, Alco screen test) was used to compare against Sensatest alcohol saliva test for relative sensitivity and specificity in 750 saliva samples. Only 3 samples were discordant and the agreement is 99.67%. The detailed results are shown in Table 2.

Table 2 - Comparison between Chematics’ and SENSATEST’s alcohol tests

| |Chematics, Alco screen test |Subtotal |

| |+ |- | |

|SENSATEST Alcohol |+ |260 |3 |263 |

|Saliva test | | | | |

| |- |3 |484 |487 |

|Subtotal |263 |487 |750 |

Clinical specificity = TN ÷ (TN + FP), TN: True Negative, FP: False Positive

= 487 ÷ (487 + 3) = 99.4%

Clinical sensitivity = TP ÷ (TP + FN), TP: True Positive, FN: False Negative

= 263 ÷ (263 + 3) = 98.9%

12.0 Stability Studies:

The Alcohol Saliva Tests were tested for stability under three different conditions; 37ºC for 60 days, Room temperature for 12 months, and 2ºC for 12 months. Three lots of the samples were tested under each condition in the desiccated container. The data collected indicated that the sensitivity and specificity of the samples remained the same. The shelf-life for this product is determined to be 12 months. The results are shown in Table 3, 4 and 5.

Table 3 - Stability test in 37oC

|Alcohol |Lot number |Month |

| | |1 |2 |

|0.00% |Alcohol991012 |- |- |

| |Alcohol991205 |- |- |

| |Alcohol000125 |- |- |

|0.02% |Alcohol991012 |+ |+ |

| |Alcohol991205 |+ |+ |

| |Alcohol000125 |+ |+ |

|0.05% |Alcohol991012 |+ |+ |

| |Alcohol991205 |+ |+ |

| |Alcohol000125 |+ |+ |

Table 4 - Stability in room temperature

|Alcohol |Lot number |Month |

| | |1 |

| | |

| | |

|Amphetamine |1,000 |

|D/L-Amphetamine |2,000 |

|(±)3,4Methylenedioxyamphetamine |2,500 |

|L-Amphetamine |30,000 |

|(+)Methamphetamine |>100 mg/ml |

|(±)3,4Methylenedioxymethamphetamine |>100 mg/ml |

1. Interfering Substance

The following compounds exhibited no interference when dissolved in normal human urine at 100μg/ml (Table 2).

Table 2 - Non-Cross Reacting Compounds (100μg/ml)

|Acetaminophen |4-Acetamidophenol |Acetylsalicylic acid |

|Amikacin |Amitriptyline |Amobarbital |

|Arterenol |Aspartame |Ascorbic acid |

|Atrophine |Caffeine |Camphor |

|Chloroquine |Chlopheniramine |Cortisone |

|Deoxyephedrine |Dextromethorphan |Digitoxin |

|Digoxin |Diphenhydramine |Ecgonine |

|Ecgonine methyl ester |Ephedrine |Epinephrine |

|Gentisic |Guaiacol glycer ester |Histamine |

|Hydrochlorothiazide |Homatrophine |Imipramine |

|Ibuprofen |Isoproterenol |Ketamine |

|Lidocaine |Meperidine |Methadone |

|Methaqualone |Methylphenidate |Neomycin |

|Niacinamide |Perphenazine |Penicillin G |

|Phenylethylamine-a |Phenylpropanolamine |Promethazine |

|Pseudoephedrine |Quinine antidine |Salicyclic acid |

|Tetracycline |Tetrahydrozoline |Theophyline |

|Thioridazine |Trifluoperazine |Tryptophan |

|Tyramine | | |

9. Storage: 2 ~ 30oC

10. Expiration Dating: Minimum of 12 months from date of manufacture

10.0 Method of Manufacture:

4. Nitrocellulose Membrane Manufacture:

1. The Amphetamine-BSA conjugate diluted in phosphate buffered saline is coated on the test region. Simultaneously, goat anti-mouse IgG polyclonal antibody, diluted in phosphate buffered saline, is coated on the control region

2. The coated membrane is dried for a minimum of 24 hours, and then sealed in an aluminum bag which contains silica gel desiccant.

5. Anti-Amphetamine Mab colloidal gold conjugate pad manufacture:

1. A buffer solution containing mouse Anti-Amphetamine monoclonal antibody/colloidal gold conjugate is coated onto non-woven cloth sheets.

