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Updated: Jan 3, 2026
Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
Published on: August 29, 2014
Use of next-generation amplicon sequencing to study Blastocystis genetic diversity in a rural human population from
Liliana Rojas-Velázquez1,2, Jenny G Maloney3, Aleksey Molokin3
1Unidad de Investigación en Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), Mexico City, Mexico.
Background:
The intestinal parasite Blastocystis is found in humans and animals around the world. It is spread through the consumption of contaminated food and water and has been associated with a variety of intestinal symptoms. Blastocystis is one of the most common intestinal parasites in humans, yet its prevalence and distribution in humans in North America is not well characterized.
Methods:
Next-generation amplicon sequencing of a region of the Blastocystis SSU rRNA gene was applied to DNA extracted from fecal specimens obtained from 182 inhabitants of a rural population in Mexico to characterize Blastocystis prevalence, subtype distribution, and intra-host subtype diversity in humans.
Results:
Of the 182 samples tested in this study, 68.1% (124) contained one or more Blastocystis subtypes. Subtype 3 was the most common subtype observed and was found in 81.5% of the positive samples. Subtype 1, 16.9% of the positive samples, and subtype 2, 17.7% of the positive samples, were also found in this population. Mixed infections were observed in 13.7% of the positive samples. In this population, the odds of having Blastocystis increased in adulthood (> 15 years; OR: 1.72, P < 0.0001), and the odds of having subtype 1 increased in the presence of farm animals (OR: 1.51, P = 0.03). The odds of having subtype 1, subtype 2, or a mixed infection decreased in the presence of cement flooring (OR: - 1.61, P = 0.005; OR: - 1.14, P = 0.03; OR: - 1.48, P = 0.02) possibly indicating socioeconomic factors are involved in the risk of acquiring one of these subtypes.
Conclusions:
These data contribute to our understanding of the epidemiology of Blastocystis infection in humans and can be used to shape future studies which aim to better characterize the transmission pathways and health outcomes of Blastocystis infections.
Insights
Blastocystis, a common intestinal parasite, infects over 68% of a rural Mexican population, with subtype 3 being most prevalent. Factors like age and farm animal presence influence infection risk.
Area of Science:
- Microbiology
- Parasitology
- Epidemiology
Background:
- Blastocystis is a globally prevalent intestinal parasite transmitted via contaminated food and water.
- While common, its epidemiology in North America remains poorly understood.
- Associated with various gastrointestinal symptoms, its prevalence and subtype distribution require further characterization.
Purpose of the Study:
- To investigate the prevalence and distribution of Blastocystis subtypes in a rural Mexican population.
- To explore intra-host Blastocystis subtype diversity.
- To identify factors associated with Blastocystis infection and specific subtype acquisition.
Main Methods:
- Next-generation amplicon sequencing of the Blastocystis SSU rRNA gene.
- Analysis of DNA from fecal specimens of 182 human inhabitants.
- Statistical analysis to determine odds ratios for infection and subtype prevalence.
Main Results:
- Blastocystis was detected in 68.1% of individuals, with subtype 3 being the most common (81.5% of positive samples).
- Subtypes 1 and 2 were also identified, along with mixed infections in 13.7% of positive cases.
- Infection odds increased with age (>15 years), while presence of farm animals correlated with subtype 1. Cement flooring was associated with decreased odds of infection with subtypes 1, 2, or mixed infections, suggesting socioeconomic influences.
Conclusions:
- This study enhances understanding of Blastocystis epidemiology in humans.
- Findings provide a basis for future research on transmission pathways.
- Further investigation into health outcomes associated with Blastocystis infections is warranted.
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