Detection and strain differentiation of Chlamydia psittaci mediated by a two-step polymerase chain reaction.

AUTOR(ES)

Kaltenboeck, B

RESUMO

Specific and sensitive amplification of major outer membrane protein (MOMP) gene DNA sequences of Chlamydia psittaci was achieved in a two-step polymerase chain reaction. First, oligonucleotide primers specific for 5' and 3' nontranslated regulatory regions of the MOMP gene were used in a polymerase chain reaction to amplify a DNA fragment of approximately 1,400 bp. A portion of this DNA fragment was amplified in a second reaction using a degenerate oligonucleotide primer specific for a DNA sequence contained within the 1,400-bp DNA fragment and one of the first-step primers. This method detected 10 cognate chlamydial genomes. C. psittaci MOMP genes from two avian strains and from mammalian serovars 1, 7, and 8 were amplified and analyzed by restriction endonuclease digestion. MOMP genes from mammalian serovars 2 through 6 and 9 and from strains of C. trachomatis and C. pneumoniae could not be amplified. Restriction endonuclease analysis with HaeIII indicated a close relationship between C. psittaci strains of avian and mammalian serovar 1 lineage, while those of mammalian serovars 7 and 8 exhibited distinct restriction patterns. DNA sequences corresponding to the mammalian serovar 1-wild type parakeet MOMP genotype of C. psittaci were detected in two of seven milk samples from cases of bovine mastitis.

Documentos Relacionados

• Detection of plasmid DNA from all Chlamydia trachomatis serovars with a two-step polymerase chain reaction.
• Detection of Mycobacterium tuberculosis in clinical samples by two-step polymerase chain reaction and nonisotopic hybridization methods.
• Simple and rapid two-step polymerase chain reaction for diagnosis of Pneumocystis carinii infection.
• Amplification and sequence analysis of DNA flanking integrated proviruses by a simple two-step polymerase chain reaction method.
• Detection of Chlamydia pneumoniae by polymerase chain reaction.