Conjunctivitis due to Chlamydophila felis (Chlamydia psittaci feline pneumonitis agent) acquired from a cat: case report with molecular characterization of isolates from the patient and cat.

Mixed infections with porcine Chlamydia trachomatis/pecorum and infections with ruminant Chlamydia psittaci serovar 1 associated with abortions in swine.

Amplification primers and methods for specific amplification and detection of a rnpB gene sequence are disclosed. The primer-target binding sequences are useful for amplification and detection of organisms of the Chlamydiaceae family in a variety of amplification and detection reactions.

Amplification and detection of organisms of the chlamydiaceae family

Laboratory diagnosis of Chlamydophila psittaci, an important turkey respiratory pathogen, is difficult. To facilitate the diagnosis, a nested PCR-enzyme immunoassay (PCR-EIA) was developed to detect the Cp. psittaci outer membrane protein A (ompA) gene in pharyngeal swabs. The fluorescein-biotin labelled PCR products were immobilized on streptavidin-coated microtiter plates and detected with anti-fluorescein peroxidase conjugate and a colorimetric substrate. An internal inhibition control was included to rule out the presence of inhibitors of DNA amplification. The diagnostic value of the ompA nested PCR-EIA in comparison to cell culture and a 16S-rRNA based nested PCR was assessed in pharyngeal turkey swabs from 10 different farms experiencing respiratory disease. The sensitivity of the nested PCR-EIA was established at 0.1 infection forming units (IFU). Specificity was 100%. The ompA nested PCR-EIA was more sensitive than the 16S-rRNA based nested PCR and isolation, revealing 105 out of 200 (52.5%) positives against 13 and 74 for the latter two tests, respectively. Twenty-nine (23.8%) out of 122 ompA PCR-EIA negatives showed the presence of inhibitors of DNA amplification, although 27 of them became positive after diluting (1/10) the specimens in PCR buffer or after phenol-chloroform extraction and subsequent ethanol precipitation. The present study stresses the need for an internal control to confirm PCR true-negatives and demonstrates the high prevalence of chlamydiosis in Belgian turkeys and its potential zoonotic risk. The ompA nested PCR-EIA described here is a rapid, highly sensitive and specific diagnostic assay and will help to facilitate the diagnosis of Cp. psittaci infections in both poultry and man.

/pdf/use-of-a-nested-pcr-enzyme-immunoassay-with-an-internal-slw9s9ic7o.pdf

Use of a nested PCR-enzyme immunoassay with an internal control to detect Chlamydophila psittaci in turkeys.

Foodborne diseases, in particular those related to meat and meat products, have recently become a matter of great public concern. Sheep and goat meat can transmit infections and diseases either through handling during preparation procedures or as a result of ingestion by the consumer. The authors highlight the second route of contamination in relation to meat and meat products from small ruminants in European countries. Among the most important diseases transmitted by mutton and goat meat, toxoplasmosis remains the greatest threat, particularly in immuno-compromised people and in pregnant women. Other pathogens which may be associated with the consumption of meat from small ruminants include: Clostridium perfringens, Cryptosporidium parvum and Campylobacter jejuni. As with other ruminant species, Escherichia coli O157:H7 can be considered as an emerging pathogen, for which control efforts must be made. The classical zoonoses (brucellosis, Q fever, hydatidosis) are also presented here, although the major source of contamination for these diseases remains contact with infected animals or the handling of carcasses. The fact that the association of foodborne diseases with mutton and goat meat is less frequent than with the meat of other animal species should be noted, for the following reasons: a) lower levels of production; b) less intensive production, leading to a weaker microbial contamination; c) mutton and goat meat are subjected to processing less often than other meats; d) the usual boiling or cooking processes.

Public health hazards from small ruminant meat products in Europe

Sequence analyses 5' ends of the 60-kDa cysteine-rich outer membrane protein genes (Omp2) of Chlamydia psittaci and Chlamydia pecorum strains indicate that these species have approximately 70% nucleotide identity. On the basis of this sequence information, PCR primers were designed to allow the specific amplification of DNA extracted from C. psittaci S26/3 (abortion strain), P94/1 (pigeon strain), and C. pecorum W73 (fecal strain) in one reaction tube. By using nested reactions (with primers PCR-D1 and PCR-D2 followed by the specific primers and PCR-D2), 0.6, 0.2, and 8 inclusion-forming units of S26/3, P94/1 (both diluted in tissue culture-negative placental material), and W73 (diluted in culture-negative fecal material) per ml, respectively, were detected. The differentiation of C. psittaci and C. pecorum strains of ovine and bovine origins was carried out, and the results were in agreement with those obtained from AluI restriction enzyme analysis of DNA amplified from corresponding strains by PCR. This approach allows the simultaneous detection and typing of C. psittaci and C. pecorum strains and the identification of samples containing both species.

Differentiation of Chlamydia psittaci and C. pecorum strains by species-specific PCR.

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Differentiation of Chlamydia psittaci and C. pecorum strains by species-specific PCR.