2. The coated non-woven cloth is dried for a minimum of 24 hours, and then sealed in an aluminum bag which contains desiccant.

6. Test Device Assembly:

1. The coated membrane S1, the conjugate pad S4, and an absorbent pad is applied to an adhesive-coated backing.

2. A 2-part waterproof label is applied over the conjugate pad and the absorbent pad, and the assembled sheet of material is cut into strips. The test strips are then vacuum-dried for a minimum of 4 hours.

3. The assembled test strip is sealed in an aluminum pouch along with a desiccant packet.

11.0 Clinical Trials:

To establish the sensitivity and specificity of Sensatest One-Step Amphetamine test kit relative to other detection of qualitative urine amphetamine testing, 435 samples including 185 positives and 250 negatives were studied at three independent test sites using Sensatest One-Step Amphetamine test kit. All Positive and Negative samples were determined using a commercially available SureStep One-Step Amphetamine (Applied Biotech, Inc) test and confirmed using GC/MS. In total , 5 samples were discordant and the agreement is 97.8%. The detailed results are as shown in Table 3.

Table 3 - Comparison of One-step Amphetamine with SureStep product for 435 cases

| |Results of SureStep |Subtotal |

| |Positive |Negative | |

|Results of SENSATEST |Positive |180 |1 |181 |

|kits | | | | |

| |Negative |4 |250 |254 |

|Subtotal |184 |251 |435 |

11.1 Clinical Specificity:

Clinical Specificity is defined as the probability of a negative result in the absence of the particular condition. Clinical Specificity was determined by assaying blind-coded Amphetamine samples. We classified as negative a sample that when tested with the Sensatest One-step Amphetamine Test Region.

Clinical Specificity = TN ( (TN+FP), where TN: True Negative, and FP: False Positive

Clinical Specificity = 250 ( (250+4) = 98.4%

11.2 Clinical Sensitivity:

Clinical Sensitivity is defined as the probability of a positive result in the presence of the particular condition. Clinical Sensitivity was determined by assaying blind-coded Amphetamine positive samples. We classified as Positive a sample that when tested with the Sensatest One-step Amphetamine Test showed only one coloured back in the Control Region, with no coloured band in the Test Region.

Clinical sensitivity = TP ( (TP+FN), where TP: True Positive, FN: False Negative

Clinical Sensitivity = 181 ( (181+4) = 97.8%

12.0 Stability Studies:

It is well known that immunochemicals are quite stable at room temperature if the materials are kept in dry conditions. The chemicals used in the One-Step Amphetamine device are either dried on the membrane or chemical pad. The final test device is then sealed in an aluminum pouch with a desiccant. To establish the shelf-life and to predict the expiration date of the test, accelerated stability testing has been carried out in this study.

Three lots of One-Step Amphetamine test kit have been used for this study. The test devices in aluminum pouches were kept in the incubator at 37(C or room temperature at 4-30(C. The test kits were removed from their storage and tested with positive and negative control panels. The results are as shown in Table 4 and 5.

Table 4 - Results of Stability study on One-Step Amphetamine test kits at 37(C

|S(+)AMP |Lot # |Months |

|(ng/ml) | | |

| | |1 |

| | |

| | |

|Cocaine |300 |

|Benzoylecogonine |300 |

1. Interfering Substance

The following compounds exhibited no interference when dissolved in normal human urine at 100μg/ml (shown as Table 2)

Table 2 - Non-Cross Reacting Compounds (100μg/ml)

