1.0 Name of kit: Sensatest One-Step FSH Test – Cassette
2.0 Indication: For In Vitro Detection of Menopause - The FSH test is a qualitative immunoassay that detects elevated levels of Follicular Stimulating Hormone (FSH) in urine samples.
3.0 Package Size: One test per kit
4.0 Active Ingredients: 4.1 Coated Antibodies: 4.1.1 Control region: Goat anti-mouse (IgG) polyclonal antibody 4.1.2 Test region: Mouse monoclonal anti-FSH antibody A 4.2 Labeled Antibodies: 4.2.1 Colloidal gold conjugate of monoclonal anti-FSH antibody B
5.0 Assay Protocol: 5.1 Assay Procedure: Read the package insert for Sensatest One-Step FSH test carefully before starting the assay. Allow urine samples to warm up to room temperature before testing. 5.1.1 Remove the test device from its foil pouch, place it on a flat surface and label it with patient ID. Use the device as soon as possible. 5.1.2 Using a pipette, dispense 3-4 drops (0.2 ml) of urine onto the sample well of the cassette. 5.1.3 Wait 10 minutes and read results. It is important that the background is clear before the result is read. Results obtained after 30 minutes should be considered invalid.
5.2 Interpretation of Results: 5.2.1 Negative: Only one colour band appears on the control region or the test band is lighter than the control band. The FSH concentration of the sample is below 25 mIU/ml. 5.2.2 Positive: Two colour bands are visible and the test band is equal to or darker than the control band. The FSH is present in the sample at or above 25 mIU/ml. 5.2.3 Invalid: No visible band at all or no coloured line appears on the control region. Repeat test with a new test kit.
6.0 Cut off Value: 25 mIU/ml FSH in urine sample.
7.0 Specificity: 7.1 Cross Reactivity The cross reactivity of FSH test kits was evaluated with FSH homologous hormone. Homologous hormones, HCG, TSH, LH were added to urine samples containing FSH at concentration of 0, 25 or 100 mIU/ml. No cross reactivity was observed in the study (see Table 1).
Table 1 – Cross-reactivity study of One-step FSH test kit Urine samples spiked with homologous FSH conc. in Un-spiked hormones urine sample urine LH HCG TSH (mIU/ml) samples 1000mIU/ 1000mIU/ 1000mIU/ ml ml ml - - - - 0 ------+ + + + 25 + + + + + + + + + + + + 100 + + + + + + + +
7.2 Non-specific interference One-Step FSH test was checked for possible interference from visibly hemolyzed, lipemic and icteric samples. Human hemoglobin, bilirubin or albumin was spiked into samples with different concentration of FSH and tested using un-spiked sample as controls. No significant interference was observed in 20 sample testing results that were either positive or negative for FSH. The results are shown in Table 2.
Table 2 – Non-specificity study of One-step FSH test kits Urine samples spiked with (mg/ml) Un-spiked Sample No Hemoglobin Bilirubin Albumin samples 10 1 0.06 100 1 - - - - - 2 - - - - - 3 - - - - - 4 - - - - - 5 - - - - - 6 - - - - - 7 - - - - - 8 - - - - - 9 - - - - - 10 - - - - - 11 + + + + + 12 + + + + + 13 + + + + + 14 + + + + + 15 + + + + + 16 + + + + + 17 + + + + + 18 + + + + + 19 + + + + + 20 + + + + + 8.0 Storage: 2 to 30oC
9.0 Expiration Dating: 12 months from date of manufacture
10.0 Method of Manufacture: 10.1 Nitrocellulose Membrane Manufacture: 10.1.1 The purified monoclonal antibody, diluted in phosphate buffer saline, is coated on the test region. Simultaneously, goat anti-mouse IgG polyclonal antibody, diluted in phosphate buffer saline, is coated on the control region. 10.1.2 The coated membrane is dried for a minimum of 24 hours then sealed in an aluminum bag which contains silica gel desiccant. 10.2 Anti-FSH colloidal gold conjugate pad manufacture: 10.2.1 A buffer solution containing mouse Anti-FSH monoclonal antibody/colloidal gold conjugate is coated onto non-woven cloth sheets. 10.2.2 The coated non-woven cloth is dried for minimum 24 hours then sealed in an aluminum bag which contains desiccant. 10.3 Test Device Assembly: 10.3.1 The coated membrane S1, the conjugate pad S4, and an absorbent pad is applied to an adhesive-coated backing. 10.3.2 A 2-part waterproof label is applied over the conjugate pad and the absorbent pad, the assembled sheet of material is cut into strips. The test strips are then vacuum-dried for a minimum of 4 hours. 10.3.3 The assembled test strip is sealed in a plastic cassette casing. The cassette is sealed in an aluminum pouch along with a desiccant packet and disposable plastic pipette. 10.4 Packaging available :
Product Code Test Medium Test Format Packing Store
FSH 141 Urine Cassette 1 x 50 11.0 Comparison Study:
To establish the sensitivity and specificity of Sensatest One-step FSH test kits relative to other rates of qualitative urine FSH tests, 566 clinic samples were studied. Another commercially available qualitative test kit (Sytron FSH) was used to compare with Sensatest One-Step FSH test kit for relative sensitivity and specificity in 566 urine samples. In total, 3 samples were discordant. In turn, the agreement is 99.47%. The results are shown in Table 3.
