BIOMÉRIEUX

422083 050923-02 - IVD - en - 2020-03

MYCOPLASMA IST 3
Diagnosis of urogenital mycoplasma (culture, identification, indicative enumeration, susceptibility testing)

INTENDED USE
MYCOPLASMA IST 3 is a manual qualitative and semi-quantitative in vitro diagnostic test for the culture, identification,
indicative enumeration and antibiotic susceptibility testing of Mycoplasma hominis and Ureaplasma spp. including
Ureaplasma parvum and Ureaplasma urealyticum.
It is intended as an aid to diagnosis and prediction of treatment response in adults suspected of having urogenital
mycoplasma infection, using urethral swabs, vaginal and cervical swabs, semen and male urine samples.
This assay is intended for use in clinical laboratories by laboratory health professionals.

PRINCIPLE
MYCOPLASMA IST 3 combines a selective culture broth with a strip containing 25 test cupules.
The broth provides optimum growth conditions for Mycoplasma (pH, substrates, association of several growth factors).
If a culture broth is positive, then specific substrates and phenol red indicator present in the broth (urea for
Ureaplasma spp. and arginine for M. hominis) will change color, due to an increase in pH.
The combination of three antibiotics and one antifungal agent provides selectivity, ensuring that any contaminating flora
present in the specimen does not affect the test.
After inoculation, the broth is dispensed into the strip.
This strip provides simultaneous results for:
- detection/identification,
- indicative enumeration,
- susceptibility testing with 4 antibiotics for M. hominis and 5 antibiotics for Ureaplasma spp.
MYCOPLASMA IST 3 can be used with mixed-culture specimens.

CONTENT OF THE KIT (25 TESTS)

- 25 MYCOPLASMA R1 vials containing 3.1 mL of broth (R1)
- 25 MYCOPLASMA R2 vials containing 1 mL of lyophilized broth (R2)
- 25 MYCOPLASMA IST 3 strips (STR)
- 25 incubation lids (INCUB)
- 1 package insert provided in the kit or downloadable from www.biomerieux.com/techlib

COMPOSITION AND RECONSTITUTION OF THE KIT REAGENTS

1. MYCOPLASMA R1 vial
Each vial contains 3.1 mL of broth, incorporating the stable nutrients required for specimen preparation. It inhibits most
Gram-positive and Gram-negative bacteria and is used to reconstitute the MYCOPLASMA R2 reagent.
2. MYCOPLASMA R2 vial
Each vial contains 1 mL of lyophilized Urea-Arginine broth. After reconstitution of MYCOPLASMA R2 with 3 mL of
MYCOPLASMA R1, the composition is the following:

Composition of the Urea-Arginine LYO 2 broth (MYCOPLASMA R1 + MYCOPLASMA R2)

Theoretical formula
This medium has been adjusted and/or supplemented according to the performance criteria required:
Meat peptone (porcine or bovine) 8g
Casein peptone (bovine) 8g
Yeast extract 4g
Sodium chloride 3.5 g
Arginine hydrochloride 5g
Cysteine hydrochloride 0.1 g
Urea 1g
Phenol red 0.05 g
PolyViteXTM mixture 10 mL
Horse serum 100 mL
Antibiotic mixture 10 mL
Purified water 1L
pH 6.3*
* The pH of the Urea-Arginine LYO 2 broth may decrease to as low as 6.0 over time. This decrease does not affect product performance in any way.

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3. MYCOPLASMA IST 3 strip

The strip contains 25 test cupules.
3.1. Detection/Identification (3 cupules)
- 0: growth control
- Uspp: identification of Ureaplasma spp.
- Mh: identification of M. hominis
Cupules Tests Principal substrates
0 0 Phenol red (0.05 g/L)
(growth control)
Uspp Uspp Phenol red (0.05 g/L)
Lincomycin
Mh Mh Erythromycin

3.2. Indicative enumeration (5 cupules)

These tests determine whether the mycoplasma count in the specimen is greater than or equal to different thresholds:
103,104, 106 CFU/mL (Colony Forming Unit):
- Uspp ≥ 103: Ureaplasma spp. titer ≥ 103 CFU/mL in the specimen.
- Uspp ≥ 104: Ureaplasma spp. titer ≥ 104 CFU/mL in the specimen.
- Uspp ≥ 106: Ureaplasma spp. titer ≥ 106 CFU/mL in the specimen.
- Mh ≥ 104: M. hominis titer ≥ 104 CFU/mL in the specimen.
- Mh ≥ 106: M. hominis titer ≥ 106 CFU/mL in the specimen.
Organism Cupules Tests Principal substrates
≥ 103 Uspp ≥ 103 Phenol red (0.05 g/L)
Ureaplasma spp. ≥ 104 Uspp ≥ 104 Lincomycin
Inhibition agent
≥ 106 Uspp ≥ 106
≥ 104 Mh ≥ 104 Erythromycin
M. hominis
≥ 106 Mh ≥ 106 Inhibition agent