|Acetaminophen |Diazepam |Levorphanol |

|Acetone |4-Dimethylamino- |Lidocaine |

|Acetylsalicylic Acid |atipyrine |LSD |

|Albumin |Dopamine |Loperamide |

|Aminopyrine |Doxylamine |Lorazepam |

|Amitriptyline |Ecgonine |Maprotiline |

|D-Amphetamine |Ecgonine Methyl Ester |Meperidine |

|L-Amphetamine |(+/-)-Ephedrine |Methadone |

|Ampicillin |(+)-Ephedrine |Methamphetamine |

|Aspartame |(-)-Ephedrine |Methaqulone |

|Aspirin |(+)-Epinephrine |(+) 3,4-Methylenedioxy- |

|Atropine |Erythromycin |Amphetamine (MDA) |

|Sulfate Benzocaine |Estradiol |(+) 3,4-Methylenedioxy- |

|Benzoic acid |Ethanol |ethamphetamine(MDEA) |

|Bilirubin |Ethyl morphine |(+) 3,4-Methylenedioxy- |

|(+)-Brompheniramine |Furosemide |methamphetamine(MDMA) |

|Caffeine Ketamine |Glucose |Morphine |

|Cannabidiol |Graiacol glycery ether |Morphine-3-glucuronide |

|Chloroquine |Hemoglobin |Nalidixic acid |

|Chlorpromazine |Hydrocortisone |Naloxone |

|Clonazepam |Histamine |Naltrexone |

|Codeine |Hydrocodone |Naporphine |

|Cortisone |Hydromorphone |(+)-Naproxen |

|Creatinine |Imipramine |Norcodeine |

|Dexbrompheniramine |(+/-)-Isoproterenol |Tetracycline |

| | | |

|Dextromethorphan |Ketamine |-8-THC |

|(+/-)-Norephedrine |Procaine |-9-THC |

|(-)-Norpseudoephedrine |Promethazine |11-nor-9-THC-COOH |

|Nortriptyine |Propanol |Thebaine |

|Oxalic acid |d-Propoxyphene |Thioridazine |

|Oxpazem |d-pseduoephedrine |Trifluoperazine |

|Oxycodone |Quinidine |Trimipramine |

|Oxymorphone |Quinine |Tryptophom |

|PCP |Raboflavin |Tyramine |

|Penicillin-G |Ranitidine |Uric acid |

|Pentazocaine |Salicylic acid |Verapamil |

|Pentermine |Secobarbital |Vitamin C |

|Phencyclidine |Serotonin |Zomepirac |

|Phenobarbital |Sulfamethazine | |

|Phenothiazine |Sodium Chloride | |

|L-Phenylephrine |Sulindac | |

|β-Phenylethylamine |Temazepam | |

11. Storage: 2 ~ 30oC

12. Expiration Dating: Minimum of 18 months from date of manufacture

10.0 Method of Manufacture:

7. Nitrocellulose Membrane Manufacture:

1. The Benzoylecognine-BSA conjugate diluted in phosphate buffered saline is coated on the test region. Simultaneously, goat anti-mouse IgG polyclonal antibody, diluted in phosphate buffered saline, is coated on the control region

2. The coated membrane is dried for a minimum of 24 hours, and then sealed in an aluminum bag which contains silica gel desiccant.

8. Anti-Cocaine mAb colloidal gold conjugate pad manufacture:

1. A buffer solution containing mouse Anti-Cocaine monoclonal antibody/colloidal gold conjugate is coated onto non-woven cloth sheets.

2. The coated non-woven cloth is dried for minimum 24 hours, and then sealed in an aluminum bag which contains desiccant.

9. Test Device Assembly:

1. The coated membrane S1, the conjugate pad S4, and an absorbent pad is applied to an adhesive-coated backing.

2. A 2-part waterproof label is applied over the conjugate pad and the absorbent pad, the assembled sheet of material is die-cut, and the die-cut test strips are then vacuum-dried for a minimum of 4 hours.

3. The assembled test strip is sealed in an aluminum pouch along with a desiccant packet.

11.0 Clinical Trials:

To establish the sensitivity and specificity of Sensatest One-Step Cocaine test kit relative to other detection of qualitative urine cocaine testing, 720 samples including 241 positives and 479 negatives were studied at three independent test site using Sensatest One-Step Cocaine test kit. All Positive and Negative samples were determined using a commercially available SureStep One-Step Cocaine (Applied Biotech, Inc) test and confirmed using GC/MS. Totally 6 samples were discordant, the agreement is 99.1%. The detail results show as Table 3.

Table 3 - Comparison of One-step Cocaine with SureStep product for 720 cases

| |Results of SureStep |Subtotal |

| |Positive |Negative | |

|Results of SENSATEST |Positive |236 |4 |240 |

|kits | | | | |

| |Negative |2 |478 |480 |

|Subtotal |238 |482 |720 |

11.1 Clinical Specificity:

Clinical Specificity is defined as the probability of a negative result in the absence of the particular condition. Clinical Specificity was determined by assaying blind-coded Cocaine negative samples. We classified as negative a sample that when tested with the Sensatest One-step Cocaine Test Region.

