Isolation Of Chlamydia Research Articles

Geomapping of chlamydia and gonorrhoea in Birmingham

To investigate if the core population hypothesis is applicable to patients with genital chlamydia infections. Retrospective cross sectional study. Two genitourinary medicine (GUM) clinics in the city of Birmingham and eight adjacent clinics. All patients with chlamydia (n = 665) or gonorrhoea (n = 584) attending between 1 October 1995 and 30 September 1996 with a postcode within the Birmingham health district. Controls were 727 patients seen in the same period with no infection. Postcodes were used to calculate population prevalence rates per 100,000 aged 15-65 in the 39 wards of the city and to estimate the socioeconomic status using the Super Profile (SP). Ethnic specific rates were also calculated. Data were obtained on gonorrhoea and chlamydia isolation from all the major laboratories of the city over the same time period. GUM clinic attenders accounted for 67.6% and 82.5% of all chlamydia and gonorrhoea isolates reported by the laboratories and that were available for our epidemiological analysis. Both infections were more common in men and in black ethnic groups. However, patients with gonorrhoea only infection were more likely to be of black ethnicity than those with chlamydia only infection (p = 0.0001) and to have different SP distribution (p = 0.0001). On logistic regression age < 20 years, male sex, black ethnicity, and living in neighbourhoods with SP J ("have nots") were predictive of both infections compared with controls. Overall chlamydia and gonorrhoea prevalence rates were 129 and 98.4 per 10(5) respectively. Corresponding rates for whites was 64.7 and 37.2 and for black ethnic groups 1105 and 1183 per 10(5) of each ethnic group. Eight adjacent wards accounted for 41% of the chlamydia and 66.5% of the gonorrhoea. In a large urban setting patients attending GUM clinics with chlamydia belong to core population groups with similar, but not identical, sociodemographic characteristics to patients with gonorrhoea infection.

Isolation and continuous growth of Chlamydia pneumoniae from arterectomy specimens.

For the purpose of collecting Chlamydia pneumoniae strains of vascular origin that could be grown continuously in vitro, a cell culture system has been established. Using different types of vascular specimens obtained from 38 patients, Chlamydia pneumoniae could be isolated in three (7.9%) cases. The strains were obtained from specimens of the carotid artery, the femoral artery and an infrarenal aneurysm of the abdominal aorta of three male atherosclerosis patients. Thus, viable Chlamydia pneumoniae strains are also present in vascular regions other than those hitherto described.

Respiratory pathogens: assessing resistance patterns in Europe and the potential role of grepafloxacin as treatment of patients with infections caused by these organisms.

Although most respiratory tract infections (RTI) are caused by viruses, various bacteria, particularly Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis, are common causes of community-acquired pneumonia, acute exacerbations of chronic bronchitis, otitis media and sinusitis. Empirical antibiotic therapy of patients with RTI must take account of the increasing prevalence of resistance among the predominant pathogens. Europe-wide susceptibility surveillance studies have revealed that resistance to penicillin and macrolides is highly prevalent among isolates of S. pneumoniae from France and Spain. Uniquely, in Italy, macrolide resistance is highly prevalent while the prevalence of penicillin resistance is low. Resistance to other antibiotic classes, including chloramphenicol, doxycycline and, in particular, co-trimoxazole, is associated with penicillin resistance in pneumococci, but resistance to the fluoroquinolones is rare. beta-Lactamase production is the principal mechanism of resistance in isolates of H. influenzae and M. catarrhalis, with fluoroquinolone resistance being detected rarely in these pathogens. In 1998 a surveillance study involving 15 European countries determined the susceptibilities of many respiratory pathogens to a range of antimicrobials, including grepafloxacin. The MIC(90) of grepafloxacin for 1251 isolates of S. pneumoniae was 0.25 mg/L, the MICs for only five strains being >2 mg/L, and 99.4% of all of the isolates tested were inhibited by concentrations </=0.5 mg/L. The MIC(90)s of grepafloxacin for 587 isolates of H. influenzae and 323 of Haemophilus parainfluenzae were 0.015 and 0.06 mg/L, respectively, while that for 509 isolates of M. catarrhalis was 0.03 mg/L. The MIC(90)s for 1164 isolates of methicillin-susceptible Staphylococcus aureus and 435 isolates of Klebsiella pneumoniae were 0.12 and 0.25 mg/L, respectively. Other studies have shown grepafloxacin to be highly active against clinical isolates of Legionella pneumophila (MIC(90) 0.015 mg/L), Mycoplasma pneumoniae (MIC(90) 0.5 mg/L) and Chlamydia pneumoniae (MICs 0.06-0.12 mg/L). Current susceptibility data indicate that fluoroquinolone resistance rates among bacterial respiratory tract pathogens are low in European countries. The enhanced potency and activity of grepafloxacin against isolates of S. pneumoniae, including those exhibiting resistance to unrelated classes of antibiotics, together with its activity against other respiratory tract pathogens, suggest that this drug has considerable potential as empirical therapy of patients with a wide range of RTI.