3.3. Susceptibility tests (17 cupules)

Concentrations
Organism Cupules Antibiotics and
Abbreviations mg/L

LVX Levofloxacin LVX 2 4

MXF Moxifloxacin MXF 2 4
a
Ureaplasma spp. TET Tetracycline TET 1 2
ERY Erythromycin ERY 8 16
TEL Telithromycin TEL 4
LVX Levofloxacin LVX 1 2

b MXF Moxifloxacin MXF 0.25 0.5

M. hominis
TET Tetracycline TET 4 8
CLI Clindamycin CLI 0.25 0.5
a
Each susceptibility test cupule contains Lincomycin.
b
Each susceptibility test cupule contains Erythromycin.

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REAGENTS AND MATERIALS REQUIRED BUT NOT PROVIDED

Reagents
 Mineral oil (Ref. 70100)
 Urea Arginine LYO 2 (Ref. 42508)
 NaCl 0.85%
Materials
 ATBTM Electronic Pipette, ATB™ VIAFLO Pipette (or equivalent): consult bioMérieux
 Specimen collection swabs or ESwab®
 Pipettes and micropipettes
 Bacteriology incubator

WARNINGS AND PRECAUTIONS

 For in vitro diagnostic use and microbiological control.
 For professional use only. This test is intended for use by trained laboratory professionals.
 This kit contains products of animal origin. Certified knowledge of the origin and/or sanitary state of the animals does
not totally guarantee the absence of transmissible pathogenic agents. It is therefore recommended that these
products be treated as potentially infectious, and handled observing the usual safety precautions (do not ingest; do
not inhale).
 All specimens, microbial cultures and inoculated products should be considered infectious and handled appropriately.
Aseptic technique and usual precautions for handling the bacterial group studied should be observed throughout this
procedure. Refer to "CLSI® M29-A Protection of Laboratory Workers From Occupationally Acquired Infections;
Approved Guideline - Current revision". For additional handling precautions, refer to "Biosafety in Microbiological and
Biomedical Laboratories - CDC/NIH – Latest edition", or to the regulations currently in use in each country.
 The media should not be used as manufacturing material or components.
 Do not use reagents after the expiry date.
 Allow reagents to come to room temperature before use.
 Before use, check that the vial cap is intact.
 Do not use reagents if the packaging is damaged.
 Do not use vials with a turbid appearance.
 Do not use strips which have been damaged: for example, cupules deformed, desiccant sachet open.
 Interpretation of the test results should be made taking into consideration the patient’s history, the source of the
specimen, the colonial and microscopic morphology of the strain and, if necessary, the results of any other tests
performed, particularly the antimicrobial susceptibility patterns.
 Use of the test may be difficult for people who have problems recognizing colors.
 Only use the Urea-Arginine LYO 2 (MYCOPLASMA R1 + MYCOPLASMA R2) broth provided in the MYCOPLASMA
IST 3 kit to inoculate strips from the same MYCOPLASMA IST 3 kit.
 The antibiotics are tested on the specimen without taking into account the mycoplasma titer of the specimen. An
inoculum effect may be observed. In the case of low titers, the real susceptibility of the strain may be different from
the result obtained with the strip.

STORAGE CONDITIONS
The strips and vials should be stored in their box at +2°C/+8°C until the expiry date.

SPECIMEN COLLECTION AND PREPARATION

This kit can be used with the following specimens:
- Urethral swab
- Cervicovaginal swab
Since Mycoplasma have a high affinity for mucus cell membranes, it is important to thoroughly scrape the mucosa so
as to collect as many cells as possible.
Calcium alginate, Dacron or polyester swabs with aluminium or plastic shafts are preferred. Wooden shaft cotton
swabs should be avoided because of potential inhibitory effects (12).
- Semen
- Male urine (first-void)
Collect the specimen before administering any antibiotic treatment.
Note that a treatment containing beef bile or sertaconazole nitrate could act as an inhibitor for Mycoplasma.
A standardized technique must be used to prevent contamination by other microorganisms.
Good laboratory practices for specimen collection should be respected and adapted to the detection of Mycoplasma.