Clinical Specificity = TN ( (TN+FP), where TN: True Negative, and FP: False Positive

Clinical Specificity = 478 ( (478+2) = 99.5%

11.2 Clinical Sensitivity:

Clinical Sensitivity is defined as the probability of a positive result in the presence of the particular condition. Clinical Sensitivity was determined by assaying blind-coded Cocaine positive samples. We classified as Positive a sample that when tested with the Sensatest One-step Cocaine Test showed only one coloured back in the Control Region, with no coloured band the Test Region.

Clinical sensitivity = TP ( (TP+FN), where TP: True Positive, FN: False Negative

Clinical Sensitivity = 240 ( (240+2) = 99.1%

12.0 Stability Studies:

It is well known that immunoreagents are quite stable at room temperature if the materials are kept in dry conditions. The chemicals used in the One-Step Cocaine device are either dried on the membrane or chemical pad. The final test device is then sealed in an aluminum pouch with a desiccant. To establish the shelf-life and to predict the expiration date of the test, accelerated stability testing has been carried out in this study.

Three lots of One-Step Cocaine test kit are used for this study. The test devices in aluminum pouches were kept in the incubator at 37(C or room temperature at 4-30(C. The test kits are then removed from their storage and tested with positive and negative control panels. The results are shown as Table 4 and 5.

Table 4 - Results of Stability study on One-Step Cocaine test kits at 37(C

|Cocaine |Lot # |Months |

|(ng/ml) | | |

| | |1 |

| | |

| | |

|11-nor-∆-9-Tetrahydrocannabinol |50 |

|11-nor--∆-8-Tetrahydrocannabinol |500 |

|∆-9-Tetrahydrocannabinol |20,000 |

|∆-8-Tetrahydrocannabinol |20,000 |

|Canabinol |50,000 |

|Cannabidiol |100,000 |

1. Intefering Substances

The following compounds exhibited no interference when dissolved in normal human urine at 100μg/ml (shown as Table 2)

Table 2 Non-Cross Reacting Compounds (100μg/ml)

|4-Acetaminophenol |Ecgonine Methyl Ester |Noscapine |

|Apomorphine |Ephedrine |d-Norpropoxyphene |

|Amobarbital |Epinephrine |Naloxone |

|Amitriptyline |Gentisic Acid |Neomycin |

|Aminopyrine |Glutethimide |Oxymetazoline |

|Amoxicillin |Hydrochlorothiazide |Phendimetrazine |

|D-Amphetamine |Hydralazine |Phenobarbital |

|L-Amphetamine |5-Hydroxytryptamine |l-Phenylephrine |

|Acetophenetidin |Ibuprofen |Phentermine |

|Benzoylecgonine |Iproniazid |Pentazocaine |

|Butabarbital Sodium |Isoproterenol |Prednisone |

|Benzphetamine |Imipramine |d-Propoxyphene |

|Chloral Hydrate |Ketamine |Promazine |

|Caffeine |Lidocaine |Quinidine |

|Chloroquine |Labetalol |Secobarbital |

|Chlorpheniramine |Meperidine |Sulindac |

|Cocaine |Methyprylon |Salicylic acid |

|Cannabidiol |Morphine sulfate |Thioridazine |

|Clonidine |Morphine |Tetrahydrocortisone |

|Diazepam |Methaqualone |Thiamine |

|Doxylamine |Methadone |D,l-Thyrixine |

|Dextromethorphan |3,4-methylenedioxy-amphetamine |Tyramine |

|Diclofenec |3,4-Methylenedioxy-methamphetamine |Tetrahydrozoline |

|Ecgonine Hydrochloride |Methylphenidate |Verapamil |

6. Storage: 2 ~ 30oC

7. Expiration Dating: Minimum of 12 months from date of manufacture

10.0 Method of Manufacture:

10. Nitrocellulose Membrane Manufacture:

1. The THC-BSA conjugate diluted in phosphate buffered saline is coated on the test region. Simultaneously, goat anti-mouse IgG polyclonal antibody, diluted in phosphate buffered saline, is coated on the control region

2. The coated membrane is dried for a minimum of 24 hours, and then sealed in an aluminum bag which contains silica gel desiccant.

11. Anti-THC mAb colloidal gold conjugate pad manufacture:

1. A buffer solution containing mouse Anti-THC monoclonal antibody/colloidal gold conjugate is coated onto non-woven cloth sheets.

2. The coated non-woven cloth is dried for minimum 24 hours, and then sealed in an aluminum bag which contains desiccant.

