Keul-o-test

Vaginitis Multitest

KGST0106

Intended Use
Keul-o-test vaginitis multitest is intended for the detection of the 5 indicators of vaginitis which are Lactobacillus, leukocyte, bacterial vaginosis(BV), Trichomoniasis vaginitis(TV), Candida vaginitis (VVC).

Introduction
The product can diagnose female vaginal diseases, including Lactobacillus, leukocyte, bacterial vaginosis(BV), trichomoniasis vaginitis(TV), candida vaginitis(VVC) by detecting pH, hydrogen peroxide, leukocyte esterase, neuraminidase, proline aminopeptidase and N-acetyl-p-D-glucosaminidase. Because sampling the vagina biochemical markers is simple, repeatable, objective, fast, cheap and does not need special equipment, it's so convenient for clinical application. It can diagnose suspected vaginal infections, find out the infected asymp­tomatic individual, investigate the conventional treatment-resistant cases, help select clinical treatment and judge the cure as well as the vaginal infection prevalence survey. It is a gyne­cologic diagnostic technology with a bright future, and is highly valuable in the gynecologic diagnostic field.

Biological Principle of the Procedure
This kit is based on the chromogenic phenomenon when certain specific substrates are de­composed by an enzyme, or, in the case of non-chromogenic reactions, with the assistance of a chromogenic substrate. Under the catalysis of peroxidase, hydrogen peroxide reacts with its corresponding substrate and the substrate turns red. The color intensity is directly proportional to the concentration of H₂O₂. When the vaginal inflammation occurred, leukocyte esterase ac­tivity in vaginal secretions would significantly increase. Because leukocyte esterase hydrolyses a specific substrate, when the sample contains the enzyme, a biochemical reaction would occur leading to the display of a blue color and the color intensity is directly proportional to the es­terase activity. When neuraminidase is present in the sample, it reacts with the specific sub­strate, upon the addition of a chromogenic reagent, a red or purple color will display and the color intensity is directly proportional to the neuraminidase activity. Proline aminopeptidase specificly hydrolyses its corresponding substrate and lead to the display a yellow color and the color intensity is inversely proportional to the proline aminopeptidase activity; N-acetyl-p-D-glucosaminidase hydrolyses a corresponding substrate to display a yellow color under alkaline conditions and the color intensity is directly proportional to the N-acetyl-p-D-glucosaminidase activity.

Components

1. Vaginitis Cassette
   20 cassettes each containing 6 wells
2. Sample Diluent
   1 bottle containing 13.0 ml of sample diluent
3. Chromogenic Reagent
   1 bottle containing 1.5 ml of chro-mogenic reagent
4. Stop Solution
   1 bottle containing 1.5 ml of stop solution
5. Color Reference
   1 sheet of color reference card
6. Sample Tube
   20 tubes
7. Dropper
   20 droppers

Materials Required but not Provided

1. Incubator

Warnings and Precautions

1. This product is for in vitro diagnostic use only and has been granted a state utility model patent. Patent No.: ZL 200720089307.9
2. Do not use reagents after expiry date.
3. Do not mix or use components from kits with different lot numbers.
4. This assay should be operated by a professional under the room temperature of 1 5-30°C and a relative humidity of 45-85%. After operation, the rest of the reagents should be stored at 2-8C .
5. Only test fresh samples. Avoid sexual intercourse periods and the menstruation period.
6. When removing the lid on the Vaginitis Cassette please pay attention not to get the lid and the reagent paper sticked together since this would drag the reagent paper out.
7. Samples with blood affect test results; it should be used with caution.
8. Chromogenic Reagent should only be applied to the well marked with NA, the Stop Solution should only be applied to the well marked with NAG.
9. Avoid any skin contact with all reagents. Stop solution contains NaOH. In case of contact, wash thoroughly with water.
10. Please immediately note down the name or number of the subject at the matte surface behind the "No." mark.
11. This product is for single use only. Samples and used items should be disposed of in ac­cordance with the infectious diseases laboratory practices.
12. Do not smoke, drink, eat or apply cosmetics in the working area. Use protective clothing and disposable gloves.
13. Time limits for reading each well when result is read with naked eyes. Read the first well (H2O2) within 5 minutes, the second well (LE) within 2 minutes, the third well (NA) within 5 minutes, the fourth well (PIP) within 5 minutes, the fifth well (NAG) between 30 seconds and 5 minutes of adding Stop Solution and sixth well (pH) within 1 minute.
14. When the accompanying instrument is used to read the result, add samples to the pH well after incubation rather than before incubation.

Storage Instructions
Unopened kits should be stored at 2-8°C and Vaginitis Cassettes should be kept sealed before use, and be used within 24 hours once unpacked. This cassette may be used throughout the expiry date of the kit (12 months from the date of manufacture). Refer to the package label for the expiry date.

Specimen Collection and Preparation

1. The sample is the secretion from vaginal vault or posterior viginal which can be taken with a sterile swab. The sample should be tested immediately.
2. When collecting the samples, keep turning around the swab at the vaginal vault or the posterior viginal for 10 - 20 seconds, until it is clearly seen that the secretion has been attached onto the swab.

Reagent Preparation

1. All reagents should be brought to room temperature (18-25°C) and leave for 30 minutes prior to use.
2. Adjust the incubator at 37°C.