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INSTRUCTIONS FOR USE

Specimen Processing and Inoculum Preparation
CAUTION! Only use the Urea-Arginine LYO 2 (MYCOPLASMA R1 + MYCOPLASMA R2) broth provided in the
MYCOPLASMA IST 3 kit to inoculate strips from the same MYCOPLASMA IST 3 kit.
1. Allow the vial of MYCOPLASMA R1 to come to room temperature.
2. After collection,
- If using a dry swab, rapidly release the sample in the MYCOPLASMA R1 solution.
- If using an ESwab®, transfer 200 µL of the liquid transport medium into the MYCOPLASMA R1 solution.
- If using liquid samples, transfer 200 µL into the MYCOPLASMA R1 solution.
- If the volume of the liquid sample is < 200 µL but ≥ 100 µL, reconstitute MYCOPLASMA R2 with the
MYCOPLASMA R1 solution. Then adjust the Urea-Arginine LYO 2 broth (MYCOPLASMA R1 solution +
MYCOPLASMA R2 solution) proportionally to the sample to obtain the same proportion between the liquid sample
and the Urea-Arginine LYO 2 broth. Go directly to step 5 of this procedure.
3. Ensure that the maximum storage time given below, depending on the temperature, is respected:

Temperature +18°C/+25°C +2°C/+8°C

Maximum storage time

for inoculated
20 hours 72 hours
MYCOPLASMA R1
solution

False results may be obtained if these conditions are not respected.

4. After mixing, transfer 3 mL of the inoculated MYCOPLASMA R1 solution into the vial of MYCOPLASMA R2.
5. Mix using a vortex-type mixer to ensure that the lyophilization pellet is completely dissolved.
Strip Preparation and Incubation
1. Allow the strip to come to room temperature.
2. Remove the strip from its packaging.
3. Discard the desiccant.
4. Record the specimen collection reference on the elongated flap of the strip. (Do not record the reference on the lid as
it may be misplaced during the procedure).
5. Immediately dispense 55 µL of the Urea-Arginine LYO 2 (MYCOPLASMA R1 + MYCOPLASMA R2) broth into each
of the 25 test cupules on the MYCOPLASMA IST 3 strip using the ATBTM Electronic Pipette or the ATB™ VIAFLO
Pipette (or equivalent).
6. Add 2 drops of mineral oil to each cupule.
7. Place the lid on the strip and keep the strip horizontal to prevent the mineral oil from dripping.
8. Incubate the strip and the Urea-Arginine LYO 2 (MYCOPLASMA R1 + MYCOPLASMA R2) broth remaining in the
MYCOPLASMA R2 vial for 24 ± 4 hours and 48 ± 6 hours at +36°C ± 2°C in aerobic conditions.

RESULTS AND INTERPRETATION

Note:
For further details on reading and interpretation of results, refer to the reading guide at the end of this package insert.
Urea-Arginine LYO 2 (MYCOPLASMA R1 + MYCOPLASMA R2):
1. Read the Urea-Arginine LYO 2 broth color change after 24 hours. In case of a negative result, extend incubation to
48 hours and read again.

Positive Uninterpretable
Negative
(Ureaplasma spp. and/or M. hominis)
Turbid broth, whatever the color.
Broth color
Yellow Orange to red*
change Any color other than yellow or orange
to red.

 Some semen samples may produce a dark yellow color in the vials when reconstituting the R2 broth and when
inoculating the Uspp cupules of the strip. In such a case, a positive result should be interpreted as a color change from
this baseline shade.
Note:
If the Urea-Arginine LYO 2 (MYCOPLASMA R1 + MYCOPLASMA R2) broth is uninterpretable, do not read the strip.

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MYCOPLASMA IST 3 strip

Reading
1. Read the cupules after 24 ± 4 hours and 48 ± 6 hours.
2. Record the results obtained on the result sheet at the end of this package insert.
Interpretation
For identification and indicative enumeration, interpretation should be performed as soon as an identification is
obtained (after either 24 or 48 hours). In case of a mixed culture, each species may be identified at different times.
A negative result can only be interpreted after 48 hours.

For Antimicrobial Susceptibility Testing, interpretation should be performed for identified species after 24 hours if the
≥ 106 cupule is positive or otherwise after 48 hours.
Identification Enumeration Susceptibility tests for Uspp Identification Enumeration Susceptibility tests for M. hominis

Uspp Uspp Uspp LVX LVX MXF MXF TET TET ERY ERY TEL Mh Mh LVX LVX MXF MXF TET TET CLI CLI
0 Uspp Mh
≥103 ≥104 ≥106 2 4 2 4 1 2 8 16 4 ≥104 ≥106 1 2 0.25 0.5 4 8 0.25 0.5
Positive reading Orange/Orangey-red/Red/Dark red/Fuschia

Negative reading Yellow / Dark yellow

The strain is: The strain is:
Presence - - Susceptible (S) - - Susceptible (S)
Interpretation of of Uspp Presence Uspp Uspp Uspp a) Presence of Mh Mh b)
+ - Resistant (R)/Susceptible (S) + - Resistant (R)/Susceptible (S)
positivity and/or of Uspp ≥103 ≥104 ≥106 M. hominis ≥104 ≥106
M. hominis + + Resistant (R) + + Resistant (R)
- + Uninterpretable - + Uninterpretable

a) For Ureaplasma spp., the AST result of the + - profile should be interpreted as Resistant (R) for all antibiotics except Tetracycline in
the TET cupule which should be interpreted as Susceptible (S).
b) For Mycoplasma hominis, the AST result of the + - profile should be interpreted as Resistant (R) for all antibiotics except Clindamycin
in the CLI cupule which should be interpreted as Susceptible (S).

