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72592.technicaldatasheet.pdf

Liofilchem® MYCOPLASMA SYSTEM Plus (Ref. 72592 - 79592) Rev.6 / 11.07.2012
MYCOPLASMA SYSTEM Plus
System for the detection, semi-quantitative determination and
susceptibility test of urogenital mycoplasmas
DESCRIPTION
MYCOPLASMA SYSTEM Plus
is a 24-well system containing desiccated biochemical and antibiotic substrates
for the detection, susceptibility test and semi-quantitative determination of urogenital mycoplasmas.
The system is inoculated with the suspension of the clinical sample and incubated at 36 ± 1 °C for 18-24 hours.
The tests used in the detection, count and susceptibility test are interpreted by evaluating the color of the various
wells and performing a microscope examination.
KIT CONTENTS
The kit contains:
20 systems MYCOPLASMA SYSTEM Plus
20 Vials di Physiological Solution (7 mL/vial) 4 systems MYCOPLASMA SYSTEM Plus
4 Vials di Physiological Solution (7 mL/vial) ITEMS NECESSARY BUT NOT INCLUDED IN THE KIT
Vaseline oil for microbiological use (VASELINE OIL 2 bottle of 50 mL) ref. 80278 CONFIGURATION
The configuration of the system is shown in Table 1.
Table 1
COUNT AND IDENTIFICATION OF MYCOPLASMAS / UREAPLASMAS
Growth of mycoplasmas (102 < titre < 104 CFU/mL) Growth of mycoplasmas (104 < titre < 105 CFU/mL) Growth of mycoplasmas (titre > 105 CFU/mL) DETECTION OF T.VAGINALIS AND CANDIDA spp.
6-TR/YE
Trichomonas vaginalis / Candida spp. SUSCEPTIBILITY TESTING OF THE MYCOPLASMAS / UREAPLASMAS
Liofilchem® MYCOPLASMA SYSTEM Plus (Ref. 72592 - 79592) Rev.6 / 11.07.2012
PRINCIPLE OF THE METHOD
MYCOPLASMA SYSTEM Plus
allows the detection, semi-quantitative count, presumptive identification and
susceptibility test of Mycoplasma hominis and Ureaplasma urealyticum isolated from clinical samples and the
detection and presumptive identification of the microorganisms most frequently isolated from vaginal and urethral
swabs and seminal fluid, such as Trichomonas vaginalis and Candida spp.
!
The semi-quantitative count of urogenital mycoplasmas (Mycoplasma hominis and Ureaplasma urealyticum) is
provided by the yellow to red color change of wells 1-GR+, 2-GR++, 3-GR+++.
The presence of Mycoplasma hominis is shown by the yellow to red color change of the Arginine Test well 4-ADC.
The presence of Ureaplasma urealyticum is shown by the yellow to red color change of the Urea Test well 5-UR.
The presence of Trichomonas vaginalis and Candida spp. is determined by microscope (40x) examination of a drop
of culture liquid taken from well 6-TR/YE evaluating the presence of mobile ciliated protozoites for identification of
Trichomonas vaginalis and/or the presence of chlamydospores and hyphae for identification of Candida spp.
The susceptibility testing of the urogenital mycoplasmas is evaluated by observing the color change in wells 7-TE
to 24-AZM as indicated in table 3.
COMPOSITION
Culture medium for growth of mycoplasmas Culture medium for growth of mycoplasmas Culture medium for growth of mycoplasmas Culture medium with substrate for decarboxylation of arginine Culture medium con with substrate for hydrolysis of urea 6-TR/YE
Culture medium for isolation of Trichomonas vaginalis and Candida spp. Culture medium containing Tetracycline - 4 μg/mL Culture medium containing Tetracycline - 8 μg/mL Culture medium containing Pefloxacin - 8 μg/mL Culture medium containing Pefloxacin - 16 μg/mL Culture medium containing Ofloxacin - 1 μg/mL Culture medium containing Ofloxacin - 4 μg/mL Culture medium containing Doxycycline - 4 μg/mL Culture medium containing Doxycycline - 8 μg/mL Culture medium containing Erythromicin - 8 μg/mL Culture medium containing Erythromicin - 16 μg/mL Culture medium containing Claritromycin - 8 μg/mL Culture medium containing Claritromycin - 16 μg/mL Culture medium containing Minociclyne - 4 μg/mL Culture medium containing Minociclyne - 8 μg/mL Culture medium containing Clindamycin - 4 μg/mL Culture medium containing Clindamycin - 8 μg/mL Culture medium containing Azithromycin - 4 μg/mL Culture medium containing Azithromicyn - 8 μg/mL Physiological Solution (g/L): Sodium chloride 9 g; Distilled water 1000 mL; pH 6.8 ± 0.2
Liofilchem® MYCOPLASMA SYSTEM Plus (Ref. 72592 - 79592) Rev.6 / 11.07.2012
COLLECTION AND CONSERVATION OF THE SAMPLES
Obtain the vaginal or urethral secretion with synthetic fibre swabs. Obtain the seminal fluid according to the
technique envisaged for the microbiological culture examination. The samples must be sent for inoculation in the
MYCOPLASMA SYSTEM Plus immediately after they are taken. Do not store in the refrigerator for even limited
periods of time, since low temperatures may damage the vitality of some particularly sensitive microorganisms
such as Trichomonas vaginalis prejudicing the final result.
TEST PROCEDURE
PREPARATION OF THE CLINICAL SAMPLE
Open one of the vials of Physiological Solution contained in the kit.
VAGINAL SWAB - URETHRAL SWAB
Immerse the swab (after obtaining the clinical material) in the vial of Physiological Solution and wait 5 minutes.
Carefully squeeze the swab against the vial wall so that the clinical material is dispersed homogeneously in thephysiological solution.