12. Test Device Assembly:

1. The coated membrane S1, the conjugate pad S4, and an absorbent pad is applied to an adhesive-coated backing.

2. A 2-part waterproof label is applied over the conjugate pad and the absorbent pad, and the assembled sheet of material is die-cut, and the die-cut test strips are then vacuum-dried for a minimum of 4 hours.

3. The assembled test strip is sealed in an aluminum pouch along with a desiccant packet.

11.0 Clinical Trials:

To establish the sensitivity and specificity of Sensatest One-Step Cannabinoids (THC) test kit relative to other detection of qualitative urine Cannabinoids testing, 435 samples including 185 positives and 250 negatives were studied at three independent test site using Sensatest One-Step Cannabinoids (THC) test kit. All Positive and Negative samples were determined using a commercially available SureStep One-Step Cannabinoids (THC) (Applied Biotech, Inc) test and confirmed using GC/MS. Totally 5 samples were discordant, the agreement is 97.8%. The detail results show as Table 3.

Table 3 - Comparison of One-step Cannabinoids with SureStep product for 435 cases

| |Results of SureStep |Subtotal |

| |Positive |Negative | |

|Results of SENSATEST |Positive |180 |1 |181 |

|kits | | | | |

| |Negative |4 |250 |254 |

|Subtotal |184 |251 |435 |

11.1Clinical Specificity:

Clinical Specificity is defined as the probability of a negative result in the absence of the particular condition. Clinical Specificity was determined by assaying blind-coded THC negative samples. We classified as negative a sample that when tested with the Sensatest One-step Cannabinoids Test Region.

Clinical Specificity = TN ( (TN+FP), where TN: True Negative, and FP: False Positive

Clinical Specificity = 250 ( (250+4) = 98.4%

11.2Clinical Sensitivity:

Clinical Sensitivity is defined as the probability of a positive result in the presence of the particular condition. Clinical Sensitivity was determined by assaying blind-coded THC positive samples. We classified as Positive a sample that when tested with the Sensatest One-step Cannabinoids Test showed only one coloured back in the Control Region, with no coloured band the Test Region.

Clinical sensitivity = TP ( (TP+FN), where TP: True Positive, FN: False Negative

Clinical Sensitivity = 181 ( (181+4) = 97.8%

12.0 Stability Studies:

It is well known that immunoreagents are quite stable at room temperature if the materials are kept in dry conditions. The chemicals used in the One-Step Cannabinoids (THC) device are either dried on the membrane or chemical pad. The final test device is then sealed in an aluminum pouch with a desiccant. To establish the shelf-life and to predict the expiration date of the test, accelerated stability testing has been carried out in this study.

Three lots of One-Step Cannabinoids (THC) test kit have been used for this study. The test devices in aluminum pouches were kept in the incubator at 37(C or room temperature at 4-30(C. The test kits were removed from their storage and tested with positive and negative control panels. The results are shown as Table 4 and 5.

Table 4 Results of Stability study on One-Step Cannabinoids (THC) test kits at 37(C

|THC-COOH (ng/ml) |Lot # |Months |

| | |1 |

| | |

|(+) Methamphetamine |1000 ng/mL |

|(±) 3,4MethylenedioxyMethamphetamine |1000 ng/mL |

|D-Amphetamine |> 100 |

|L-Amphetamine |> 100 |

|(±) 3,4Methylenedioxyamphetamine |> 100 |

|Chloroquine |> 100 |

|(-) Ephedrine |> 100 |

|ß-Phenylethylamine |> 100 |

|Procaine |> 100 |

|d-Pseudoephedrine |> 100 |

|Rantidine |> 100 |

1. Interfering Substance

The following compounds exhibited no interference when dissolved in normal human urine at 100 μg/ml as shown in Table 2.

