Bio-Rad VINEO™ Brettanomytest PCR Kit User Manual

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You must use the the Opticon Monitor™ software (version 3.3 and higher)
or the CFX Manager™ software Industrial Diagnostic Edition for an
automated and quantitative interpretation. A complete report may be printed
(cf. software instructions).

1. Controls
Before a final interpretation of the results, you have to check the results of the
negative and positive controls.

For the test to be valid, the results of the negative and positive controls must
be as follows:

* N/A means “Not Applicable”. The software indicates N/A for the Ct of a sample when the
fluorescence curve does not cross the threshold.

If the results of the positive and negative PCR Qs controls are different from
those described in the table below, you have to begin the PCR again.

2. Samples
A sample is considered to be positive for Brettanomyces bruxellensis if the
value of Ct ≥ 10 is obtained for the FAM fluorophore.

If no value is obtained for Ct

FAM

(Ct

FAM

= N/A), the interpretation of the result

depends on the value of the internal control:

- A sample is considered to be negative for Brettanomyces bruxellensis if

no value of Ct is obtained for the FAM fluorophore (Ct

FAM

= N/A and if the

Ct of the internal control is greater than or equal to 28. (Ct

HEX

≥ 28).

- An N/A value for the Ct

HEX

of the internal control indicates that an

inhibition phenomenon of the PCR reaction has probably occurred, if
the Ct

FAM

of the target is also N/A. In this case, the DNA sample must

be diluted (to 1/10

for example) in sterile distilled water and then

subjected to a new PCR.

- If the Ct

HEX

of the internal control is less than 28, it is impossible to

interpret the result. Check that the threshold was set correctly or that
the raw curve shows an aspect characteristic of exponential
amplification. If the observed curve is not correct, it will be necessary to
repeat the PCR test for this sample.

Detection of Brettanomyces

bruxellensis (FAM)

Detection of the internal

control (Channel HEX)

Negative control

Ct = N/A*

25 ≤ Ct ≤ 40

Positive Qs PCR control

27 ≤ Ct ≤ 37

Not significant