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qPCR Is a Sensitive and Rapid Method for Detection of Cytomegaloviral DNA in Formalin-fixed, Paraffin-embedded Biopsy Tissue
Published on: July 9, 2014
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Cytopathology smears from autopsies: A viable storage method for molecular analysis
Lucas Hoelz1, Arya Mesgary2, Eduardo Achar1
1Santa Casa Medical School, São Paulo, Brazil.
Summary
Archived autopsy cytology smears provide high-quality DNA for molecular analysis. This method yields better DNA concentration and successful amplification compared to traditional histological specimens.
Area of Science:
- Forensic Science
- Molecular Biology
- Pathology
Background:
- Cytological smears are a viable alternative to histological samples for preserving autopsy material for DNA analysis.
- Archived air-dried smears from autopsies were evaluated for their utility in molecular analysis.
Purpose of the Study:
- To assess the feasibility of using archived air-dried cytological smears for DNA extraction and analysis.
- To compare the quantity and quality of DNA extracted from cytological smears versus corresponding histological specimens.
Main Methods:
- Air-dried cytological smears were collected from the same autopsy tissue areas designated for histology.
- DNA was extracted using a commercial kit, and its purity and concentration were determined.
- DNA integrity was assessed via conventional polymerase chain reaction (PCR).
Main Results:
- Five lung tumor cases (2 small cell carcinomas, 3 adenocarcinomas) were analyzed.
- Cytological smears yielded a median purity ratio of 1.87 and a mean DNA concentration of 2653 ng/mL.
- DNA from all cytological samples was successfully amplified by PCR, unlike any histological specimens.
Conclusions:
- Archived air-dried autopsy smears provide good quality and concentration of DNA suitable for conventional PCR.
- This cytological approach offers superior results compared to formalin-fixed paraffin-embedded histological material for DNA analysis.
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