Table 2 – Non-Cross Reacting Compounds (at 100 μg/ml)

|Acetaminophen |4-Acetamidophenol |Acetylsalicylic acid |

|Amikacin |Amitriptyline |Amobarbital |

|Arterenol |Aspartame |Ascorbic acid |

|Atrophine |Caffeine |Camphor |

|Chloroquine |Chlopheniramine |Cortisone |

|Deoxyephedrine |Dextromethorphan |Diphenhydramine |

|Digoxin |Ecgonine |Ecgonine methyl ester |

|Ephedrine |Epinephrine |Gentisic |

|Guaiacol glycer ester |Histamine |Hydrochlorothiazide |

|Homatrophine |Imipramine |Ibuprofen |

|Isoproterenol |Ketamine |Lidocaine |

|Meperidine |Methadone |Methaqualone |

|Methylphenidate |Neomycin |Niacinamide |

|Perphenazine |Penicillin G |Phenylethylamine-a |

|Phenylpropanolamine |Promethazine |Pseudoephedrine |

|Quinine antidine |Salicyclic acid |Tetracycline |

|Tetrahydrozoline |Theophyline |Thioridazine |

|Trifluoperazine |Tryptophan |Tyramine |

13. Storage: 2 ~ 30oC

14. Expiration Dating: Minimum of 12 months from date of manufacture

10.0 Method of Manufacture:

13. Nitrocellulose Membrane Manufacture:

1. The Methamphetamine-BSA conjugate diluted in phosphate buffered saline is coated on the test region. Simultaneously, goat anti-mouse IgG polyclonal antibody, diluted in phosphate buffered saline, is coated on the control region

2. The coated membrane is dried for a minimum of 24 hours, and then sealed in an aluminum bag which contains silica gel desiccant.

14. Anti-Methamphetamine mAb colloidal gold conjugate pad manufacture:

1. A buffer solution containing mouse Anti-Methamphetamine monoclonal antibody/colloidal gold conjugate is coated onto non-woven cloth sheets.

2. The coated non-woven cloth is dried for a minimum of 24 hours, and then sealed in an aluminum bag which contains desiccant.

15. Test Device Assembly:

1. The coated membrane S1, the conjugate pad S4, and an absorbent pad is applied to an adhesive-coated backing.

2. A 2-part waterproof label is applied over the conjugate pad and the absorbent pad, and the assembled sheet of material is cut into strips. The test strips are then vacuum-dried for a minimum of 4 hours.

3. The assembled test strip is placed into a plastic cassette casing.

4. The assembled cassette is sealed in an aluminum pouch along with a desiccant packet and disposable plastic pipette.

11.0 Clinical Trials:

To establish the sensitivity and specificity of Sensatest One-Step Methamphetamine test kit relative to other detection of qualitative urine Methamphetamine testing, 780 samples including 272 positives and 508 negatives were studied at three independent test site using Sensatest One-Step Methamphetamine test kit. All Positive and Negative samples were determined using a commercially available SureStep One-Step Methamphetamine (Applied Biotech, Inc) test and confirmed using GC/MS. In total, 8 samples were discordant and the agreement is 99.2%. The detail results show as Table 3.

Table 3 – Comparison of One-step Methamphetamine with SureStep product for 780 cases

| |Results of SureStep |Subtotal |

| |+ |- | |

|Results of SENSATEST kits |+ |268 |6 |274 |

| |- |2 |504 |506 |

|Subtotal |270 |510 |780 |

11.1 Clinical Specificity:

Clinical Specificity is defined as the probability of a negative result in the absence of the particular condition. Clinical Specificity was determined by assaying blind-coded Methamphetamine negative samples. We classified as negative a sample that when tested with the Sensatest One-step Methamphetamine Test Region.

Clinical Specificity = TN ( (TN+FP), where TN: True Negative, and FP: False Positive

Clinical Specificity = 504 ( (504+2) = 99.6%

11.2 Clinical Sensitivity:

Clinical Sensitivity is defined as the probability of a positive result in the presence of the particular condition. Clinical Sensitivity was determined by assaying blind-coded Methamphetamine positive samples. We classified as Positive a sample that when tested with the Sensatest One-step Methamphetamine Test showed only one coloured back in the Control Region, with no coloured band the Test Region.

Clinical sensitivity = TP ( (TP+FN), where TP: True Positive, FN: False Negative

Clinical Sensitivity = 274( (274+2) = 99.2%

12.0 Stability Studies:

It is well known that immunochemicals are quite stable at room temperature if the materials are kept in dry conditions. The chemicals used in the One-Step Methamphetamine device are either dried on the membrane or chemical pad. The final test device is then sealed in an aluminum pouch with a desiccant. To establish the shelf-life and to predict the expiration date of the test, accelerated stability testing has been carried out in this study.

Three lots of One-Step Methamphetamine test kit have been used for this study. The test devices in aluminum pouches were kept in the incubator at 42(C or room temperature at 2-30(C. The test kits were removed from their storage and tested with positive and negative control panels. The results are shown ad Table 4 and 5.