According to the CLSI M43-A guidelines (2), organisms susceptible to Tetracycline will also be susceptible to
Doxycycline, and organisms susceptible to Erythromycin will also be susceptible to Azithromycin.

Note 1:
Identification/enumeration and Antimicrobial Susceptibility Testing may be interpreted at different times for a given
species.
Note 2:
The results are uninterpretable if:
- The lower enumeration cupule is negative and a higher enumeration cupule is positive (for example  104 is
negative and  106 is positive).
- The growth control (0) is negative and some identification and enumeration cupules are randomly positive.
- The growth control (0) is positive and some other cupules are randomly positive.
- A result is negative at the lowest concentration of an antibiotic and positive at the highest concentration. In this
case, the test should be repeated.
Note 3:
If the titer is low, the color change may be observed in the Urea-Arginine LYO 2 vial only and not in the growth control on
the strip (the titer of the specimen is too low to produce the color change). The test should not be interpreted.

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QUALITY CONTROL
The strips and media are systematically quality controlled at various stages of their manufacture. For users who wish to
perform their own quality control tests with the kit, the following strains should be used:
1. Ureaplasma parvum ATCC® 27815TM
2. Ureaplasma urealyticum ATCC® 33175TM
3. Mycoplasma hominis ATCC® 23114TM
Protocol
1. Culture the strain in the Urea-Arginine LYO 2 broth (ref. 42508).
2. Transfer 30 μL of this vial into a new 3 mL Urea-Arginine LYO 2 vial (using an additional Urea Arginine LYO 2 kit,
Ref. 42508) and perform 3 successive 1:10 dilutions in Urea-Arginine LYO 2 vials (using an additional Urea Arginine
LYO 2 kit, Ref. 42508).
3. Incubate these 4 Urea-Arginine LYO 2 vials at +36°C ± 2°C until sufficient growth is obtained (about 16 to 24 hours of
incubation).
4. For Ureaplasma spp., transfer 15 µL of the most diluted Urea-Arginine LYO 2 vial that is positive into a new vial of
Urea-Arginine LYO 2 broth (vials included in the current MYCOPLASMA IST 3 kit, Ref. 422083).
For M. hominis, using the most diluted Urea-Arginine LYO 2 vial that is positive, perform a 1:10 dilution in NaCl
0.85% and transfer 15 μL of this dilution into a new vial of Urea-Arginine LYO 2 broth (vials included in the current
MYCOPLASMA IST 3 kit, Ref. 422083).
This inoculum will be used to inoculate the MYCOPLASMA IST 3 strip.
Expected results

Identifi-
Identification Enumeration Susceptibility tests for Ureaplasma spp.
Enumeration Susceptibility tests for M. hominis
cation
Reading Uspp Uspp Uspp LVX LVX MXF MXF TET TET ERY ERY TEL Mh Mh LVX LVX MXF MXF TET TET CLI CLI
0 Uspp Mh
time ≥103 ≥104 ≥106 2 4 2 4 1 2 8 16 4 ≥104 ≥106 1 2 0.25 0.5 4 8 0.25 0.5
1. U. parvum 24h + + + + V - - - - - - - - - - - - - - - - - - - -
ATCC® 27815TM 48h + + + + V - - - - - - - - - - - - - - - - - - - -
2. U. urealyticum 24h + + + + V - - - - + + - - - - - - - - - - - - - -
ATCC® 33175TM 48h + + + + V - - - - + + - - - - - - - - - - - - - -
3. M. hominis 24h V - - - - - - - - - - - - - V V V - - - - - - V -
ATCC® 23114TM 48h + - - - - - - - - - - - - - + + V - - - - - - V -

V = variable

Note:
If the  106 cupule is positive after 24 hours of incubation, perform an additional 1:10 dilution of the sample and repeat
the test.
It is the responsibility of the user to perform Quality Control taking into consideration the intended use of the medium,
and in accordance with any applicable local regulations (for example, frequency, number of strains, incubation
temperature).