Table 3 - Comparison of One-Step FSH with Sytron product for 566 cases
Results of SENSATEST kits Subtotal + - Results of Sytron + 278 1 279 kits - 2 285 287 Subtotal 280 286 566
12.0 Stability Studies: It is well known that immunochemicals are quite stable at room temperature if the materials are kept in dry condition. The chemicals used in the One-Step FSH device are either dried on the membrane or chemical pad. The final test device is then sealed in an aluminum pouch with a desiccant. Accelerated testing has been carried out in this study to establish the shelf life and to predict the expiration date of the test. Three lots of One-Step FSH test kit have been used for this study. The test devices in aluminum pouches were kept in the incubator at 42C or room temperature. Test kits were then removed from their storage and tested with positive and negative control panels. The results are shown in Table 4 and 5.
Table 4 - Results of Stability study on One-Step FSH test kits at 42°C FSH conc. 1 2 3 4 6 8 10 12 Lot # (mIU/ml) mo. mo. mo. mo. mo. mo. mo. mo. I04061C5A ------0 I04061C5B ------I04061C5C ------I04061C5A + + + + + + + + 25 I04061C5B + + + + + + + + I04061C5C + + + + + + + + I04061C5A + + + + + + + + 100 I04061C5B + + + + + + + + I04061C5C + + + + + + + + Table 5 - Results of Stability study on One-Step FSH test kits at room temperature FSH conc. 1 2 4 6 8 10 12 Lot # (mIU/ml) mo. mo. mo. mo. mo. mo. mo. I04061C5A ------0 I04061C5B ------I04061C5C ------I04061C5A + + + + + + + 25 I04061C5B + + + + + + + I04061C5C + + + + + + + I04061C5A + + + + + + + 100 I04061C5B + + + + + + + I04061C5C + + + + + + + +: FSH ≥ 25 mIU/ml -: FSH < 25 mIU/ml
The data collected to date indicate that One-Step FSH test kits are stable for at least 12 months at 2-30C.
13.0 Quality Control: 13.1 Appearance: All materials are visually inspected before their use in manufacture. 13.2 Specificity: 10 test kits are randomly selected and tested with Negative control panel (FSH < 25 mIU/ml). All test results must be negative. 13.3 Cut Off: 10 test kits are randomly selected and tested with a control panel in which contain 25 mIU/ml FSH. All test results must be positive. 13.4 Internal Control: A coloured band must appear in the control region of the membrane with each tested kit, which indicates proper performance and reagent reactivity. 13.5 Reproducibility: Randomly selected test kits from different lots must give the same results when assaying the same sample.
