Use real-time PCR (qPCR) with primers specific to Bordetella pertussis DNA directly on the respiratory swab

Perform a traditional culture on the respiratory swab and use the isolated colonies for end-point PCR with specific primers

Start a traditional culture on the respiratory swab and then use the collected colonies for real-time PCR (qPCR)

Apply end-point PCR with specific primers for Bordetella pertussis DNA directly to the respiratory swab

The denaturation temperature — it ensures uniform melting of DNA strands

The extension time — it controls DNA synthesis efficiency

The primers — they provide specificity by binding to the target DNA sequence

The buffer composition — it maintains optimal pH during the reaction

It indicates whether the target DNA was present in the sample

It reveals the thermal cycling conditions used during PCR

It indicates the concentration of DNA in the original sample

It identifies the primer sequences used in the reaction

To monitor DNA amplification in real-time

To visualise PCR products using gel electrophoresis

To monitor temperature accuracy during thermal cycling

To increase the speed of the PCR reaction

By assessing the number of primers used in the qPCR reaction

By counting the total number of thermal cycles completed during the reaction

By assessing the cycle threshold (Ct) value

By interpreting the shape of the amplification curve to estimate the starting DNA concentration

It enables simultaneous detection of multiple pathogens

It lowers the risk of false-positive results

It increases the precision of pathogen identification

It reduces the number of thermal cycles required for amplification

To initiate amplification of the target DNA sequence

To allow primers to bind to the template DNA

To separate the DNA strands

To synthesise new DNA strands

The 16S rRNA gene is too conserved to be useful for bacterial identification

The PCR result suggests the bacterium is likely an environmental contaminant

The PCR product must be sequenced and analysed using a tool such as Nucleotide BLAST to identify the bacterium

The PCR product confirms the identity of the bacterium as Streptococcus pneumoniae