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The mutation-specific antibody enables robust detection of the mutation in routine biopsy samples.
Using routine biopsy samples we successfully performed whole genomic microarray analysis to identify discriminative signatures.
Due to the instability of RNA and the suboptimal preservation of routine biopsy samples using formalin-fixed paraffin-embedded (FFPE) analyses, the methods discussed thus far for HPV transcript detection rely on the analysis of fresh-frozen tissue in research laboratories, therefore hampering the translation to routine clinical diagnostic.
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However, typically 10 routine biopsies can sample approximately only 0.2% of the prostate volume and therefore may not be representative of the entire cancer morphology panorama, even though the cancer detection rate may be 30 40%.
Tissue specimens investigated in the UK involved the collection of additional biopsy samples, during routine surgical investigations, with full written consent of the patient.
We propose several future lines of study that would be useful for providing more definitive answers in the event of a future seal die-off: Collection of blood samples from healthy seals to obtain baseline blood profiles as well as obtaining biopsy samples on a routine basis from dead animals.
Four patients (11%) had more advanced histologic signs in the biopsy samples, as identified during routine clinical examination.
Biopsy samples were processed for routine histology and for immunostaining with the lysosomal marker LAMP2 and the autophagosomal marker LC3.
The biopsy samples were processed using routine UCLH procedures – immediate fixation in standard formal saline at ambient temperature and same-day transfer to the laboratory for machine-automated processing, starting with formal saline and passing through standard alcohols and xylene to paraffin wax embedding, using the machine timings for biopsies.
This study was a retrospective one, with a molecular biology analysis carried out on biopsy samples performed according to the routine handling of such patients and families in our department.
Serial sections (10 μm) of the muscle biopsy samples were cut in a cryostat (−20 °C) and routine ATPase histochemistry analysis was performed after preincubation at pH 4.37, 4.60 and 10.30, as previously described (Andersen and Aagaard, 2000).
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