14.0 References: 14.1 Batzer FR, Fertility & Sterility, Vol 34, 1, 1980 14.2 Highton,J.and Hussian,P.,J.Immunol.Meth., 68:185,1984. 14.3 Braunstein GD, Rason J, Alder D, Danzer H, Wade ME, Am. J., Obstet. Gynecol., Vol 126, 677, 1976 14.4 Lenton E, Meal L, Sulaiman R, Fertility & Sterility, Vol 37, 773, 1982 14.5 Dawood M, Saeba B & Landesman R, Ob. Gyn., Vol 126, 678, 1976 14.6 Braunstein GD et al, Ann. Inter. Med., Vol 78, pp 39-45 14.7 Marshall,J.C.,Clinics in Endocrinol.Metab.,4:545,1975. 1.0 Name of kit: Sensatest One-Step HCG Test - Cassette
2.0 Indication: For In Vitro Detection of Early Pregnancy (HCG) - The one step HCG Urine Pregnancy Test is a combination of qualitative immunoassays for the determination of Human Chorionic Gonadotropin (HCG) hormone in urine samples. 3.0 Package Size: One test per kit
4.0 Active Ingredients: 4.1 Coated Antibodies:
4.1.1 Control region: Goat anti-mouse(IgG) polyclonal antibody 4.1.2 Test region: Mouse monoclonal anti-HCG antibody A
4.2 Labeled Antibodies:
4.2.1 Colloidal gold conjugate of monoclonal anti-HCG antibody B
6.0 Assay Protocol: 6.1 Assay Procedure: Read the test instructions for Sensatest One-Step HCG test carefully before starting the assay. Allow urine samples to warm up to room temperature before testing.
6.1.1 Remove the test device from its foil pouch, place it on a flat surface, and label it with the patient ID. Use the device as soon as possible. 6.1.2 Using a pipette, dispense 3-4 drops (0.2 ml) of urine onto sample well of the cassette. 6.1.3 Wait for the coloured bands to appear. Depending on the concentration of HCG in the test specimen, positive results may be observed in as short as 40 seconds. However, to confirm negative results, the complete reaction time (5 minutes) is required. It is important that the background is clear before the result is read. Do not read results after 10 minutes.
6.2 Interpretation of Results:
6.2.1 Negative (-): Only one pink or red coloured band appears on the control region(C). 6.2.2 Positive (+): In addition to the control band, a pink to red coloured band also appears on the test region (T). 6.2.3 Invalid: When no coloured band appears on the control region, the test should be repeated with a new test kit.
7.0 Sensitivity: 20 mIU/ml HCG in urine sample.
8.0 Specificity: 8.1 Cross Reactivity
The cross reactivity of HCG test kits was evaluated with HCG homologous hormones. Homologous hormones FSH, LH and TSH were added to urine samples containing HCG at concentration of 0, 20 or 100 mIU/ml. No cross reactivity was observed in the study as shown below in Table 1.
Table 1 - Cross-reactivity study of One-Step HCG test kit
HCG conc. Un-spiked serum Urine samples spiked with homologous hormones in sample or urine samples (mIU/ml) FSH LH TSH 1000 mIU/ml 1000 mIU/ml 1000 mIU/ml - - - - 0 ------+ + + + 20 + + + + + + + + + + + + 100 + + + + + + + +
8.2 Non-specific interference
One-Step HCG test was checked for possible interference from visibly hemolyzed, lipemic and icteric samples. Human hemoglobin, bilirubin or albumin was spiked into samples with different concentration of HCG and tested using un-spiked samples as controls. No significant interference was observed in 20 samples with results that were either positive or negative for HCG. The results are shown in Table 2.
Table 2 - Non-specificity study of One-Step HCG test kits Urine samples spiked with (mg/ml) Un-spiked Sample No Hemoglobin Bilirubin Albumin samples 10 1 0.06 100 1 - - - - - 2 - - - - - 3 - - - - - 4 - - - - - 5 - - - - - 6 - - - - - 7 - - - - - 8 - - - - - 9 - - - - - 10 - - - - - 11 + + + + + 12 + + + + + 13 + + + + + 14 + + + + + 15 + + + + + 16 + + + + + 17 + + + + + 18 + + + + + 19 + + + + + 20 + + + + +
9.0 Storage: 2 to 30oC
10.0 Expiration Dating: 24 months from date of manufacture
10.0 Method of Manufacture: 10.5 Nitrocellulose Membrane Manufacture: 10.5.1 The purified monoclonal antibody, diluted in phosphate buffer saline, is coated on the test region. Simultaneously, goat anti- mouse IgG polyclonal antibody, diluted in phosphate buffer saline, is coated on the control region. 10.5.2 The coated membrane is dried for a minimum of 24 hours then sealed in an aluminum bag which contains silica gel desiccant.