SEMINAL LIQUID
Add 0.2 mL of sample to the vial of Physiological Solution, shake and wait 5 minutes before inoculating thesystem.
Centrifuge 20 mL of urinary sample into two 10 mL tubes. Collect from each tube 0.1 mL of urinary sediment (0.2 mL)and add to the vial of Physiological Solution contained in the kit; shake and wait 5 minutes before inoculating thesystem.
CLINICAL MATERIAL FROM MYCOPLASMA TRANSPORT BROTH (ref. 20158)
Add 1 mL of Mycoplasma Transport Broth, containing the sample under examination, to the vial of PhysiologicalSolution, shake delicately and wait 5 minutes before inoculating.
INOCULATION OF THE SYSTEM
1. Take a system from its wrapper and allow to come to room temperature.
2. Write down the name of the patient, the date of the start of the examination, and the type of clinical material.
3. Transfer 0.2 mL (4 drops) of suspension of the clinical sample into each well of the system.
4. Cover all the wells except 6-TR/YE well with one drop of Vaseline oil for microbiological use.
5. Cover the system with the lid provided and incubate at 36 ± 1 °C for 18-24 hours. If the presence of
Mycoplasma hominis is suspected, incubate at 36 ± 1 °C for a further 24 hours.
INTERPRETATION OF THE RESULTS
At the end of the incubation:
!
Watch for the color change of the wells 1-GR+ to 5-UR and interpret the results using table 3 (COUNT AND
IDENTIFICATION OF THE MYCOPLASMAS / UREAPLASMAS).
Take a drop of liquid from well 6-TR/YE, deposit it on a glass slide, place a cover slip on top and watch for the
presence of Trichomonas vaginalis and Candida spp at the microscope (40x). Interpret as indicated in Table
No. 3 (SEARCH FOR T. VAGINALIS AND CANDIDA spp).
Watch for the color change of the wells from 6-TE to 24-AZM and interpret the results using Table No. 3
(SUSCEPTIBILITY TESTING OF THE MYCOPLASMAS / UREAPLASMAS). Refer to the well no. 1-GR+ as
growth control for the antibiogram of mycoplasma.
Note the results on the TEST RESULTS FORM (copy as many forms as necessary).
Liofilchem® MYCOPLASMA SYSTEM Plus (Ref. 72592 - 79592) Rev.6 / 11.07.2012
Well colour
COUNT AND IDENTIFICATION OF
MICOPLASMAS / UREAPLASMAS
Negative
Positive reaction
reaction
Growth of mycoplasmas (102 < titre < 104 CFU/mL) (1)
Growth of mycoplasmas (104 < titre < 105 CFU/mL) (2)
Growth of mycoplasmas (titre > 105 CFU/mL) (3)
Arginine Test: identifies Mycoplasma hominis Urea Test: identifies Ureaplasma urealyticum DETECTION OF T. VAGINALIS AND CANDIDA spp.
Microscopic observation (40x)
T. vaginalis: mobile ciliated trophozoite Trichomonas vaginalis / Candida spp. Candida spp: chlamydospores and hyphae SUSCEPTIBILITY TEST OF
Well colour
MYCOPLASMAS / UREAPLASMAS
S = Sensitive
I = Intermediate sensitivity
R = Resistant
LEGEND
(1) = equivalent to 5-20 colonies grown on Mycoplasma Agar A7
(2) = equivalent to 20-50 colonies grown on Mycoplasma Agar A7
(3) = over 50 colonies grown on Mycoplasma Agar A7
Liofilchem® MYCOPLASMA SYSTEM Plus (Ref. 72592 - 79592) Rev.6 / 11.07.2012
QUALITY CONTROL
Every batch of MYCOPLASMA SYSTEM Plus is subjected to quality control using the following reference
microorganisms:
FACTORS THAT MAY INVALIDATE THE RESULTS
Poor standardisation of the inoculum; application of the method to microorganisms not in the Mycoplasma group;
use of expired systems and/or reagents; incorrect application of the technique; non compliance with
temperatures and times of incubation.
LIMITS AND WARNINGS
The quality of the collecting influences the test result. A negative test, therefore, is not always index of absence
of infection. A positive test obtained with the system MYCOPLASMA SYSTEM Plus indicates an urogenital
mycoplasmas infection, but it cannot be used, alone, to make a clinical diagnosis.
PERFORMANCE
The results obtained with MYCOPLASMA SYSTEM Plus agree with those obtained using traditional culture
methods.
Application of the Wilcoxon non parametric statistical test shows that for each clinical material, the groups of
standard values and those obtained with MYCOPLASMA SYSTEM Plus do not differ significantly P ≤ 0.05 (1).
PRECAUTIONS
The product MYCOPLASMA SYSTEM Plus is not classified as hazardous under current legislation, however
see the safety data sheet for a correct use. MYCOPLASMA SYSTEM Plus is a disposable device to be used
only for diagnostic use in vitro. It must be used in the laboratory by properly trained personnel, using approved
aseptic and safety methods for handling pathogenic agents.
STORAGE
Store at 2-8 °C in the original packaging. Keep away from sources of heat and avoid excessive changes in
temperature. In such conditions, the product will remain valid until the expiry date indicated on the label.
Do not use beyond that date. Eliminate without using if there are signs of deterioration.
DISPOSAL OF USED MATERIAL
After use, MYCOPLASMA SYSTEM Plus and material that has come into contact with the sample must be
decontaminated and disposed of in accordance with the techniques used in the laboratory for decontamination
and disposal of potentially infected material.
PRESENTATION
Packaging
MYCOPLASMA SYSTEM Plus
MYCOPLASMA SYSTEM Plus
TABLE OF SYMBOLS

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