1. Purpose

The kit is for rapid diagnosis and treatment evaluation of Bacterial vaginosis on the basis of 4 tests (multi-test), which includes H₂O₂ determination, sialidase activity test, leukocyte esterase activity test and pH value measurement.

2. Outline

Bacterialvaginosis is actually a symptom-comlex caused by the decrease of common bacterial population and the increase of anaerobic bacteria, e.g., Bacteroides, Mobiluncus spp, Gardnerella vaginalis, Prevotella spp, Peptococeus, and Mycoplasma hominis, etc. With a high morbidity (10-15%), Bacterial vaginosis is often attributed to habitual abortion, premature labor, premature breaking of fetal membrane, intrauteral infection, pelvic inflammation, cervical ulcer, andsome other complicateddiseases. The polybacterial infection yields great difficulty to bacteriological diagnosis. The standard BV laboratory diagnostic method "Amsel" , so called gold standard, which is well recognized, covers the following items : (1). Vagnial secretion pH>4.5 (2).Increased thin vaginal secretion with peculiar smell (3). Positive amine test (4). The clue cell can be found in vaginal secretion smear. The card is characterized by accuracy, simplicity, rapidity and sensitivity. The correlation rate with Amsel reaches 92%, providing a satisfactory diagnostic result.

3. Working Principle

The kit is scientifically prepared on the basis of multi-test of pH, H₂O₂,sialidase and leuk-esterase. The operation of the test kit is very simple. The whole test can be done within 15 to 20 minutes and needs no special apparatus. What the kit assays is the related enzymes leading to Bacterial vaginosis or vaginitis.

(1).H₂O₂ determination

H₂O₂ reacts with peroxidase to produce nascent oxygen. With existence of 4-Aminoantipyrine,

The nascent oxygen oxidizes 2,4,6-Trihydroxybenzoic acid to show red. The red color grade is proportionable to H₂O₂ concentration.

(2). Sialidase activity test

The substrate BCIN (5-Bromo-4-chloro-3-indolyl -α-D-N-acetylneuraminic acid sodium), while reacting with neuramidinase (sialidase), will be decomposed into indoles, which will be oxidized to show blue. The blue color grade is proportionable to sialidase activity.

(3). Leukocyte esterase test

The substrate BCIA (5-Bromo-4-chloro-3-indolyl -α-D-N-acetyl acetate), while reacting with leukocyte esterase, will be decomposed into indoles, which will be oxidized to show blue. The blue color grade will be proportion able to leukocyte esterase activity.

(4). pH value measurement

The test is based on pH indicator and pH paper. Yellow, yellowish green or green indicate pH ≤4.5, whereas bluish green or blue indicate pH>4.5

4. Contents of the kit

(1) BV test card 20/40pcs

(2) BV extracting Liquid 12ml×1 btl

(3) Chromogenic reagent 3ml×1 btl

(4) Dropper 20/40pcs

(5) Lysis tube 20/40pcs

(6) Instruction 1pcs

5. Attentions

(1) Read the instruction carefully before operation.

(2) Deal with the waste according to regulations of hospitals or the state environmental protection departments because of some potential risk.

(3) Don’t damp the swab with saline before sampling, so as not to affect the pH test.

(4) For in vitro use only.

6. Sample collection

Insert Collect secretion with a swab and vaginal speculum from the vaginal posterior fornix. Sample should be sufficient and sent to test immediately.

7. Operation procedures

(1) Open a test card, first scrub the sample 2-3 times in the pH test slot, observe the color change and record pH value;

(2) Put sample swab into lysis tube and add 8-10 drops (appx. 350~400μl) of extracting liquid, squeeze the swab repeatedly in order to get a thorough extraction;

(3) Add the extracted fluid with a transfer pipet to other three test slots (for H₂O₂, SA and LE respectively), 2 drops each;

(4) Add a drop of chromogenic reagent to the H₂O₂ test slot;

(5) Place the test card in Encode made incubator and keep at 42~45°C for 15min;

Attention: The result to read after 30min will be invalid.

8. Observation of results

(1) pH: Yellow, yellowish green or green is normal, bluish-green or blue is abnormal.

(2) H₂O₂: Red is normal, no color change is abnormal.

(3) SA: Light blue, blue is positive, no color change is negative.

(4) LE: Bluish green, blue light blue is positive, no color change is negative.

9.Reference for the Multi-Test results

10. Detection limit

The detection limit of H₂O₂ is 2μmol/L, SA 7 units per ml, LE 9 units per ml.

11. Storage and validity duration

12 months at 2~8°C.