10.6 Anti-HCG colloidal gold conjugate pad manufacture: 10.6.1 A buffer solution containing mouse Anti-HCG monoclonal antibody/colloidal gold conjugate is coated onto non-woven cloth sheets. 10.6.2 The coated non-woven cloth is dried for minimum 24 hours then sealed in an aluminum bag which contains desiccant.
10.7 Test Device Assembly: 10.7.1 The coated membrane S1, the conjugate pad S4, and an absorbent pad is applied to an adhesive-coated backing. 10.7.2 A 2-part waterproof label is applied over the conjugate pad and the absorbent pad, the assembled sheet of material is cut into strips. The test strips are then vacuum-dried for a minimum of 4 hours. 10.7.3 The assembled test strip is sealed in a plastic cassette casing. The cassette device is sealed in an aluminum pouch along with a desiccant packet and disposable pipette. 10.8 Packaging available:
Product Code Test Medium Test Format Packing Store HCG 120 Urine Strip 1 x 100's HCG 121 Urine Cassette 1 x 40's bulk HCG 111 Urine Cassette 1 x 15's box HCG 110 Urine Strip 1 x 50's
11.0 Clinical Trials: To establish the sensitivity and specificity of Sensatest One-Step HCG test kit relative to other rates of qualitative urine HCG tests, 425 samples from women suspected of being pregnant were studied. The overall accuracy of Sensatest One-Step HCG test kit in this study was 100%, i.e., 425 samples got the expected results when confirmed with results from clinical follow ups. A commercially available qualitative test kit (ICON HCG) was used to compare with Sensatest One-Step HCG test kit for relative sensitivity and specificity in 803 urine samples. In total, 8 samples were discordant. In turn, the agreement is 99.0%. The results are shown in Table 3.
Table 3 - Comparison of One-step HCG with ICON product for 803 cases
Results of Sensatest kits Subtotal + - Results of ICON + 484 1 485 kits - 7 311 318 Subtotal 491 312 803
Based on quantitative test results on those discrepant samples (n = 8), Sensatest’s HCG test kits have better sensitivity and specificity. The results are shown in Table 4. All of these 8 discrepant samples were confirmed with clinical follow up 2 weeks later.
Table 4 - Quantitative results of discrepant samples Sample Quantitative results Sensatest ICON kits No. (mIU/ml) kits 1 25 + - 2 100 - + 3 28 - + 4 25 - + 5 40 - + 6 26 - + 7 50 - + 8 28 - +
12.0 Stability Studies: It is well known that immunochemicals are quite stable at room temperature if the materials are kept in dry condition. The chemicals used in the One-Step HCG device are dried on the membrane or chemical pad. The final test device is then sealed in an aluminum pouch with a desiccant. Accelerated stability testing has been carried out in this study to establish the shelf life and to predict the expiration date of the test.
Three lots of One-Step HCG test kit have been used for this study. The test devices in aluminum pouches were kept in the incubator at 42C or room temperature. The test kits were then removed from their storage and tested with positive and negative control panels. The results are shown in Table 5 and 6.
Table 5 – Results of Stability study on One-Step HCG test kits at 42C
HCG conc. 1 2 3 4 6 8 10 12 Lot # (mIU/ mo. mo. mo. mo. mo. mo. mo. mo. ml) 00501C ------0 00515C5 ------00515C4 ------00501C + + + + + + + + 20 00515C5 + + + + + + + + 00515C4 + + + + + + + + 00501C + + + + + + + + 100 00515C5 + + + + + + + + 00515C4 + + + + + + + +
Table 6 – Results of Stability study on One-Step HCG test kits at room temperature
HCG 1 2 4 6 8 10 12 14 16 18 24 conc. mo mo mo mo mo mo mo mo mo mo mo Lot # (mIU/ ml) 00501C ------0 00515C5 ------00515C4 ------00501C + + + + + + + + + + + 20 00515C5 + + + + + + + + + + + 00515C4 + + + + + + + + + + + 00501C + + + + + + + + + + + 100 00515C5 + + + + + + + + + + + 00515C4 + + + + + + + + + + + 1.0 Name of kit: SENSATEST One Step LH Test – Cassette
2.0 Indication: For In Vitro Detection of Ovulation - The one step LH Test is a qualitative test that can predict when there is a Luteinizing Hormone (LH) surge, and in turn, when one is likely to ovulate.
