METHOD FOR DIFFERENTIAL DIAGNOSIS OF CHLAMYDIA SPECIES, CHLAMYDOPHILA PNEUMONIAE AND PATHOGENS OF ZOONOTIC CHLAMYDIOSIS Russian patent published in 2004 - IPC

FIELD: medicine, biology, molecular biology, biochemistry.

SUBSTANCE: invention proposes a method for carrying out differential diagnosis of representatives of family Chlamydiaceae. Method involves isolation of pathogen DNA from clinic material followed by amplification of pathogen DNA using polymerase chain reaction (PCR) in the real-time regime. Method involves carrying out the multiplex PCR with a single primers pair CM1: 5'-CAG-GAC-ATC-TTG-TCT-GGC-TT-3' and CM2: 5'-CAA-GGA-TCG-CAA-GGA-TCT-CC-3' to 5'-end of gene omp1 fragment and four probes: 5'-JOE-ATCCTGCTGAACCAAGCCTTATGAT-3'-BHQ1 for Chlamydia spp., 5'-ROX-ACCCTTCTGATCCAAGCTTATTAATTGAT-3'-BHQ2 for Ch. pneumoniae, 5'-Cy5-CCAGCTGAACCAAGTTTATTAATCGATG-3'-BHQ3 for Ch. psittaci, Ch. abortus, Ch. felis and 5'-FAM-CGGAACGATGCCATTCTGCTTAT-3'-BHQ1 for internal standard of PCR. The increment of fluorescence of corresponding fluorophores in the sequential cycles of amplifications provides to prove the presence of one or some among enumerated pathogens in analyzed DNA sample. The proposed method provides carrying out simultaneous detection and differentiation of all species of Ch. trachomatis, Ch. pneumoniae that are pathogens for humans and pathogens of zoonotic infections Ch. psittaci, Ch. abortus and Ch. felis, and to reduce contamination risk with PCR products and obtaining false-positive results due to absence of step in electrophoretic analysis of PCR-products that provides reducing labor intensity and analysis time.

EFFECT: improved diagnosis method.

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