SPECIFICITY DETERMINATION AND VALIDATION OF THE PCR TEST-SYSTEMS DEVELOPED FOR DIAGNOSING CHLAMYDIOSIS OF DOMESTIC CARNIVORES
T. M.Tsivenko, I. M.Ksyonz
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The NAAS Institute for Pig Breeding and Agro-Industrial Production
Shvedska Mogyla Str., 1, Poltava, 36013, Ukraine
The results are presented on determining the analytic specificity and validation of the self-design PCR test-systems for indicating bacteria of the Chlamydia genus in biological samples taken from cats and dogs. The developed PCR test-systems include pairs of oligonucleotide primers flanking the areas of MOMP genes DNA of Chlamydia felis and RNase P RNA of Chlamydia abortus, Chlamydia pecorum and Chlamydia psittaci. The purpose of the study was to determine the analytical specificity of the designed PCR test-systems for diagnosing Chlamydial infections of domestic carnivores and their validation.Validation of the self-design test-systems was performed by the method of comparing them with other PCR test-systems for indication and species differentiation of the Chlamydia genus bacteria.
The total of 19 biological material samples served as the material to be tested (control DNA of the four Chlamydia species: C. felis, C. abortus, C. pecorum, C. psittac,i and Chlamydia isolates extracted from epithelial scrapes of the conjunctival mucosa, groin, prepuce, 20%-suspensions of visceral biopsy materials from domestic dogs and cats, and faeces and epithelial scrapes from the rectum mucosa of wild carnivores belonging to Felidae and Canidae families).
Analysis of the amplification products electrophoregrams of the mentioned biological samples obtained by using two designed test-systems and three other PCR test-systems for indicating Chlamydia genus bacteria, as well as the multiplex PCR test-system for differentiating Chlamydia as to their species, proves their absolute adequacy.
The results of the study prove specificity of the mentioned diagnosticums, and respectively, the possibility of their using for diagnosing Chlamydial infections of domestic carnivores.
Keywords: CHLAMYDIOSES, DOMESTIC CARNIVORES, PCR TEST-SYSTEM, OLIGONUCLEOTIDE PRIMERS, MOMP, RNASE P RNA