3.0 Package Size: One test per kit
4.0 Active Ingredients: 4.1 Coated Antibodies: 4.1.1 Control region: Goat anti-mouse(IgG) polyclonal antibody 4.1.2 Test region: Mouse monoclonal anti-LH antibody A 4.2 Labeled Antibodies: Colloidal gold conjugate of monoclonal anti-LH antibody B
11.0 Assay Protocol: 11.1 Assay Procedure: Read the package insert for Sensatest One-Step LH test carefully before starting the assay. Allow urine samples to warm up to room temperature before testing. 11.1.1 When you are ready to begin testing, open the sealed pouch by tearing along the notch. Remove the test device from its foil pouch. Place it on a flat surface and label it with patient ID. Use the device as soon as possible. 11.1.2 Using a pipette, dispense 3-4 drops (0.2 ml) of urine onto the sample well of the cassette. 11.1.3 Wait 10 minutes and read results. It is important that the background is clear before the result is read. Results obtained after 30 minutes should be considered invalid. 11.2 Interpretation of Results: 11.2.1 No LH Surge: Only one colour band appears on the control region or two bands appear and the test band is lighter than the control band. This indicates there is no LH surge. 11.2.2 LH Surge: Two colour bands are visible and the test band is equal to or darker than the control band, one will probably ovulate in the next 24-48 hours. If one wants to be pregnant, the best time to conceive is after 24 but before 48 hours. 11.2.3 Invalid: No visible band at all or no coloured line appears on the control region. Repeat test with a new test kit. 12.0 Specificity: 12.1 Cross Reactivity The cross reactivity of LH test kits was evaluated with LH homologous hormones. Homologous hormones FSH and TSH were added to urine samples containing LH at concentration of 0, 30 or 100 mIU/ml. No cross reactivity was observed in the study (shown as Table 1).
Table 1 - Cross-reactivity study of One-step LH test kit LH conc. in Un-spiked Urine samples spiked with homologous hormones urine sample urine FSH TSH (mIU/ml) samples 1000 mIU/ml 1000 mIU/ml - - - 0 ------+ + + 30 + + + + + + + + + 100 + + + + + +
12.2 Non-specific interference One-step LH test was checked for possible interference from visibly hemolyzed, lipemic and icteric samples. Human hemoglobin, bilirubin or albumin was spiked into samples with different concentration of LH and tested using un-spiked sample as controls. No significant interference was observed in 20 sample testing results that were either positive or negative for LH. The results are shown as Table 2.
Table 2 - Non-specificity study of One-step LH test kits Urine samples spiked with (mg/ml) Sample Un-spiked Hemoglobin Bilirubin Albumin No samples 10 1 0.06 100 1 - - - - - 2 - - - - - 3 - - - - - 4 - - - - - 5 - - - - - 6 - - - - - 7 - - - - - 8 - - - - - 9 - - - - - 10 - - - - - 11 + + + + + 12 + + + + + 13 + + + + + 14 + + + + + 15 + + + + + 16 + + + + + 17 + + + + + 18 + + + + + 19 + + + + + 20 + + + + + 13.0 Storage: 2 to 30C
14.0 Expiration Dating: 24 months from date of manufacture
10.0 Method of Manufacture: 10.9 Nitrocellulose Membrane Manufacture: 10.9.1 The purified monoclonal antibody, diluted in phosphate buffer saline, is coated on the test region. Simultaneously, goat anti- mouse IgG polyclonal antibody, diluted in phosphate buffer saline, is coated on the control region. 10.9.2 The coated membrane is dried for a minimum of 24 hours then sealed in an aluminum bag which contains silica gel desiccant. 10.10Anti-LH colloidal gold conjugate pad manufacture: 10.10.1 A buffer solution containing mouse Anti-LH monoclonal antibody/colloidal gold conjugate is coated onto non-woven cloth sheets. 10.10.2 The coated non-woven cloth is dried for minimum 24 hours then sealed in an aluminum bag which contains desiccant. 10.11Test Device Assembly: 10.11.1 The coated membrane S1, the conjugate pad S4, and an absorbent pad are applied to an adhesive-coated backing. 10.11.2 A 2-part waterproof label is applied over the conjugate pad and the absorbent pad, the assembled sheet of material is cut into strips. The test strips are then vacuum-dried for a minimum of 4 hours. 10.11.3 The assembled strip is placed in a plastic cassette casing. 10.11.4 The assembled cassette is sealed in an aluminum pouch along with a desiccant packet and disposable pipette. 10.12Packaging available :
Product Code Test Medium Test Format Packing Store
LH 131 Urine Cassette 1 x 50 11 Clinical Trials: To establish the sensitivity and specificity of Sensatest One-step LH test kit relative to other qualitative urine LH tests. 425 clinical samples were studied. The overall accuracy of Sensatest One-Step LH test kit in this study was 100%. Another commercially qualitative test kit (ABI LH) was used to compare with Sensatest One- Step LH test kit for relative sensitivity and specificity in 803 urine samples. In total, 8 samples were discordant, meaning the agreement rate is 99.0%. The detailed results are shown in Table 3.
