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Updated: Dec 18, 2025
qPCR Is a Sensitive and Rapid Method for Detection of Cytomegaloviral DNA in Formalin-fixed, Paraffin-embedded Biopsy Tissue
Published on: July 9, 2014
Utility of two DNA extraction methods using formalin-fixed paraffin-embedded tissues in identifying congenital
Areli de la Cruz-de la Cruz1, Elsa Romelia Moreno-Verduzco2, Omar Martínez-Alarcón1
1Departamento de Infectología e Inmunología, Instituto Nacional de Perinatología, México, México.
Abstract:
Formalin-fixed paraffin-embedded (FFPE) tissues are a source of biological material for molecular studies; several methods to extract DNA from FFPE tissues have been reported. This process is challenging because of formaldehyde-induced cross-linking between proteins and DNA as well as molecule fragmentation when unbuffered formalin is used for fixation. Here, 2 methods for DNA extraction from FFPE tissues, based on a chelating resin and silica membrane columns, were modified and compared in their capacity to detect human cytomegalovirus (HCMV) in congenital infections. Both methods were tested on 121 samples of brain, lung, spleen, and liver derived from 36 deceased preterm newborns. Twelve patients were selected, and UL55 and UL75 HCMV genes were detected by polymerase chain reaction in 16/36 samples. These 2 methods represent a useful tool for DNA recovery from FFPE tissues and HCMV molecular identification with the advantage of low cost, minimal steps, minimal sample use, being solvent-free, and being easy to implement in the laboratory.
Insights
Two DNA extraction methods from formalin-fixed paraffin-embedded (FFPE) tissues were optimized for detecting human cytomegalovirus (HCMV) in congenital infections. These cost-effective, efficient techniques facilitate molecular identification in challenging FFPE samples.
Area of Science:
- Molecular Biology
- Virology
- Pathology
Background:
- Formalin-fixed paraffin-embedded (FFPE) tissues are valuable for molecular studies but challenging for DNA extraction due to formaldehyde-induced cross-linking and fragmentation.
- Existing DNA extraction methods require optimization for effective recovery from FFPE tissues, especially for detecting viral pathogens.
Purpose of the Study:
- To modify and compare two DNA extraction methods (chelating resin and silica membrane columns) for their efficacy in detecting human cytomegalovirus (HCMV) in FFPE tissues.
- To assess the utility of these methods for molecular identification of HCMV in congenital infections.
Main Methods:
- Modification and comparison of two DNA extraction techniques from FFPE tissues.
- Testing on 121 FFPE samples (brain, lung, spleen, liver) from 36 deceased preterm newborns.
- Detection of UL55 and UL75 HCMV genes using polymerase chain reaction (PCR).
Main Results:
- Both modified methods demonstrated effectiveness in DNA recovery from FFPE tissues.
- HCMV DNA was detected in 16 out of 36 samples (44.4%) using PCR.
- The methods were successful in identifying HCMV in congenital infections.
Conclusions:
- The modified chelating resin and silica membrane column methods are valuable tools for DNA recovery from FFPE tissues.
- These methods enable efficient molecular identification of HCMV, crucial for diagnosing congenital infections.
- Advantages include low cost, minimal steps, minimal sample usage, solvent-free operation, and ease of laboratory implementation.