In-vitro activity of gatifloxacin against Chlamydia trachomatis and Chlamydia pneumoniae.

We compared the activity of gatifloxacin, a new quinolone, ofloxacin and erythromycin against five isolates of Chlamydia trachomatis and 20 isolates of Chlamydia pneumoniae, including TW183 and clinical isolates from the USA and Japan. Testing was done in cycloheximide-treated HEp-2 cells. Gatifloxacin was slightly less active against C. trachomatis and slightly more active against C. pneumoniae than ofloxacin, with MICs at which 90% of the isolates had no inclusions and minimal chlamydicidal concentrations at which 90% of the isolates had no inclusions after passage of 0.25 mg/L. Gatifloxacin was less active than erythromycin for both species.

Genomic relatedness of Chlamydia isolates determined by amplified fragment length polymorphism analysis.

The genomic relatedness of 19 Chlamydia pneumoniae isolates (17 from respiratory origin and 2 from atherosclerotic origin), 21 Chlamydia trachomatis isolates (all serovars from the human biovar, an isolate from the mouse biovar, and a porcine isolate), 6 Chlamydia psittaci isolates (5 avian isolates and 1 feline isolate), and 1 Chlamydia pecorum isolate was studied by analyzing genomic amplified fragment length polymorphism (AFLP) fingerprints. The AFLP procedure was adapted from a previously developed method for characterization of clinical C. trachomatis isolates. The fingerprints of all C. pneumoniae isolates were nearly identical, clustering together at a Dice similarity of 92.6% (+/- 1.6% standard deviation). The fingerprints of the C. trachomatis isolates of human, mouse, and swine origin were clearly distinct from each other. The fingerprints of the isolates from the human biovar could be divided into at least 12 different types when the presence or absence of specific bands was taken into account. The C. psittaci fingerprints could be divided into a parakeet, a pigeon, and a feline type. The fingerprint of C. pecorum was clearly distinct from all others. Cluster analysis of selected isolates from all species revealed groups other than those based on sequence data from single genes (in particular, omp1 and rRNA genes) but was in agreement with available DNA-DNA hybridization data. In conclusion, cluster analysis of AFLP fingerprints of representatives of all species provided suggestions for a grouping of chlamydiae based on the analysis of the whole genome. Furthermore, genomic AFLP analysis showed that the genome of C. pneumoniae is highly conserved and that no differences exist between isolates of respiratory and atherosclerotic origins.

Activity of grepafloxacin and other fluoroquinones and newer macrolides against recent clinical isolates of Chlamydia pneumoniae

Chlamydia pneumoniae is a frequent cause of community-acquired respiratory tract infection including pneumonia and bronchitis. Quinolones have attracted interest as potential therapy for community-acquired respiratory tract infections because they are active against a wide range of pathogens including C. pneumoniae and Mycoplasma pneumoniae. The in vitro susceptibilities of C. pneumoniae were determined for grepafloxacin, levofloxacin, moxifloxacin, trovafloxacin, clarithromycin and azithromycin. Isolates of C. pneumoniae tested included two reference strains, TW-183 and CM-1, and 12 recent clinical isolates from adults with community-acquired pneumonia. Susceptibility testing was performed in HEp-2 cells grown in 96-well microtiter plates. The MIC was the lowest antibiotic concentration at which no inclusions were seen. The MBC was the lowest concentration which resulted in no inclusions after passage in antibiotic-free medium. Grepafloxacin was the most active quinolone tested with an MIC 50 of 0.125 mg/l, MIC 90 and MBC 90 of 0.5 mg/l. Grepafloxacin may have a role in the treatment of C. pneumoniae infections, but prospective clinical studies utilizing culture are lacking.

