PCR-test system specific identification Parachlamydia acanthamoebae

Abstract

Chlamydia-like bacteria Parachlamydia acanthamoebae belongs to the phylum Chlamydiae, class Chlamydiia, order Chlamydiales, family Parahlamydiaceae, genus Parachlamydia. The following terms were introduced to designate Chlamydiales bacteria that are not members of the Chlamydia genus: Chlamydia-related bacteria – CRB, Chlamydia-like bacteria and Chlamydia-like organisms – CLOs. Parachlamydia acanthamoebae is associated with respiratory diseases in humans and ruminants, as well as diseases of the reproductive organs leading to abortions, or the birth of weak or non-viable offspring. Unfortunately, there are no such test systems in the arsenal of humane and veterinary medicine of Ukraine, which could identify and differentiate Сhlamydia-related organisms, Parachlamydia acanthamoebae in particular. In this regard, PCR test system has been developed for the identification and species differentiation of Parachlamydia acanthamoebae, the bacteria of the Parachlamydiaceae family, which is the causative agent of chlamydiosis in ruminants and humans. Conducted bioinformatics studies revealed species-specific regions of the Parachlamydia acanthamoebae genome, which were used to develop the nucleotide structure of oligonucleotide primers, which allow to amplify the DNA fragment of the 16S rRNA gene specific for the bacterium Parachlamydia acanthamoebae, the size of 88 base pairs. Verification of the analytical specificity of the PCR test system was confirmed by amplifying 9 control DNA of members of the order Chlamydiales: Waddlia chrondophila, Clavochlamydia salmonicola, Piscichlamydia salmonis, Chlamydia avium, Chlamydia pecorum, Chlamydia abortus, Chlamydia psittaci, Chlamydia suis, Chlamydia caviae. The demand for such test systems is due not only to their use in scientific research to isolate and study individual strains, to conduct epizootic monitoring, but also for the purpose of effective treatment, since distinct species have diverse sensitivity or even insensitivity to different types of antibiotics and vary in the source of infection and mode of transmission.