LIMITATIONS OF THE METHOD

 A specimen cannot be considered as negative before 48 hours of incubation.
 If the specimen titer is low (positive Urea-Arginine LYO 2 broth), the strip cupules may not change color or the color
change may be inconsistent.
 Enumeration performed in the strip test cupules only provides an indication of the titer. The exact titer should be
determined based on culture methods using specific agars (CLSI M43).
 Some Ureaplasma spp. strains that are highly resistant to Erythromycin (MIC ≥ 128 µg/mL) may also grow in the Mh
part of the strip, leading to the interpretation of the presence of a Clindamycin-resistant Mycoplasma hominis strain.
Since this pattern is highly unlikely, the species present in the sample should be confirmed using an alternative
method.
 Some Mycoplasma hominis strains that are resistant to Lincomycin (MIC ≥ 8 µg/mL) may also grow in the Uspp part
of the strip, leading to the interpretation of the presence of a Clindamycin-resistant Ureaplasma spp. strain. Since this
pattern is unlikely, the species present in the sample should be confirmed using an alternative method.
 Use of compounds containing the active ingredients listed below may have an inhibitory effect on the results obtained
with the MYCOPLASMA IST 3 strip: condoms (latex), lubricant gel (aloe vera, vitamin E), titanium oxide, bisacodyl,
dodeclonium bromide, esculoside sesquihydrate, enoxolone, benzocaine and promestriene.
 The presence of human blood in the clinical samples could interfere with the results since human blood cells could
mask the coloration and make the color change reading difficult to perform. However, during product development,
no false results caused by the presence of human blood were observed.

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PERFORMANCE
MYCOPLASMA IST 3 performance was evaluated at three clinical laboratories in France, the United Kingdom and
Serbia. 516 clinical samples (vaginal/cervical swabs (n=230, 44.6%), urethral swabs (n=56, 10.9%), semen (n=88,
17.1%) and male urines (n=142, 27.5%) were prospectively included in this trial. A composite sample status was defined
based upon A7 agar (bioMérieux, Ref. 43003) and Polymerase Chain Reaction (PCR) results. The use of A7 agar also
allowed to estimate sample density. The antimicrobial susceptibility of the isolates was then tested using CLSI-M43A
compliant Broth Microdilution. From this population, 312 samples had a negative status and 204 (including 78 (38.2%)
contrived samples) had a positive status, including 22 mixed samples.
Detection and Identification:
MYCOPLASMA IST 3 identification results were compared to composite sample statuses. Positive and Negative
Agreements [95% Confidence Interval] were as follows:

Positive Agreement Negative Agreement

97.5 [94.4-99.2]% 99.7 [98.2-100.0]%

At sample levela (199/204) (311/312)

At Ureaplasma spp. 98.5 [94.6-99.8]% 99.7 [98.6-100.0]%

species level (129/131) (384/385)
At Mycoplasma hominis 99.0 [97.5-99.7]%
92.6 [85.6-96.4]%
species level
(88/95) (410/414b)

a
The sample level accounts exclusively for the sample status (i.e., a positive sample may be contaminated by
Ureaplasma spp. and/or Mycoplasma hominis).
b
Mycoplasma hominis status was not determined for 7 samples.
Note:
- During the clinical trials, 4 samples artificially contaminated with Erythromycin-resistant Ureaplasma spp., also
produced growth in the Mycoplasma hominis identification, enumeration and Clindamycin cupules.
- During the clinical trials, 22 samples contaminated with both Ureaplasma spp. and Mycoplasma hominis were
observed. MYCOPLASMA IST 3 allowed full recovery of both species for 18 samples and partial recovery of
Ureaplasma spp. only for the 4 remaining samples.
Indicative enumeration:
MYCOPLASMA IST 3 indicative enumeration results were compared to results obtained using A7 agar for samples with
positive concordant species with A7 agar (cut-off at 104 CFU/mL for both methods). One hundred and seventeen
samples contaminated with Ureaplasma spp. and 86 samples contaminated with Mycoplasma hominis, including 17
mixed samples, were analyzed. Performance was expressed as an exact agreement with A7 agar indicative enumeration
results, and as an agreement within ± 101 CFU/mL with A7 agar results, as both MYCOPLASMA IST 3 and A7 agar
provide an indication of sample density and not an exact titer. Results were as follows:

Within ± 101 CFU/mL

Exact agreement
agreement
86.0 [80.3-90.3]% 97.8 [94.6-99.4]%
Combined species*
(160/186) (182/186)

At Ureaplasma spp. 86.3 [78.9-91.4]% 100.0 [96.9-100.0]%

species level (101/117) (117/117)

At Mycoplasma hominis 83.7 [74.5-90.0]% 94.2 [87.1-97.5]%

species level (72/86) (81/86)
 For mixed samples, a concordant sample may be concordant for Ureaplasma spp. and/or Mycoplasma hominis.
Note:
The Uspp ≥ 103 CFU/mL enumeration cupule is relevant for the male urines. Among the 19 urines tested during the trial
with a positive status for Ureaplasma spp., all had a density ≥ 103 CFU/mL using A7 agar.
Using MYCOPLASMA IST 3, 17 samples also had a density ≥ 103 CFU/mL. The two remaining samples had a density of
103 CFU/mL using A7 agar and < 103 CFU/mL using MYCOPLASMA IST 3. Note that these two samples are a worst
case as the A7 agar interpretation does not allow a result <103 CFU/mL while growth may be variable for samples with a
density ≤ 102 CFU/mL.