Infection of human endothelial cells with Chlamydia pneumoniae stimulates transendothelial migration of neutrophils and monocytes.

We have previously shown that different isolates of Chlamydia pneumoniae display heterogeneity in the in vitro stimulation of chemokines and adhesion molecules from infected human endothelial cells. In the present study, we examined the ability of different isolates of C. pneumoniae to promote transendothelial migration of neutrophils and monocytes. Human umbilical vein endothelial cells (HUVEC) were infected with low (<15)-passage C. pneumoniae isolates A-03, PS-32, and BR-393 and high (>40)-passage isolates BAL-16, TW-183, and T-2634, and levels of neutrophil and monocyte transendothelial migration were determined following 24 h of infection. Compared to mock-infected controls, significant increases in neutrophil migration were observed in response to most C. pneumoniae isolates examined (P < 0.001). Levels of monocyte migration were significantly increased in response to TW-183 and T-2634 (P < 0.001). Serial passage (>40 times) of the three low-passage isolates in HEp-2 cell cultures prior to infection of HUVEC generally resulted in the promotion of higher levels of neutrophil and monocyte transendothelial migration. These findings were compatible with differences observed in the extent of interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) stimulation between low- and high-passage A-03, PS-32, and BR-393. As opposed to C. pneumoniae, infection with C. trachomatis L2 caused only a slight increase in neutrophil transendothelial migration, which correlated with the lack of measurable IL-8 levels by this species. However, significant levels of monocyte migration were induced in response to C. trachomatis L2 despite a lack of measurable MCP-1 stimulation. C. trachomatis serovars A and E also failed to induce IL-8 and MCP-1 production in HUVEC. Results from this study indicate that the passage history of C. pneumoniae may play a role in the divergence of stimulatory activities observed among isolates in human endothelial cells. In addition, the differences observed between this organism and C. trachomatis suggest that the upregulation of IL-8 and MCP-1 in endothelial cells may be unique to C. pneumoniae.

Transmission of Chlamydia trachomatis and Neisseria gonorrhoeae among men with urethritis and their female sex partners.

Transmission of Chlamydia trachomatis and Neisseria gonorrhoeae among infected men and their female sex partners was examined using a design enhancing the likelihood that spread was directed from men to women. Chlamydia culture-negative specimens were examined using DNA amplification tests. Infection rates in women exposed to male sex partners with Chlamydia only were 65% (20/31) and with gonorrhea only were 73% (33/45). Infection of women by either agent was not influenced by the number of sexual exposures to or coinfection in men. There was a 98% (40/41) concordance of N. gonorrhoeae isolates among partners by auxotype and serovar. Chlamydia isolates were serotyped using ELISA and immunofluorescence testing and confirmed by nested polymerase chain reaction: 50% (6/12) of men and 57% (8/14) of women yielded mixed serovars. Sixty-four percent of pairs (9/14) were infected with identical serovars and an additional 28% shared at least one serovar. Multiple serovars of C. trachomatis, but not of N. gonorrhoeae, were common in sex partners and exchanged frequently.

Preclinical models for Chlamydia pneumoniae and cardiovascular disease: hypercholesterolemic mice

