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Title: IDENTIFICATION OF NINE SPECIES OF THE CHLAMYDIACEAE USING PCR-RFLP

Author
item EVERETT, KARIN
item Andersen, Arthur

Submitted to: International Journal of Systematic and Evolutionary Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/28/1998
Publication Date: N/A
Citation: N/A

Interpretive Summary: Species of Chlamydia are prevalent in animals ranging from pigs and poultry to humans and household cats. Diseases caused by these many bacteria can be severe, and chlamydiae are so diverse in virulence that identification is essential for proper managment and treatment. Some chlamydial pathogens even live in amoebae. Recently, all of the known chlamydial strains were classified into new groups and species. Of the existing methodologies tha might be used for chlamydial identification, only DNA sequencing has been developed for identifying the new groups. A test that would easily distinguish the new groupings has been needed. In this study a simple, rapid, and systematic PCR-RFLP procedure was developed by which laboratory cultured chlamydial specimens could be sorted into species This test provides a precise and easy method for identifying Chlamydia in livestock and other animals.

Technical Abstract: The family Chlamydiaceae contains two genera and nine species. Rapid and easy identification of these species is essential for taxonomic, epidemiologic, and clinical determinations. Currently, DNA sequence analysis is the only systematic method that distinguishes all nine species. In this study a simple, rapid, and systematic PCR-RFLP procedure was developed by which laboratory-cultured chlamydial specimens could be speciated. To accomplish this, conserved oligonucleotide primers and restriction sites were deduced from ribosomal sequence data from >50 chlamydial strains representing all nine species. DNA from 25 previously characterized chlamydial strains were tested with these primers and restriction enzymes. All nine chlamydial species were reliably distinguished in the tests. The procedure was optimized by adjusting the annealing temperature and using a modified DNA polymerase (AmpliTaq Gold) to reduce mismatch PCR amplification of mycoplasmas and other bacteria. Using the enzymes, primers, and methods described in this study, PCR-RFLP rapidly identifies known Chlamydiaceae species. NOTE: Patent potential to distinguish species - this paper related to patent application #0211.97 filed for invention entitled "Intergenic Spacer Target Sequence for Detecting and Distinguishiing Chlamydial Strains."