METHOD OF DIAGNOSIS OF CHLAMYDIOSIS INFECTION (VARIANTS) Russian patent published in 2003 - IPC

FIELD: biotechnology, biology, medicine, veterinary science. SUBSTANCE: invention proposes three developed method of detection of representatives of family Chlamydiaceae involving detection of chlamydium DNA by carrying out polymerase chain reaction (PCR) using oligonucleotide primers. Two methods involve carrying out one-step PCR using in the first method primers: 5'-CAAACTCATGAGA- CGAG(CT)AGT-3' (I) and 5'-CTTTCAATGTTACAGAAAACTCTA- CAG-3' (II) and in the second method primers: 5'-ATGCAAAATTAC- TGT(AT)TGGGTAAA-3' (I) and 5'-CTTCAATGTTACAGAAAACT- CTACAG-3' (II). According to the third method reaction is carried out for two step: the first involves the use of primers: 5'-CAAACTCATCA- GACGAG(CT)AGT-3' (I) and 5'-CTTTCAATGTTACAGAAAACTC- TACAG-3' (II) and the second step involves the use of primers: 5'-CTT- TCAATGTTACAGAAAACTCTACAG-3' (II) and 5'-ATGCAAAAT- TACTGT(AT)TGGGTAAA-3' (III). In each 32 cycles of polymerase chain reaction are carried out that involve one cycle of denaturation stage for 4 min at 95 C and stages of annealing for 2 min at 60 C, 30 cycles of elongation stage for 1 min at 72 C, denaturation stage for 40 s at 95 C and annealing stage for 30 s at 60 C and one cycle of elongation stage for 4 min at 72 C. The use of these methods allows to carry out specific assay the chlamydia presence in sample being independently of species, species combination and the presence of species relates to chlamydia. EFFECT: improved method of diagnosis. 6 cl, 3 ex