MATERIALS REQUIRED BUT NOT PROVIDED Timer For timing use.
Fetal Fibronectin Rapid Test PRECAUTIONS • For professional in vitro diagnostic use only. DEVICE RAPIDO Lateral Flow • Do not use after expiration date indicated on the package. Do not use the test if its foil pouch is damaged. Do not reuse tests. Tampone Cervico-Vaginale • This kit contains products of animal origin. Certified knowledge of the origin Product Code 500160 Packaging 20 test and/or sanitary state of the animals does not totally guarantee the absence of transmissible pathogenic agents. It is therefore, recommended that these For professional in vitro diagnostic use only. products be treated as potentially infectious, and handled observing the usual safety precautions (do not ingest or inhale). INTENDED USE • Avoid cross-contamination of specimens by using a new specimen collection container for each specimen obtained. The StrongStep® Fetal Fibronectin Rapid Test is a visually interpreted • Read the entire procedure carefully prior to performing any tests. immunochromatographic test intended to be used for the qualitative detection of • Do not eat, drink or smoke in the area where the specimens and kits are fetal fibronectin in cervicovaginal secretions. The presence of fetal fibronectin in handled. Handle all specimens as if they contain infectious agents. cervicovaginal secretions between 22 weeks, 0 days and 34 weeks, 6 days of Observe established precautions against microbiological hazards gestation is associated with elevated risk of preterm delivery. throughout the procedure and follow the standard procedures for proper INTRODUCTION disposal of specimens. Wear protective clothing such as laboratory coats, disposable gloves and eye protection when specimens are assayed. Preterm delivery, defined by the American College of Obstetricians and • Do not interchange or mix reagents from different lots. Do not mix solution Gynecologists as delivery prior to the 37th week of gestation, is responsible for bottle caps. the majority of non-chromosomal perinatal morbidity and mortality. Symptoms • Humidity and temperature can adversely affect results. of threatened preterm delivery include uterine contractions, change of vaginal • When the assay procedure is completed, dispose the swabs carefully after discharge,vaginal bleeding, backache, abdominal discomfort, pelvic pressure, autoclaving them at 121°C for at least 20 minutes. Alternatively, they can and cramping. Diagnostic modalities for identification of threatened preterm be treated with 0.5% sodium hypochloride (or house-hold bleach) for one delivery include uterine activity monitoring and performance of a digital cervical hour before disposal. The used testing materials should be discarded in examination, which allows estimation of cervical dimensions. These methods accordance with local, state and/or federal regulations. have been shown to be limited, as minimal cervical dilatation (< 3 centimeters) • Do not use cytology brushes with pregnant patients. and uterine activity occur normally and are not necessarily diagnostic of imminent preterm delivery. While several serum biochemical markers have STORAGE AND STABILITY been evaluated, none have been widely accepted for practical clinical use. Fetal fibronectin (fFN), an isoform of fibronectin, is a complex adhesive • The kit should be stored at 2-30°C until the expiry date printed on the glycoprotein with a molecular weight of approximately 500,000 daltons. sealed pouch. Matsuura and co-workers have described a monoclonal antibody called FDC-6, • The test must remain in the sealed pouch until use. which specifically recognizes III-CS, the region defining the fetal isoform of • Do not freeze. fibronectin. Immunohistochemical studies of placentae have shown that fFN is • Cares should be taken to protect components in this kit from contamination. confined to the extracellular matrix of the region defining the junction of the Do not use if there is evidence of microbial contamination or precipitation. maternal and fetal units within the uterus. Biological contamination of dispensing equipments, containers or reagents Fetal fibronectin can be detected in cervicovaginal secretions of women can lead to false results. throughout pregnancy by use of a monoclonal antibodybased immunoassay. Fetal fibronectin is elevated in cervicovaginal secretions during early pregnancy SPECIMEN COLLECTION AND STORAGE but is diminished from 22 to 35 weeks in normal pregnancies. The significance Use only Dacron or Rayon tipped sterile swabs with plastic shafts. It is of its presence in the vagina during the early weeks of pregnancy is not recommend to use the swab supplied by the kits manufacturer(The swabs understood. However, it may simply reflect the normal growth of the are not contained in this kit, for the ordering information, please contact the extravillous trophoblast population and the placenta. Detection of fFN in manufacturer or local distributor, the cataloge number is 207000). Swabs cervicovaginal secretions between 22 weeks, 0 days and 34 weeks, 6 days from other suppliers have not been validated. Swabs with cotton tips or gestation is reported to be associated with preterm delivery in symptomatic wooden shafts are not recommended. and between 22 weeks, 0 days and 30 weeks, 6 days in asymptomatic • Cervicovaginal secretions are obtained from the posterior fornix of the
