mRNA Detection
Active HPV Infection: E6/E7 mRNA Detection by Flow Cytometry
Introduction
A DNA test shows that HPV is present. It does not show whether the infection is doing anything: most infections are transient and clear on their own, and the ones that matter are those in which the viral oncogenes E6 and E7 are actively transcribed. Their mRNA is therefore a more direct readout of viral activity than viral DNA.
Reading that mRNA cell by cell is the harder part. The FLOWSCRIPT assay from Enzo Life Sciences uses oligonucleotide probes carrying a fluorophore and a quencher, so a signal appears only once a probe has hybridized to its target inside the intact cell. What is left is a fluorescence measurement any flow cytometer can take — with the single-cell resolution that a bulk PCR gives up.
The measurements below were made on the Agilent NovoCyte, the instrument family we supply and support. Ask us for a demonstration if you would like to see the assay run on your own samples. The application note is marked for research use only and not for use in diagnostic procedures.
Detection of HPV E6/E7 mRNA provides a direct indication of viral activity, offering greater clinical relevance than DNA testing alone. Using the FLOWSCRIPT HPV E6/E7 assay and the Agilent NovoCyte flow cytometer, researchers demonstrated accurate detection of these oncogene transcripts in both control cell lines and cervical cytology samples.
Key Findings
High accuracy in cell lines: HPV+ HeLa cells showed >95% positivity, while HPV– Jurkat cells remained <1%.
Reliable detection in clinical samples:All HPV+ cervical samples exceeded the 2% positivity threshold, while HPV– samples stayed below it.
Streamlined workflow: Wide dynamic range eliminates PMT adjustments, and optional automation with the NovoSampler Pro minimizes hands-on time.
Figure 2. E6/E7 mRNA expression detection in known negative and positive ectocervical cells. Cytology samples were evaluated for the presence of E6/E7 mRNA. Representative positive and negative samples are shown. Threshold is 2% cells within the E6/E7 gate. All HPV negative samples were <2% within the E6/E7 analysis gate, and all HPV positive samples were >2% of the E6/E7 gate.
Conclusion
The NovoCyte flow cytometer enables fast, reliable, and user-friendly single-cell detection of HPV E6/E7 mRNA, improving accuracy and efficiency in HPV research.
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