Assay Procedure

• When result is read with naked eyes
1. Add 6-8 drops of Sample Diluent into Sample Tube. Place the sample swab in Sample Tube and wash thoroughly. After washing, gently press the wall of the Sample Tube to allow the solution adsorbed in the swab reflux into the tube. Discard the swab and retain the sample solution.
2. Unwrap a piece of Vaginitis Cassette and gently remove the lid. Add 1 drop of the sample solution into each well with a dropper. Then add 1 drop of Chromogenic Reagent into the well marked with NA. Gently shake Vaginitis Cassette horizontally. Within 1 minute, read the pH value according to the Color Reference.
3. Incubate Vaginitis Cassette at 37±1 °C for 1 5 minutes. Then add a drop of Stop Solution to the well marked with NAG. Read the results for the remaining 5 wells between 30 seconds and 2 minutes.
   
• When result is read with the accompanying instrument
1. Add 6-8 drops of Sample Diluent into Sample Tube. Place the sample swab in Sample Tube and wash thoroughly. After washing, gently press the wall of the Sample Tube to allow the solution adsorbed in the swab reflux into the tube. Discard the swab and retain the sample solution.
2. Unwrap a piece of Vaginitis Cassette and gently remove the lid. Add 1 drop of the sample solution into each of the firsts 5 wells with a dropper. Then add 1 drop of Chromogenic Reagent into the well marked with NA. Gently shake Vaginitis Cassette horizontally.
3. Incubate Vaginitis Cassette at 37±1 C for 1 5 minutes. Then add a drop of Stop Solution to the well marked with NAG. Afterwards, add a drop of sample solution to the well marked with pH.
4. Place Vaginitis Cassette in the accompanying instrument.
5. Read the result with the software provided and print out the test report from within the software.

Results (for result read with naked eyes)

1. The first well on the Vaginitis Cassette (Hydrogen Peroxide, H₂O₂)
   Colorless or light yellow is positive (indicated "+"), meaning the existance of disorder in vaginal flora. This reprensents a sick vaginal environment or a sub-health state; Light red (indicated "±") means the presence of a medium amount of Lactobacillus. In this case, the patient should be re-diagnosed clinically but this is usually regarded as negative and the vaginal flora display a trend of abnormaty or is in the recovery period; red or purple-red (indicated "-") is negative, meaning the possible existence of a large number of Lactobacillus. The vaginal flora is healthy in this case.
   
2. The second well on the Vaginitis Cassette (Leukocyte esterase, LE)
   colorless or a light color is negative (indicated "-"), meaning the concentration of leukocyte is less than 5 leukocytes/Hp; light blue is negative (indicate "±"), meaning the concentration of leukocyte is 5~1 5 leukocytes/Hp; blue is positive (indicated "+" "++","+ ++"), meaning the concentration of leukocyte is more than 1 5 leukocytes/Hp.
   
3. The third well on the Vaginitis Cassette (Neuraminidase, NA)
   Colorless or orange is negative (indicated "-"); light red is weak positive (indicated "±"); red, purple-red, blue, brown or black is positive (indicated "+"). A positive reading in this well means the infection of bacterial vaginosis (BV).
   
4. The fourth well on the Vaginitis Cassette (Proline Aminopeptidase, PIP)
   Colorless or a light color is negative (indicated "-"); light yellow is weak positive (indicated "±"); yellow is positive (indicated "+"). The concurrence of positive reading in this well and negative reading in the 5th well means the patient is probably infected with bacterial vaginosis (BV). If both readings in the 4th and 5th are positive. The judgement should be based on the result of the 5th well.
   
5. The fifth well on the Vaginitis Cassette (Acetyl Glucosaminidase, NAG)
   Colorless or a light color is negative (indicate "-"); light yellow is weak positive (indicate "±"); yellow is positive (indicated "+"). If this well is positive, and the pH>4.8 at the same time, the subject is infected with trichomoniasis vaginitis (TV). Otherwise, if the pH<4.6, the subject is infected with Candida vaginitis (VVC).
   
6. The sixth well on the Vaginitis Cassette (pH)
   The transition of color from yellow, cyan through to green, blue represents the transition of pH from 3.8 to 5.4. according to the color reference, yellow and green represent the pH range is <4.6 while blue represents the pH range is >4.8.

Limitations of the Procedure
The detection principle is based on the principle of biochemical reaction. The improper sam­pling could lead to false result. In addition, female vaginal flora is complex and many interfere factors exist. For diagnostic purposes, the results obtained from this assay should always be used in combination with the clinical examination, patients' medical history, as well as other findings.

References

1. Smitka CM, and Jackson SG. 1989. Rapid fluorogenic assay for differentiation of the Can­dida parapsilosis group from other Candida spp.. J Clin Microbiol, 27(1):203-206.
2. James JN,Thomason JL,Gelbart SM,Osypowski P, and Hanson L.1992. Is trichomoniasis ofen associated with bacterial vaginosis in pregnant adolescents?. Am J Obstet Gynecol, 166(3):859-863.
3. Schoonmaker JN, Lunt BD, Lawellin dw, French JI, Hillier SL, and McGreqor JA. 1991. A new proline aminopeptidase assay for diagnosis of bacterial vaginosis. Am J Obstet Gynecol, 165(3):737-742.
4. Briselden AM, Moncla BJ, Stevens CE, and Hillier SL. 1992. Sialidases (neuraminidases) in bacterial vaginosis and bacterial vaginosis-associated microflora. J Clin Microbiol, 30(3):663-666.

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