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Antimicrobial Susceptibility Testing (AST):

Interpretive categories obtained with MYCOPLASMA IST 3 were compared to MICs obtained using the CLSI M43-A
Broth Microdilution reference method and related interpretive breakpoints for samples with positive concordant species
which have composite sample statuses. The AST performance is summarized in the following table (CA: Category
Agreement, ME: Major Error, VME: Very Major Error, R: Resistant, S: Susceptible):

Isolate status (BMD) Performance index

Ureaplasma spp. Total #S #R CA (%) ME (%) VME (#)
96.0% 4.2%
Levofloxacin 124 120 4 0
(119/124) (5/120)
98.4% 1.6%
Moxifloxacin 124 123 1 0
(122/124) (2/123)
97.6% 1.7%
Tetracycline 124 120 4 1
(121/124) (2/120)
99.2% 0.8%
Erythromycin 124 120 4 0
(123/124) (1/120)
99.2% 0.8%
Telithromycin 125 122 3* (124/125) (1/122)
0
98.1% 1.8%
All drugs combined 621 605 16 1
(609/621) (11/605)
Mycoplasma hominis Total #S #R CA (%) ME (%) VME (#)
98.8% 1.2%
Levofloxacin 84 82 2 0
(83/84) (1/82)
97.6% 1.2%
Moxifloxacin 84 81 3 1
(82/84) (1/81)
97.6% 1.5%
Tetracycline 84 65 19 1
(82/84) (1/65)
100.0% 0.0%
Clindamycin 84 84 0 N/A
(84/84) (0/84)
98.5% 1.0%
All drugs combined 336 312 24 2
(331/336) (3/312)
All drugs and species 98.2% 1.5%
957 917 40 3
combined (940/957) (14/917)
 No Resistant breakpoints are published for Telithromycin; these 3 isolates were found to be non-susceptible to
Telithromycin.
Note:
- For Ureaplasma spp.: among the 12 categorical errors observed during the trial, 8 out of the 11 MEs and the single
VME originated from isolates with MIC values within ± 1 doubling dilution of the MIC defining the CLSI interpretive
breakpoints, whereas no intermediate category exists for those antimicrobial agents. The remaining MEs observed
for:
o Erythromycin, originated from an isolate with an MIC of 0.5 µg/mL.
o Moxifloxacin, originated from an isolate with an MIC of 0.5 µg/mL.
o Telithromycin, originated from an isolate with an MIC of 0.125 µg/mL.
- For Mycoplasma hominis: among the 5 categorical errors observed during the trial, 1 out of the 3 MEs and the 2
VMEs originated from isolates with MIC values within ± 1 doubling dilution of the MIC defining the CLSI interpretive
breakpoints, whereas no intermediate category exists for those antimicrobial agents. The remaining 2 MEs observed
for:
o Levofloxacin, originated from an isolate with an MIC of 0.25 µg/mL.
o Tetracycline, originated from an isolate with an MIC of 0.5 µg/mL.

Interfering Substances
The interference of several substances with the MYCOPLASMA IST 3 test was studied. The substances were from
endogenous and exogenous sources: compounds introduced during patient treatment, contaminants inadvertently
introduced during specimen collection and metabolites produced in pathological conditions.
No interference was observed for ketoprofen, hydrocortisone, aciclovir, urea, glucose, zinc oxide, vaseline, glycerin, talc
and compounds from intimate hygiene wipes and tampons.
See the SPECIMEN COLLECTION AND PREPARATION and LIMITATIONS OF THE METHOD sections for a list of
interfering substances.

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IMPORTANT OBSERVATIONS:
As with all Antibiotic Susceptibility Testing data, MYCOPLASMA IST 3 results are in vitro values only and may provide an
indication of the organism’s potential in vivo susceptibility. The use of results to guide therapy selection must be the sole
decision and responsibility of the attending physician who should base judgement on the medical history and knowledge
of the patient, pharmacokinetics/pharmacodynamics of the antibiotic and clinical experience in treating infections caused
by the particular bacterial pathogen. The antibiotic, dose and dosing regimen must also be considered.
For details of specific interpretive limitations and/or limitations on the clinical use of an antibiotic in various therapeutic
situations, please refer to the tables and footnotes of MIC interpretive standards in the latest CLSI® AST documents for
dilution procedures (M43) (2).

WASTE DISPOSAL
Unused reagents may be considered as non-hazardous waste and disposed of accordingly.
Dispose of used reagents as well as any other contaminated disposable materials following procedures for infectious or
potentially infectious products.
It is the responsibility of each laboratory to handle waste and effluents produced, according to their nature and degree of
hazardousness, and to treat and dispose of them (or have them treated and disposed of) in accordance with any
applicable regulations.

