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Vivalytic Bordetella
– Instructions for Use
Reagents
All reagents necessary for the sample processing are integrated into the
cartridge. The processing includes cell lysis, nucleic acid extraction, DNA
amplification and detection.
Reagents are PCR bead, binding buffer, washing buffer, and elution buffer.
The PCR bead contains the DNA polymerase, primers, and probes. Binding
buffer facilitates binding of nucleic acids during the purification process.
Washing buffer is a formulation of different salts and solvents to remove
impurities e. g. proteins during the extraction process. Elution Buffer is
a low-salt buffer and contains the purified nucleic acids at the end of the
extraction process.
Sample Types / Medium
The test is intended to detect nucleic acids of B. pertussis (gene target
sequence IS481), B. parapertussis (gene target sequence pIS1001) and B.
holmesii (gene target sequences IS481, hIS1001) of nasopharyngeal swab
samples (flexible Flocked Swab FLOQSwabs® 553C) in eNAT® transport
medium (eNAT® transport medium Ref. 608C, COPAN Italia s.p.a.).
Collect and store samples as indicated in the manufacturer's instructions [3].
In case the sample is not processed immediately after sample collection,
nucleic acids will be preserved in eNAT® transport medium for up to 4 weeks
at room temperature and at 4°C and up to 6 months at 4 °C and up to 6
months at – 20 °C to – 80 °C (see Instructions for Use eNAT® transport
medium [3]).
Sample Preparation
Use a COPAN FLOQSwabs® flexible swab for collection of nasopharyngeal
swab samples. Each swab is transferred into 1 ml eNAT® transport medium.
Break the swab and close the tube. Shake the sample tube containing the
swab sample in eNAT® medium for homogenization (avoid foaming). Fill
300 μl of homogenized sample in the sample input of the cartridge and close
the cartridge lid. Insert the cartridge into the Vivalytic one analyser.
Do not use viscous samples that are difficult to pipette.
Test Result
After automatic processing of the sample with the Vivalytic one analyser the
test result is shown on the screen of the Vivalytic one analyser. The time to
result is about 50 min. For high titer specimens results are available after
less than 33 min and the test run can be terminated earlier (see chapter Test
Termination).
The sample is classified either as B. pertussis positive, B. parapertussis
positive, B. holmesii positive, negative or invalid or as a combination of the
aforementioned. In case of a positive detection of one or more targets the
test is considered valid even if the Human Control is negative.
Detection of the human cell based whole process control (Human Control)
in negative samples shows a successful DNA purification and amplification
procedure and excludes an inhibition of the PCR reaction. Interpretation of
results is shown in the table below.
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