Seroepidemiologic studies have shown an association of Chlamydia pneumoniae antibody with atherosclerosis. Compelling additional evidence has accumulated, in that the organism has been found within atherosclerotic lesions throughout the arterial tree by multiple methods. C. pneumoniae has also been isolated from coronary and carotid atheromatous plaques. Although these studies support a potential role for C. pneumoniae in atherogenesis, confirmation of a causal relationship requires the use of animal models and intervention studies. We have focused on the evaluation of mouse models to address the hypothesis that, following upper respiratory tract infection, lung macrophages are infected, disseminate to the aorta, and alter the onset or progression of atherogenesis. ApoE-deficient knock-out and C57BL/6J mice were used. The apoE-deficient mouse develops atherosclerotic lesions spontaneously on a regular diet in a time- and age-dependent manner. This knock-out strain was developed on the background of the C57BL/6J mouse, which only develops atherosclerosis on a high-fat/high-cholesterol diet. To investigate whether infected macrophages constitute a vehicle for dissemination of C. pneumoniae in vivo, mice were inoculated intranasally or intraperitoneally. The organism was detected in harvested alveolar and peritoneal macrophages at all time points following intranasal and intraperitoneal inoculations, respectively, and in peripheral blood mononuclear cells following inoculation by both routes. In another experiment, alveolar and peritoneal macrophages from intranasally and intraperitoneally inoculated mice were adoptively transferred by intraperitoneal injection to uninfected mice. Subsequently, C. pneumoniae was detected in lung, spleen, abdominal lymph nodes and/or thymus of recipient mice. In control experiments, UV-inactivated C. pneumoniae DNA was not detected in alveolar or peritoneal macrophages beyond 5 min after inoculation in vivo. These cumulative results demonstrate that C. pneumoniae infects macrophages in vivo and that macrophages can serve as a vehicle for dissemination to other sites. To answer the question of whether the organism disseminates to and persists in the aorta, 8-week-old mice were infected intranasally. Following single or multiple inoculations in apoE-deficient mice, C. pneumoniae was detected in the lung and aorta for 20 weeks postinfection. In contrast, in C57BL/6J mice, the organism did not persist in the aorta following a single intranasal inoculation, but could be detected up to 7 weeks postinfection in multiply inoculated mice. Significantly, in apoE-deficient mice with developed atherosclerotic lesions, the organism was found in foam cells within the lesions by immunocytochemical staining. These studies show that persistent C. pneumoniae infection occurs in atherosclerotic lesions in the aorta in the apoE-deficient knock-out mouse model. Infection of the aorta also occurred in C57BL/6J mice but was more transient. Both models should be useful in studying the pathogenic role of C. pneumoniae in atherogenesis and for determining if therapy prevents dissemination of infection. To this end, we have evaluated the susceptibility of C. pneumoniae to roxithromycin, a new macrolide, in vitro in cell culture and in vivo in the apoE-deficient mouse model. In vitro results compare favorably with other new macrolides, quinolines, and tetracyline. Preliminary studies in the apoE model of persistent infection suggest that the organism is detected less frequently in the lung and aorta by PCR following treatment. These promising preliminary studies warrant further investigation.

Experimental infection of pregnant ewes with Chlamydia pecorum.

Pregnant ewes were infected in midpregnancy with three isolates of Chlamydia pecorum derived from the feces of healthy lambs from three different farms. Oral infection, alone or together with Fasciola hepatica, did not result in tissue invasion, since all placental and fecal samples were negative for chlamydiae. Intravenous infection resulted in placental infection in 16 of 18 ewes in that chlamydiae were cultured from placentas or vaginal swabs. Two ewes bore dead lambs after a shortened gestation time. The chlamydiae isolated were all C. pecorum. There were no significant differences between the weights of the lambs from the infected groups and those from uninfected control ewes. Most ewes showed no serological evidence of infection by the complement fixation test; therefore, it is unlikely that the enteric subtype of C. pecorum is responsible for the cross-reactions sometimes seen in flocks being tested for C. psittaci infection.

Reactions of polyclonal and neutralizing anti-p54 monoclonal antibodies with an isolated, species-specific 54-kilodalton protein of Chlamydia pneumoniae.

A recently described 54-kDa protein has been detected in six type strains and three patient isolates of Chlamydia pneumoniae by immunoblotting with sera from patients positive for antibodies to C. pneumoniae by the microimmunofluorescence test. This protein was not found in either C. trachomatis E or C. psittaci Z 432 as an antigen, confirming its species specificity. The 54-kDa protein was isolated by continuous-elution electrophoresis and immunoglobulin G monoclonal antibodies (MAbs) against the isolated antigen were produced. MAb 8B11E6 reacted only with the 54-kDa band of C. pneumoniae and not with C. trachomatis E or C. psittaci in a Western immunoblot assay. This antibody was purified and tested for neutralizing activity together with three additional anti-p54-active MAbs (8B11E6, 8B11B4, and 10F1C1). In Buffalo green monkey cells, all of the MAbs significantly reduced the infectivity of C. pneumoniae elementary bodies, whereas no neutralizing activity could be observed with C. trachomatis E or C. psittaci Z 432. These results not only confirm the species specificity of the 54-kDa protein but also indicate that this protein might play an important role in the pathogenesis of C. pneumoniae infection. Furthermore, the results suggest a possible protective role of anti-p54 antibodies in an adaptive immune response.

A novel method for isolation of Chlamydia pneumoniae by treatment with trypsin or EDTA.

