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Collection, Isolation, and Flow Cytometric Analysis of Human Endocervical Samples

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Analysis of multitag pyrosequence data from human cervical lavage samples.

Ammar Naqvi¹, Huzefa Rangwala, Greg Spear

¹Microbiome Analysis Center, Manassas, VA 20110, USA.

Chemistry & Biodiversity

|May 22, 2010

Summary

Researchers developed a new analysis pipeline to identify bacteria and fungi in human microbiome samples. This tool helps understand how microbial communities impact health, demonstrated using cervical vaginal lavage data.

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Multiplex Detection of Bacteria in Complex Clinical and Environmental Samples using Oligonucleotide-coupled Fluorescent Microspheres

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Published on: October 23, 2011

Area of Science:

Microbiology
Bioinformatics
Genomics

Background:

Human microbiome and mycobiome research generates vast sequencing data.
Systematic analysis is required to identify microbial taxa and correlate with clinical data.
Existing methods may lack efficiency for large-scale microbiome studies.

Purpose of the Study:

To develop and validate an automated analysis pipeline for microbiome and mycobiome data.
To enable rapid identification of microbial taxa and their correlation with host factors.
To apply the pipeline to Cervical Vaginal Lavage (CVL) samples for specific insights.

Main Methods:

Utilized Roche GS-FLX sequencing for bacterial and fungal community analysis.
Developed automated tools for data tracking, taxonomic analysis, and feature clustering.
Applied the pipeline to diverse sample types including stool, gut mucosa, vaginal, and oral washes.

Main Results:

Successfully generated tens of thousands of sequencing reads from various human body sites.
Demonstrated the pipeline's capability in analyzing CVL samples for microbial composition.
Established a framework for correlating microbiome data with clinical and environmental features.

Conclusions:

The developed analysis pipeline offers a systematic and rapid approach for microbiome research.
This tool is applicable to diverse microbiome and mycobiome datasets beyond CVL samples.
The pipeline will enhance understanding of the microbiome's influence on human health.