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Published on: March 29, 2017
Direct detection of Blastocystis sp. in human faecal samples and subtype assignment using single strand
Panagiotis G Menounos1, Gregory Spanakos, Nicholas Tegos
1Laboratory of Research, Nursing Military Academy, Athens, Greece.
Molecular and Cellular Probes
|August 3, 2007
Summary
This study developed a sensitive and specific polymerase chain reaction (PCR) assay to detect Blastocystis parasites in human stool. The method also subtypes the parasite, identifying subtype 3 as most common in Greece.
Area of Science:
- Medical Parasitology
- Molecular Biology
- Clinical Microbiology
Background:
- Blastocystis is an anaerobic intestinal parasite found in humans and other vertebrates.
- Classification into nine subtypes is based on small subunit ribosomal RNA (SSU rRNA) gene sequencing.
- The pathogenic role of Blastocystis and its various subtypes remains unclear.
Purpose of the Study:
- To develop a polymerase chain reaction (PCR)-based assay for direct detection of Blastocystis in human faeces.
- To enable subtype classification of Blastocystis using single-strand conformational polymorphism (SSCP) and sequencing.
- To facilitate large-scale epidemiological studies on Blastocystis prevalence and pathogenicity.
Main Methods:
- Development of a sensitive and specific PCR assay for Blastocystis detection in faecal samples.
- Integration of SSCP analysis and/or sequencing for parasite genotyping.
- Application of the protocol to 45 positive and 30 negative human faecal samples.
Main Results:
- The developed PCR assay demonstrated high sensitivity and specificity for Blastocystis detection.
- Genotyping revealed subtype 3 as the predominant Blastocystis subtype in Greece.
- Subtypes 1, 2, 4, 6, and 7 were also identified at lower frequencies.
Conclusions:
- A simple and effective PCR-based method for Blastocystis detection and subtyping has been established.
- The findings provide insights into the epidemiological distribution of Blastocystis subtypes in Greece.
- This method can aid future research into the clinical significance of Blastocystis infections.