Evaluating HPV16 Gene Copy Number Measurement by SYBR Green PCR
Author Information
Author(s): Ian Roberts, Grace Ng, Nicola Foster, Margaret Stanley, Michael T Herdman, Mark R Pett, Andew Teschendorff, Nicholas Coleman
Primary Institution: MRC Cancer Cell Unit, Hutchison/MRC Research Centre, Cambridge, UK
Hypothesis
What are the errors associated with quantifying HPV16 gene copy numbers using SYBR green qPCR?
Conclusion
The study found that SYBR green qPCR produces significant inaccuracies in quantifying HPV gene copy numbers, particularly at low DNA concentrations.
Supporting Evidence
- Errors in quantifying HPV16 gene copy numbers ranged from 5% to 40%.
- Viral loads determined by qPCR showed reasonable agreement with Southern blotting.
- The E2/E6 ratio was found to be of limited value in assessing HPV physical state.
Takeaway
This study looked at how well a test measures the amount of a virus in samples from cervical cancer. It found that the test can make big mistakes, especially when there's not much virus.
Methodology
The study used SYBR green qPCR to measure HPV16 gene copy numbers in cervical cancer samples and compared results with Southern blotting.
Limitations
The study noted that there is unavoidable run-to-run variation in qPCR results, which can lead to significant errors in quantification.
Participant Demographics
The study involved cervical squamous cell carcinoma samples from women.
Digital Object Identifier (DOI)
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