pregnant women. vagina. The collection process is intended to be gentle. Vigorous or forceful
collection, common for microbiological cultures, is not required. During a PRINCIPLE speculum examination, prior to any examination or manipulation of the cervix or the vaginal tract, lightly rotate the applicator tip across the The StrongStep® fFN Test uses color immunochromatographic, capillary flow posterior fornix of the vagina for approximately 10 seconds to absorb technology. The test procedure requires the solubilization of fFN from a vaginal swab cervicovaginal secretions. Subsequent attempts to saturate the applicator by mixing the swab in Sample Buffer. Then the mixed sample buffer is added to the tip may invalidate the test. Remove the applicator and perform the test as test cassette sample well and the mixture migrates along the membrane surface. If directed below. fFN is present in the sample, it will form a complex with the primary anti- fFN • Put the swab to the extraction tube, if the test may be run immediately. If antibody conjugated to colored particles. The complex will then be bound by a immediate testing is not possible, the patient samples should be placed in a second anti- fFN antibody coated on the nitrocellulose membrane. The appearance dry transport tube for storage or transport. The swabs may be stored for 24 of a visible test line along with the control line will indicate a positive result. hours at room temperature (15-30°C) or 1 week at 4°C or no more than 6 month at -20°C. All specimens should be allowed to reach a room KIT COMPONENTS temperature of 15-30°C before testing. PROCEDURE 20 Individually packed Each device contains a strip with colored test devices conjugates and reactive reagents pre-coated Bring tests, specimens, buffer and/or controls to room temperature (15-30°C) at the corresponding regions. before use. 2 Extraction Buffer vial 0.1 M Phosphate buffered saline (PBS) and • Place a clean Extraction tube in the designated area of the workstation. 0.02% sodium azide. Add 1ml of Extraction Buffer to the extraction tube. 1 Positive control swab Contain fFN and sodium azide. For External • Put the specimen swab into the tube. Vigorously mix the solution by (on request only) control. rotating the swab forcefully against the side of the tube for least ten 1 Negative control swab Not contain fFN. For external control. times (while submerged). Best results are obtained when the specimen is (on request only) vigorously mixed in the solution. 20 Extraction tubes For specimens preparation use. • Squeeze out as much liquid as possible from the swab by pinching the 1 Workstation Place for holding buffer vials and tubes. side of the flexible extraction tube as the swab is removed. At least 1/2 of 1 Package insert For operation instruction. the sample buffer solution must remain in the tube for adequate capillary migration to occur. Put the cap onto the extracted tube. Discard the swab in a suitable biohazardous waste container. • The specimens extracted can retain at room temperature for 60 minutes Analytic sensitivity without affecting the result of the test. • Remove the test from its sealed pouch, and place it on a clean, level The lowest detectable amount of fFN in the extracted sample is 50μg/L. surface. Label the device with patient or control identification. To obtain ≥ μ -7 a best result, the assay should be performed within one hour. Among symptomatic women, elevated levels ( 0.050 g/mL) (1 x 10 • Add 3 drops (approximately 100 µl) of extracted sample from the mmol/L) of fFN between 24 weeks, 0 days and 34 weeks, 6 days indicate Extraction Tube to the sample well on the test cassette. increased risk of delivery in ≤ 7 or ≤ 14 days from sample collection. Among Avoid trapping air bubbles in the specimen well (S), and do not drop any asymptomatic women, elevated levels of fFN between 22 weeks, 0 days and solution in observation window. 