Table 4 – Results of Stability study on One-Step Methamphetamine test kits at 42(C

|S(+) |Lot # |Months |

|Methamphetamine (ng/mL) | | |

| | |1 |

| | |

| | |

|Morphine |300 |

|Codeine |300 |

|Ethyl morphine |300 |

|Hydrocodone |400 |

|Hydromorphone |400 |

|Levophanol |5,000 |

|Meperidine |80,000 |

|Morphine-3-glucuronide |500 |

|Naporphine |2,000 |

|Norcodeine |30,000 |

|Oxycodone |15,000 |

|Oxymorphone |50,000 |

|Thebaine |30,000 |

1. Interfering Substance :

The following compounds exhibited no interference when dissolved in normal human urine at 100μg/ml.

Table 2 - Non-Cross Reaction Compounds (100μg/ml)

|Acetaminophen |(+)-Brompheniramine |Ecgonine |

|Acetone |Caffeine |Ecgonine Methyl Ester |

|Acetylsalicylic Acid |Cannabidiol |(+/-)-Ephedrine |

|Albumin |Chloroquine |(+)-Ephedrine |

|Aminopyrine |Chlorpromazine |(-)-Ephedrine |

|Amitriptyline |Clonazepam |(+)-Epinephrine |

|D-Amphetamine |Cocaine |Erythromycin |

|L-Amphetamine |Cortisone |Estradiol |

|Ampicillin |Creatinine |Ethanol |

|Aspartame |Dexbrompheniramine |Furosemide |

|Aspirin |Dextromethorphan |Glucose |

|Atropine |Diazepam |Graiacol glycery ether |

|Sulfate Benzocaine |4-Dimethylamino- |Hemoglobin |

|Benzoic acid |Atipyrine |Hydrocortisone |

|Benzoylecogonine |Dopamine |Histamine |

|Bilirubin |Doxylamine |Imipramine |

|(+/-)-Isoproterenol |Oxalic acid |Secobarbital |

|Ketamine |Oxpazem |Serotonin |

|Levorphanol |Penicillin-G |Sulfamethazine |

|Lidocaine |Pentazocaine |Sodium Chloride |

|Loperamide |Pentermine |Sulindac |

|Maprotiline |Phencyclidine |Temazepam |

|Methadone |Phenobarbital |Tetracycline |

|Methamphetamine |Phenothiazine |∆-8-THC |

|Methaqulone |L-Phenylephrine |∆-9-THC |

|(+) 3,4-Methylenedioxy- |β-Phenylethylamine |11-nor-∆-9-THC-COOH |

|amphetamine |Procaine |Thioridazine |

|(+) 3,4-Methylenedioxy- |Promethazine |Trifluoperazine |

|methamphetamine |Propanol |Trimipramine |

|Nalidixic acid |d-Propoxyphene |Tryptophom |

|Naloxone |d-pseduoephedrine |Tyramine |

|Naltrexone |Quinidine |Uric acid |

|(+)-Naproxen |Quinine |Verapamil |

|(+/-)-Norephedrine |Raboflavin |Vitamin C |

|(-)-Norpseudoephedrine |Ranitidine |Zomepirac |

|Nortriptyine |Salicylic acid | |

8. Storage: 2 ~ 30oC

9. Expiration Dating: Minimum of 18 months from date of manufacture

10.0 Method of Manufacture:

16. Nitrocellulose Membrane Manufacture:

1. The Morphine-BDA conjugate diluted in phosphate buffered saline is coated on the test region. Simultaneously, goat anti-mouse IgG polyclonal antibody, diluted in phosphate buffered saline, is coated on the control region

2. The coated membrane is dried for a minimum of 24 hours, and then sealed in an aluminum bag which contains silica gel desiccant.

17. Anti-Morphine colloidal gold conjugate pad manufacture:

1. A buffer solution containing mouse Anti-Morphine monoclonal antibody/colloidal gold conjugate is coated onto non-woven cloth sheets.

2. The coated non-woven cloth is dried for minimum 24 hours, and then sealed in an aluminum bag which contains desiccant.

18. Test Device Assembly:

1. The coated membrane S1, the conjugate pad S4, and an absorbent pad are applied to an adhesive-coated backing.

2. A 2-part waterproof label is applied over the conjugate pad and the absorbent pad, and the assembled sheet of material is cut into strips. The test strips are then vacuum-dried for a minimum of 4 hours.

