PCR Detection and Molecular Identification of Chlamydiaceae Species
| Autor: | G. Ridgway, S. Kaye, J. C. Hartley, S. Stevenson, J. Bennett |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Microbiology (medical)
Chlamydiology and Rickettsiology Sequence analysis Chlamydiaceae Molecular Sequence Data Polymerase Chain Reaction Sensitivity and Specificity DNA sequencing law.invention Species Specificity law 23S ribosomal RNA Animals Cluster Analysis Humans Deoxyribonucleases Type II Site-Specific Phylogeny Polymerase chain reaction DNA Primers Genetics Chlamydophila Base Sequence biology Sequence Analysis DNA Ribosomal RNA biology.organism_classification Restriction fragment length polymorphism Polymorphism Restriction Fragment Length Bacterial Outer Membrane Proteins |
| Zdroj: | Journal of Clinical Microbiology. 39:3072-3079 |
| ISSN: | 1098-660X 0095-1137 |
| Popis: | Recent taxonomic developments, based on 16s and 23s rRNA gene sequences, have divided the family Chlamydiaceae 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 omp2 gene as a target for molecular identification of Chlamydiaceae is demonstrated. Phylogenetic analysis of partial omp2 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 omp2 gene from all Chlamydiaceae except some Chlamydophila pecorum strains, is described. Identification of all nine species was achieved using restriction fragment length polymorphism analysis with a single enzyme, Alu 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. |
| Databáze: | OpenAIRE |
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