30 weeks, 6 days indicate increased risk of delivery in ≤ 34 weeks, 6 days of As the test begins to work, you will see color move across the membrane. • Wait for the colored band(s) to appear. The result should be read at 5 gestation. The cutoff of 50 μg/L fFN was established in a multicenter study minutes. Do not interpret the result after 5 minutes. conducted to evaluate the association between fetal fibronectin expression Discard used test tubes and Test Cassettes in suitable biohazardous waste during pregnancy and preterm delivery. container. Interfering Substances INTERPRETATION OF RESULTS Care must be taken not to contaminate the applicator or cervicovaginal secretions with lubricants, soaps, disinfectants, or creams. Lubricants or POSITIVE RESULT: Two colored bands appear on the membrane. One creams may physically interfere with absorption of the specimen onto the C band appears in the control region (C) and another applicator. Soaps or disinfectants may interfere with the antibody-antigen T band appears in the test region (T). reaction. Potential interfering substances were tested at concentrations that might be NEGATIVE RESULT: reasonably found in cervicovaginal secretions. The following substances did not Only one colored band appears in the control region interfere in the assay when tested at the levels indicated. C (C). No apparent colored band appears in the test Substance Concentration Substance Concentration T region (T). Ampicillin 1.47 mg/mL Prostaglandin F2 0.033 mg/mL Erythromycin 0.272 mg/mL Prostaglandin E2 0.033 mg/mL Control band fails to appear. Results from any test Maternal Urine 5% (vol) MonistatR 0.5 mg/mL INVALID RESULT: which has not produced a control band at the 3rd Trimester (miconazole) specified reading time must be discarded. Please C C Oxytocin 10 IU/mL Indigo Carmine 0.232 mg/mL review the procedure and repeat with a new test. If Terbutaline 3.59 mg/mL Gentamicin 0.849 mg/mL T T the problem persists, discontinue using the kit Dexamethasone 2.50 mg/mL BetadineR Gel 10 mg/mL immediately and contact your local distributor. MgSO4•7H2O 1.49 mg/mL BetadineR 10 mg/mL Cleanser NOTE: Ritodrine 0.33 mg/mL K-YR Jelly 62.5 mg/mL 1. The intensity of the color in test region (T) may vary depending on the DermicidolR 25.73 mg/mL concentration of aimed substances present in the specimen. But the 2000 substances level can not be determined by this qualitative test. 2. Insufficient specimen volume, incorrect operation procedure, or performing expired tests are the most likely reasons for control band failure. LITERATURE REFERENCES 1. American College of Obstetricians and Gynecologists. Preterm Labor. Technical Bulletin, Number 133, October, 1989. QUALITY CONTROL 2. Creasy RK, Resnick R. Maternal and Fetal Medicine: Principles and Practice. • Internal procedural controls are included in the test. A colored band Philadelphia: W.B. Saunders; 1989. appearing in the control region (C) is considered as an internal positive 3. Creasy RK, Merkatz IR. Prevention of preterm birth: clinical opinion. Obstet procedural control. It confirms sufficient specimen volume and correct Gynecol 1990;76(Suppl 1):2S–4S. procedural technique. 4. Morrison JC. Preterm birth: a puzzle worth solving. Obstet Gynecol • External procedural controls may provided(on request only) in the kits to 1990;76(Suppl 1):5S-12S. ensure that the tests are functioning properly. Also, the Controls may be 5. Lockwood CJ, Senyei AE, Dische MR, Casal DC, et al. Fetal fibronectin in used to demonstrate proper performance by the test operator. To perform a cervical and vaginal secretions as a predictor of preterm delivery. New Engl J positive or negative control test, complete the steps in the Test Procedure Med 1991;325:669–74. section treating the control swab in the same manner as a specimen swab. GLOSSARY OF SYMBOLS LIMITATIONS OF THE TEST Catalog number Temperature limitation 1. This assay can only be used for the qualitative detection of fetal fibronectin in cervicovaginal secretions. Consult instructions for use Batch code 2. Test results should always be used in conjunction with other clinical and In vitro diagnostic medical Use by laboratory data for patient management. device 3. Specimens should be obtained prior to digital examination or manipulation Contains sufficient for
7. The performance characteristics of the StrongStep® fFN test are based on studies in women with singleton gestations. Performance has not been verified on patients with multiple gestations, e.g., twins. 8. The StrongStep® fFN test is not intended to be performed in the presence of rupture of amniotic membranes and rupture of amniotic membranes should be ruled out prior to conducting the test. PERFORMANCE CHARACTERISTICS ) Table: StrongStep® fFN Test vs. Another brand fFN Test
Relative Sensitivity: Another brand 97.96% (89.13%-99.95%)* + - Total Relative Specificity: 98.73% (95.50%-99.85%)* StrongStep® + 48 2 50 Overall Agreement: fFN Test 98.55% (95.82%-99.70%)* - 1 156 157 *95% Confidence Interval 49 158 207