3. The assembled test strip is sealed in an aluminum pouch along with a desiccant packet.

11.0 Clinical Trials:

To establish the sensitivity and specificity of Sensatest One-Step Morphine test kit relative to other detection of qualitative urine cocaine testing, 780 samples including 272 positives and 508 negatives were studied at three independent test sites using Sensatest One-Step Morphine test kit. All Positive and Negative samples were determined using a commercially available SureStep One-Step Morphine (Applied Biotech, Inc) test and confirmed using GC/MS. A total of 8 samples were discordant and the agreement is 99.2%. The detailed results are shown as Table 3.

Table 3 - Comparison of One-step Morphine with SureStep product for 780 cases

| |Results of SureStep |Subtotal |

| |Positive |Negative | |

|Results of SENSATEST |Positive |268 |6 |274 |

|kits | | | | |

| |Negative |2 |504 |506 |

|Subtotal |270 |510 |780 |

11.1Clinical Specificity:

Clinical Specificity is defined as the probability of a negative result in the absence of the particular condition. Clinical Specificity was determined by assaying blind-coded Morphine negative samples. We classified as negative a sample that when tested with the Sensatest One-step Morphine Test Region.

Clinical Specificity = TN ( (TN+FP), where TN: True Negative, and FP: False Positive

Clinical Specificity = 504 ( (504+2) = 99.6%

11.2Clinical Sensitivity:

Clinical Sensitivity is defined as the probability of a positive result in the presence of the particular condition. Clinical Sensitivity was determined by assaying blind-coded Morphine positive samples. We classified as Positive a sample that when tested with the Sensatest One-step Morphine Test showed only one coloured back in the Control Region, with no coloured band the Test Region.

Clinical sensitivity = TP ( (TP+FN), where TP: True Positive, FN: False Negative

Clinical Sensitivity = 274( (274+2) = 99.2%

12.0 Stability Studies:

It is well known that immunoreagents are quite stable at room temperature if the materials are kept in dry conditions. The chemicals used in the One-Step Morphine device are either dried on the membrane or chemical pad. The final test device is then sealed in an aluminum pouch with a desiccant. To establish the shelf-life and to predict the expiration date of the test, accelerated stability testing have been carried out in this study.

Three lots of One-Step Morphine test kit have been used for this study. The test devices in aluminum pouches were kept in the incubator at 40(C or room temperature at 4-30(C. The test kits were removed from their storage and tested with positive and negative control panels. The results are shown ad Table 4 and 5.

Table 4 - Results of Stability study on One-Step Morphine test kits at 40(C

|Morphine |Lot # |Months |

|(ng/mL) | | |

| | |1 |

| | |

|Amphetamine | |

|d-Amphetamine |1,000 |

|l-Amphetamine |10,000 |

|(+/-)3,4-methylenedioxy- amphetamine |5,000 |

|phentemine |1,000 |

|Tenocyclidine |75,000 |

| | |

|Morphine | |

|Morphine |2000 |

|Codeine |2000 |

|Ethyl Morphine |2000 |

|Hydrocodone |25,000 |

|Hydromorphone |25,000 |

|Morphine-3-(--d-glucuronide |5000 |

|Thebaine |75,000 |

| | |

|Methamphetamine | |

|(+/-)3,4-Methamphetamine |1000 |

|D-Amphetamine |50,000 |

|Chloroquine |50,000 |

|(+/-)-Ephedrine |50,000 |

|(-)-Methamphetamine |25,000 |

|(+/-)3,4-methylenedioxymethampheta- |2000 |

|mphetamine(MDMA) | |

|(-Phemylethylamine |50,000 |

|Ranitidine |50,000 |

|Trimethobenzamide |10,000 |

| | |

|Cocaine | |

|Benzoylecogonine |300 |

|Cocaine |300 |

| | |

|THC | |

|11-nor-(9-THC-9-COOH |50 |

|11-nor-(8-THC-9-COOH |50 |

|11-hydroxy-(9-Tetrahydrocannabinol |2,500 |

|(8-Tetrahydrocannabinol |7,500 |

|(9-Tetrahydrocannabinol |10,000 |

|Cannabinol |10,000 |

|Cannabidiol |100,000 |

................
................

In order to avoid copyright disputes, this page is only a partial summary.

Google Online Preview   Download