LITERATURE REFERENCES
1. WAITES K.B., DUFFY L.B. et al.
Standardized methods and quality control limits for agar and broth microdilution susceptibility testing of Mycoplasma
pneumoniae, Mycoplasma hominis and Ureaplasma urealyticum - JCM, nov 2012, 50, 3542-3547.
2. Methods for antimicrobial susceptibility testing for human mycoplasmas; approved guideline - M43-A Vol 31,9.
3. BAURIAUD R., SEROR C., LARENG M.B. et al.
Sensibilité in vitro aux antibiotiques des mycoplasmes génitaux isolés à Toulouse. Etude de nouvelles molécules
(macrolides et quinolones) - Path. Biol., 1992, 40, 479-482.
4. BEBEAR C., BEBEAR C.M.
Infections humaines à mycoplasmes - Rev. Fr. Lab., 2007, 391, 63-69.
5. BEBEAR C.M., DE BARBEYRAC B., PEREYRE S., BEBEAR C.
Mycoplasmes et chlamydiae : sensibilité et résistance aux antibiotiques – Rev. Fr. Lab., 2007, 391, 77-85.
6. BEBEAR C., DE BARBEYRAC B., BERNET C. et al.
Méthodes d'exploration des infections à mycoplasmes - Ann. Biol. Clin.,1989, 47, 415-420.
7. BEBEAR C., DE BARBEYRAC B., DEWILDE A. et al.
Etude multicentrique de la sensibilité in vitro des mycoplasmes génitaux aux antibiotiques - Path. Biol., 1993, 41, 4,
289-293.
8. KENNY G.E., CARTWRIGHT F.D. - Effect of pH, Inoculum Size, and Incubation Time on the Susceptibility of
Ureaplasma urealyticum to Erythromycin In Vitro - Clin. Infect. Dis., 1993, 17, Suppl. 1, 215-218.
9. TAYLOR-ROBINSON D., BEBEAR C.
Antibiotic susceptibilities of mycoplasmas and treatment of mycoplasmal infections – J. Antimicrob. Chemother.,
1997, 40, 622-630.
10. WAITES K.B., BEBEAR C.M., ROBERTSON J.A., TALKINGTON D.F., KENNY G.E.
CUMITECH 34: Laboratory diagnosis of mycoplasmal infections – Ed American Society for Microbiology, 2001, 1-30.

bioMérieux SA English - 9
MYCOPLASMA IST 3 050923-02 - IVD - en - 2020-03

INDEX OF SYMBOLS

Symbol Meaning
Catalogue number

In Vitro Diagnostic Medical Device

Manufacturer

Temperature limit

Use by date

Batch code

Consult Instructions for Use

Contains sufficient for <n> tests

Do not re-use

Date of manufacture

LIMITED WARRANTY
bioMérieux warrants the performance of the product for its stated intended use provided that all procedures for usage,
storage and handling, shelf life (when applicable), and precautions are strictly followed as detailed in the instructions for
use (IFU).
Except as expressly set forth above, bioMérieux hereby disclaims all warranties, including any implied warranties of
merchantability and fitness for a particular purpose or use, and disclaims all liability, whether direct, indirect or
consequential, for any use of the reagent, software, instrument and disposables (the "System") other than as set forth in
the IFU.

REVISION HISTORY
Change type categories:
N/A Not applicable (First publication)
Correction Correction of documentation anomalies
Technical change Addition, revision and/or removal of information related to the product
Administrative Implementation of non-technical changes noticeable to the user
Note: Minor typographical, grammar, and formatting changes are not included in the
revision history

Release Part Number Change Type Change Summary

date
Interfering substances
2020/03 050923-02 Technical change Interpretation of the TEL test
Quality control

2019/10 050923-01 N/A First publication

BIOMERIEUX, the BIOMERIEUX logo, PolyVitex and ATB are used, pending and/or registered trademarks belonging to bioMérieux, or
one of its subsidiaries, or one of its companies.
CLSI is a trademark belonging to Clinical Laboratory and Standards Institute Inc.
The ATCC trademark and trade name and any and all ATCC catalog numbers are trademarks of the American Type Culture Collection.
Any other name or trademark is the property of its respective owner.

