1887

Abstract

A culture-independent genome sequencing approach was developed and used to examine genomic variability in -positive specimens that were collected from patients in the Seattle, WA, USA, area. The procedure is based on an immunomagnetic separation approach with chlamydial LPS-specific mAbs, followed by DNA purification and total DNA amplification, and subsequent Illumina-based sequence analysis. Quality of genome sequencing was independent of the total number of inclusion-forming units determined for the sample and the amount of non-chlamydial DNA in the Illumina libraries. A geographically and temporally linked clade of isolates was identified with evidence of several different regions of recombination and variable sequence types, suggesting that recombination is common within outbreaks. Culture-independent sequence analysis revealed a linkage pattern at two nucleotide positions that was unique to the genomes of isolates from patients, but not in recombinants generated . These data demonstrated that culture-independent sequence analysis can be used to rapidly and inexpensively collect genome data from patients infected by , and that this approach can be used to examine genomic variation within this species.

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2013-10-01
2022-01-19
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