To establish a novel method for the efficient isolation of Chlamydia pneumoniae, experiments were performed to determine the effects of EDTA or trypsin treatment of C. pneumoniae on its adsorption and inclusion body formation. Treatment of C. pneumoniae with 0.1% trypsin or 1 mM EDTA significantly increased inclusion body-forming activity from 8,000- to 10,000-fold higher than that of the control. C. pneumoniae was successfully isolated in cultured cells which were inoculated with clinical specimens after treatment with 0.1% trypsin.

Species identification of Chlamydia isolates by analyzing restriction fragment length polymorphism of the 16S-23S rRNA spacer region.

The genetic diversity of the 16S-23S rRNA spacer region of 12 Chlamydia pneumoniae isolates, 7 Chlamydia trachomatis isolates (human biovars: the trachoma serovars B and C, the urogenital serovars D, E, and F, and the lymphogranuloma venereum serovar L2; and a mouse biovar), 6 Chlamydia psittaci isolates (5 avian isolates and 1 feline isolate), and one Chlamydia pecorum isolate was studied. The 16S-23S rRNA spacer region was amplified by PCR and digested with the restriction enzymes MseI, PstI, AluI, BglII, NlaIV, and BclI. All 26 isolates could be amplified by using one genus-specific primer pair, yielding an amplicon with a size of 803 bp. The analytical sensitivity of the PCR assay was < or = 100 inclusion-forming units per reaction. Digestion with MseI or AluI made it possible to differentiate the four species from each other, the C. trachomatis human biovars from the mouse biovar, and the C. psittaci avian isolates from the feline isolate. The MseI profiles were more distinct than the AluI profiles. Phylogenetic analysis of the results for all enzymes combined supported the current classification of four Chlamydia species: C. pneumoniae, C. trachomatis, C. psittaci, and C. pecorum. Phylogenetic analysis also suggested subdivision of isolates of C. trachomatis and C. psittaci into subgroups that coincide with their host range. In conclusion, we have developed an easy and rapid method for species and subspecies identification of Chlamydia based on restriction fragment length polymorphism analysis of the 16S-23S rRNA spacer region.

Studies on the Chlamidia Psittaci Infection in Birds And Humans

ABSTRACTChlamydia psittaci infection (chlamydosis, psittacosis-ornithosis is a naturally occurring systemic disease of birds. It is recognized by thy World Health Organization as particularly dangerous for humans zooantroponosis. In the work are presented etiological, epidemiological, clinical and morphological studies on recent outbreaks of chlamydiosis in waterfowl, hens and other avian species, as well as in persons occupationally involved in the poultry industry. Employed were the methods of complement fixation test, isolation and cultivation of chlamydiae in chicken embryos, light and electron microscopy, histology, epidemiological analysis. Investigated were a total of 3707 birds and 408 persons involved in poultry industry. Chlamydial CF-antibodies were detected in ducks (16.49–25.20 %) in 13 foci, in geese (12.00–40.75 %) in 6 foci, hens (13.42–41.17 %) in 3 foci, as well as in pigeons, pheasants, parrots and magpies. Eight Chlamydia psittaci strains were isolated. Serological data for chlamydial infection were obtained in diseased and contact persons. The direct electron microscopy of C. psittaci in pathological materials from birds is a valuable method. Presented is ultrastructural characteristic of the chlamydiae, as well as the histological findings in the tissues of birds with chlamydiosis. The observed clinical course among the birds varied from acute to inapparent forms. The human clinical cases were manifested with expressed adynamics, febrile reactions, respiratory signs, interstitial pneumonia. The reported results may contribute for better understanding of the problem and for more efficient diagnosis and control of this zooantroponosis.

Isolation of Chlamydia pneumoniae from the coronary artery of a patient with coronary atherosclerosis. The Chlamydia pneumoniae/Atherosclerosis Study Group.

