PCR Detection and Molecular Identification of Chlamydiaceae Species

<jats:title>ABSTRACT</jats:title> <jats:p> Recent taxonomic developments, based on 16s and 23s rRNA gene sequences, have divided the family <jats:italic>Chlamydiaceae</jats:italic> into two genera and nine species, of which five have been found to infect humans. Few simple methods are available to detect and identify all species sensitively and specifically. In this study the suitability of the <jats:italic>omp2</jats:italic> gene as a target for molecular identification of <jats:italic>Chlamydiaceae</jats:italic> is demonstrated. Phylogenetic analysis of partial <jats:italic>omp2</jats:italic> gene sequences from all nine species agrees with the recently published taxonomic changes based on the ribosomal genes. The use of a family-specific PCR primer pair, which is able to amplify the 5′ end of the <jats:italic>omp2</jats:italic> gene from all <jats:italic>Chlamydiaceae</jats:italic> except some <jats:italic>Chlamydophila pecorum</jats:italic> strains, is described. Identification of all nine species was achieved using restriction fragment length polymorphism analysis with a single enzyme, <jats:italic>Alu</jats:italic> I, confirmed by DNA sequencing. A PCR enzyme-linked oligonucleotide assay was developed which can detect a single chlamydial genome and may be applied to DNA extracts from any specimen or culture for the detection of single or mixed human chlamydial infection. </jats:p>