673 620 399 RCS LYON

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69280 Marcy-l'Etoile - France www.biomerieux.com
MYCOPLASMA IST 3 050923-02 - IVD - en - 2020-03

RESULT SHEET
MYCOPLASMA IST 3 Specimen:

Identifi-
Identification Enumeration Susceptibility tests for Ureaplasma spp.Enumeration Susceptibility tests for M. hominis
cation
Reading Uspp Uspp Uspp LVX LVX MXF MXF TET TET ERY ERY TEL Mh Mh LVX LVX MXF MXF TET TET CLI CLI
0 Uspp Mh
time ≥103 ≥104 ≥106 2 4 2 4 1 2 8 16 4 ≥104 ≥106 1 2 0.25 0.5 4 8 0.25 0.5
24h
48h

673 620 399 RCS LYON

bioMérieux SA Tel. 33 (0)4 78 87 20 00
376 Chemin de l'Orme Fax 33 (0)4 78 87 20 90
69280 Marcy-l'Etoile - France www.biomerieux.com
MYCOPLASMA IST 3 050923-02 - IVD - en - 2020/03

READING GUIDE
SAMPLE PREPARATION
Dry swabs (urethral or
INOCULATION
cervical/vaginal):
direct release of the 3 mL
sample in R1 TRANSPORT Mix using a
Liquid samples (semen, vortex-type
Storage: mixer until
urine or transport media
20h at +18°C/+25°C complete
of urethral or cervical/
vaginal ESwab®): 72h at +2°C/+8°C dissolution
200 µL in R1
R1 R1 R2 R1+R2

DISTRIBUTION MYCOPLASMA IST3

0 Uspp ≥103 ≥106

55 µL 2 drops

≥104
x x
25 cupules 25 cupules

LVX MXF TET ERY TEL

4
2 2 1

4
2
16
STEP STEP
8

1 2
4 Mh ≥104 1 0.25 4 0.25

R1+R2
≥106

Mineral oil
LVX MXF TET CLI

2
0,5
8
0,5

INCUBATION OF STRIP AND BROTH (R1 + R2) : 24 ± 4 hours and 48 ± 6 hours at +36 ± 2°C in aerobic conditions

R1 + R2 DETECTION After 24 ± 4 hours and 48 ± 6 hours

Negative (Yellow) Positive (Orange to red) Uninterpretable

Absence of Ureaplasma spp. Presence of Ureaplasma spp. Turbid broth, whatever the color.
and M. hominis and/or M. hominis Any color other than yellow or
orange to red

INTERPRETATION OF THE STRIP CUPULE COLORS

Color Interpretation

Yellow
Negative
Dark yellow*

Orange

Orangey-red

Red Positive

Dark red

Fuschia

* Some semen samples may produce a dark yellow color close to orange when inoculating the Uspp cupules.
In such a case, a positive result should be interpreted as a color change from this baseline shade.

bioMérieux SA English - Reading Guide - 1

MYCOPLASMA IST 3 050923-02 - IVD - en - 2020/03

READING AND INTERPRETATION After 24 ± 4 hours and 48 ± 6 hours

NEGATIVE GROWTH CONTROL UNINTERPRETABLE

and all other cupules negative

MYCOPLASMA IST3
MYCOPLASMA IST3

MYCOPLASMA IST3
MYCOPLASMA IST3
≥104 ≥106

0 0 0 Uspp ≥103 ≥106 Mh ≥104

0

Absence of Ureaplasma spp. If the growth control is

and M. hominis negative and at least one
other cupule is positive

Identification and Enumeration of Ureaplasma spp.

ABSENCE OF PRESENCE OF UREAPLASMA SPP.
UREAPLASMA SPP.
MYCOPLASMA IST3

MYCOPLASMA IST3

MYCOPLASMA IST3

MYCOPLASMA IST3

MYCOPLASMA IST3
≥104 ≥104 ≥104 ≥104 ≥104

0 Uspp ≥103 ≥106 0 Uspp ≥103 ≥106 0 Uspp ≥103 ≥106 0 Uspp ≥103 ≥106 0 Uspp ≥103 ≥106

<103 CFU/mL ≥103 CFU/mL ≥104 CFU/mL ≥106 CFU/mL

<104 CFU/mL <106 CFU/mL

Identification and Enumeration of Mycoplasma hominis

ABSENCE OF M. HOMINIS PRESENCE OF M. HOMINIS
MYCOPLASMA IST3

MYCOPLASMA IST3

MYCOPLASMA IST3

MYCOPLASMA IST3

≥106 ≥106 ≥106 ≥106

0 Mh ≥104 0 Mh ≥104 0 Mh ≥104 0 Mh ≥104

≥104 CFU/mL
<104 CFU/mL ≥106 CFU/mL
<106 CFU/mL

Susceptibility Test Reading

For the species identified, interpretation should be performed after 24 hours if the ≥ 106 cupule is positive or otherwise after
48 hours.

TESTS FOR UREAPLASMA SPP. TESTS FOR M. HOMINIS

4 4 2 16 2 0.5 8 0.5

2 2 1 8 4 1 0.25 4 0.25
LVX MXF TET ERY TEL LVX MXF TET CLI

All susceptibility tests except TEL TEL

Susceptible
S S S S S S S S
Non-susceptible
R R S R R R R S

R R R R R R R R

Uninterpretable Uninterpretable

bioMérieux SA English - Reading Guide - 2