ADIAVET MYCO AV

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ADIAVET MYCO AV
Commercial PCR kit for the detection of
Mycoplasma gallisepticum, Mycoplasma synoviae
and Mycoplasma meleagridis from tracheal swabs
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Products - ADIAVET MYCO AV

• Reagents contained in the kit

A1 G Reagent amplification solution 1 for
M.gallisepticum
E2 Reagent loading buffer
A1 SM Reagent amplification solution 1 for
M.synoviae and M.meleagridis
E1 Reagent size marker
A2 Reagent amplification solution 2
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Main objections for PCR diagnosis
Technical risks - Contamination by PCR product
inducing false positives Inhibition of PCR
reaction inducing false negatives Financial
objections - Royalties due to Hoffmann La Roche
for the use of PCR
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Solutions of the Adiavet range
Technical risks
Include negative sample to check the probleme of
contamination false positive control
An internal control avoiding false negatives.

• Financial objections
• Kits are sold under a Hoffmann La Roche licence.
  No more licence is owed
• PCR method is so sensitive that for some of our
  tests, there is a possibility to pool the
  samples, reducing the costs.

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Solutions of the Adiavet range
Other advantages
A coloured polymerase to confirm its presence in
the tube. A specific ladder in each kit
corresponding to the size of the amplified
product for the elctrophoresis
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ADIAVET MYCO AV
Multiplex PCR for M. synoviae and M.
meleagridis Single PCR for M. gallisepticum
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ADIAVET MYCO AV
Specificity
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ADIAVET MYCO AV

The sensitivity of each PCR test (MG, MS-MM) was
evaluated with serial dilution of positive
liquide culture. The detection level obtained on
pure culture are 0.5 CCU Mycoplasma
gallisepticum 0.5 CCU Mycoplasma synoviae 1 CCU
Mycoplasma meleagridis
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ADIAVET MYCO AV
The sensitivity of the PCR test allows pool

Individual PCR
Pool 3 by 3
RESULT - -

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ADIAVET MYCO AV

PCR Results Results are only qualitative
Negative result mycoplasma contamination
level is under the PCR detection level (0,1 to
1Unit Changing Color). Positive mycoplasma
contamination level is upper the PCR detection
level
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ADIAVET MYCO AV

Notes
If mycoplasma strains are needed (MIC
determination), swabs could be used for culture
before DNA extraction.
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ADIAVET MYCO AV

Field assessment
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ADIAVET MYCO AV

Mycoplasma synoviae PCR
94 swabs representing 6 flocks Turkey or
chichen breeders flocks showing respiratory
infection and decreased egg production Two of
them were regularly MS infected. Swabs were
treated individually or pooled 3 by 3. In each
case 5/6 flocks were MS PCR positive
(confirmed by RSA) 1/6 flock was MS PCR and
RSA negative
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ADIAVET MYCO AV

Mycoplasma gallisepticum PCR
46 swabs representing 4 flocks Turkey and poultry
flocks showing infectious sinusitis, coughing,
mortality 2/4 flocks were MG PCR positive
(confirmed by RSA and culture) 2/4 flocks were
MG PCR negative (confirmed by RSA and culture).
These two flocks were under antibiotic treatment.
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ADIAVET MYCO AV

Mycoplasma meleagridis PCR
1 turkeys breader known to be contaminated by
Mycoplasma meleagridis 172 swabs. 35 were MM
PCR positive 36 were MM culture positive
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ADIAVET MYCO AV

Other breeding farms
1 flock without mycoplasma problem over the last
years 30 swabs MG/MS/MM PCR test No PCR positif
sample 1 flock without any clinical sign 11
swabs. MG/MS/MM PCR test No PCR positif sample
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ADIAVET MYCO AV
Field validation of the test
Method Clinical signs and Culture / Serology
Method PCR
318 samples from 15 flocks were tested.
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ADIAVET MYCO AV
PCR vaccination

• -The PCR target is a constitutive gene present on
  all mycoplasma strains including vaccine strain
• No information about the vaccine strain
  (mutation, deletion.)

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ADIAVET MYCO AV
PCR vaccination

• After vaccination, vaccine strain persist during
  2 to 3 weeks
• vaccination doesnt induce serological response
• PCR and ELISA Infection

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ADIAVET MYCO AV
Conclusion

Our two PCR tests are very specific (detect
only MG-MS-MM) very sensitive very rapid
tightly related to clinical signs and other
diagnostic tests (culture,ELISA,RSA)
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