Table 3 - Comparison of One-step LH with ABI product for 803 cases Subtotal Results of SENSATEST kits + - Results of ABI kits + 483 2 485 - 6 312 318 Subtotal 489 314 803
12 Stability Studies: It is well known that immunochemicals are quite stable at room temperature if the materials are kept in dry condition. The chemicals used in the One-Step LH 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 LH test kit have been used for this study. The test devices in aluminum pouches were kept in the incubator at 42C or room temperature. The test kits were removed from their storage and tested with positive and negative control panels. The results are shown in Table 4 and 5.
Table 4 – Results of Stability study on One-Step LH test kits at 42C LH conc. 1 2 3 4 6 8 10 12 Lot # (mIU/ml) mo. mo. mo. mo. mo. mo. mo. mo. 5-L00515 ------0 3-L00515 ------L00515NM ------5-L00515 + + + + + + + + 30 3-L00515 + + + + + + + + L00515NM + + + + + + + + 5-L00515 + + + + + + + + 100 3-L00515 + + + + + + + + L00515NM + + + + + + + + Table 5 – Results of Stability study on One-Step LH test kits at room temperature LH conc. 1 2 4 6 8 10 12 14 16 18 24 Lot # (mIU/ mo mo mo mo mo mo mo mo mo mo mo ml) 5-L00515 ------0 3-L00515 ------L00515NM ------5-L00515 + + + + + + + + + + + 30 3-L00515 + + + + + + + + + + + L00515NM + + + + + + + + + + + 5-L00515 + + + + + + + + + + + 100 3-L00515 + + + + + + + + + + + L00515NM + + + + + + + + + + + +: LH Surge -: No LH Surge
The data collected to date indicate that Sensatest Inc’s One-Step LH test kits is stable for at least 24 months 2- 30C.
13 Quality Control: 13.1 Appearance: All materials are visually inspected before their use in manufacturing. 13.2 Specificity: 10 test kits are randomly selected and tested with no LH Surge control panel. All test results must be negative. 13.3 Cut Off: 10 test kits are randomly selected and tested with a control panel that contains more than 30 mIU/ml LH. All test results must be positive. 13.4 Internal Control: A coloured band must appear in the control region of the membrane with each tested kit, which indicates proper performance and reagent reactivity. 13.5 Reproducibility: Randomly selected test kits from different lots must give the same results when assaying the same sample.
14 References: 14.1 Batzer FR, Fertility & Sterility, Vol 34, 1, 1980 14.2 Catt KI, Dufan ML & Vaitukaitis JL, Journal of Clinical Endocrinol. Metlab., Vol 40, 537, 1975 14.3 Braunstein GD, Rason J, Alder D, Danzer H, Wade ME, Am. J., Obstet. Gynecol., Vol 126, 677, 1976 14.4 Lenton E, Meal L, Sulaiman R, Fertility & Sterility, Vol 37, 773, 1982 14.5 Dawood M, Saeba B & Landesman R, Ob. Gyn., Vol 126, 678, 1976 14.6 Braunstein GD et al, Ann. Inter. Med., Vol 78, pp 39-45 14.7 Engvall E, Methods in Enzymology, Vol 70, pp 419-439, 1980 14.8 Uotila M, Ruoslahti E, Engvall E, J. Immun. Meth, Vol 42, 11, 1981