Atherosclerosis is pathologically similar to a chronic inflammatory response. Recent reports have suggested that Chlamydia pneumoniae may play a role in the pathogenesis of atherosclerosis. To determine, by using various detection methods, whether C. pneumoniae is present in the coronary arteries of patients with coronary atherosclerosis. Multicenter investigation. The Jewish Hospital Heart and Lung Institute in Louisville, Kentucky, and several laboratories. 12 patients seeking heart transplantation. Culture for C. pneumoniae was done in HEp-2 cell monolayers. Other methods of detection included polymerase chain reaction (PCR) assay, immunocytochemistry, transmission electron microscopy, and in situ hybridization. Chlamydia pneumoniae was cultured from atherosclerotic plaques in one patient with severe coronary artery disease. The organism was found in the atheromas of this patient by PCR assay, immunocytochemistry, electron microscopy, and in situ hybridization. In addition, at least one testing method showed C. pneumoniae in coronary artery tissue in six of nine additional patients with coronary atherosclerosis. This study provides direct evidence of the presence of viable C. pneumoniae in atheromatous lesions. A chronic inflammatory response caused by a persistent infection of the coronary arteries may explain the link between C. pneumoniae and atherosclerosis.

Effect of hydrocortisone succinate on growth of Chlamydia pneumoniae in vitro.

We examined the effect of hydrocortisone succinate on the growth of three isolates of Chlamydia pneumoniae in vitro. There was a significant increase in the number of inclusions seen in two of the C. pneumoniae strains in the presence of hydrocortisone. There was no significant increase in the number of inclusions with various concentrations of hydrocortisone over time. The addition of hydrocortisone did not affect the in vitro activities of azithromycin, erythromycin, and doxycycline against C. pneumoniae.

In vitro activities of azithromycin and doxycycline against 15 isolates of Chlamydia pneumoniae.

Fourteen isolates of Chlamydia pneumoniae, 12 from clinically ill patients and 2 from subjectively healthy individuals from an area within a 400-km proximity of Gävle, Sweden, and strain IOL-207, originally from the eye of an Iranian child, were tested for susceptibilities to the antibiotics doxycycline and azithromycin. MICs and minimum chlamydiacidal concentrations were found to correlate well with values reported earlier by other investigators. In addition to MIC and minimum chlamydiacidal concentration testing, testing for the viability of C. pneumoniae after exposure to antibiotic concentrations as high as 50 mg/liter was carried out by passaging antibiotic-treated, infected cell cultures four times in the absence of antibiotics. It was found that all Chlamydia strains were viable after four passages, regardless of antibiotic concentration in the cell culture.

Bacteriologic diagnosis of respiratory tract infection

The use of the Gram stain for determination of the presence of bacteria in respiratory secretions is described. Success of the procedure depends on the use of fresh samples of high cellular quality. The Gram stain can be used to distinguish the presence (among other organisms) of Streptococcus pneumoniae, Haemophilus influenzae, Branhamella catarrhalis enterobacteria and Pseudomonas. This paper also discusses the use of sputum samples for bacteriologic culture and the value of bacterial counts in expectorated sputum, and describes the use of transtracheal aspiration and transthoracic aspiration puncture. Finally, it considers the optimum procedures for the testing of antibiotic susceptibility of presumptive pathogens isolated from the lower respiratory tract.

A randomized trial of prophylactic doxycycline for curettage in incomplete abortion

To determine whether prophylactic doxycycline at suction curettage for incomplete abortion decreases the rate of postoperative pelvic infection. We randomized 240 patients to receive intravenous doxycycline or placebo at curettage. Cervical specimens for gonorrhea and chlamydia were obtained preoperatively. Two weeks post-procedure, we evaluated all patients for infectious morbidity and repeated gonorrhea and chlamydia cultures. Statistical analysis used Mann-Whitney U test, McNemar test, or Fisher exact test, as appropriate. There were no statistically significant differences in age, parity, gestational age, history of sexually transmitted disease, pelvic inflammatory disease, or multiple sex partners between the doxycycline and placebo groups. Preoperative gonorrhea or chlamydia isolates were positive in five (4.2%) and six (5%) of 120 doxycycline patients and four (3.3%) and eight (6.6%) of 120 controls (not significant). All preoperative gonorrhea isolates remained positive postoperatively. Seven (5.8%) controls had positive postoperative chlamydia isolates, as did one (0.8%) in the doxycycline group (P = .06). We diagnosed eight (6.6%) of 120 doxycycline patients and seven (5.8%) of 120 controls with infectious morbidity (not significant). In our population of patients with incomplete abortion, the prevalence of gonorrhea and chlamydia was low, and prophylactic doxycycline did not decrease the rate of postoperative febrile morbidity.

Development and evaluation of an indirect ELISA to detect antibodies to abortion strains of Chlamydia psittaci in sheep sera

A novel indirect enzyme-linked immunosorbant assay (ELISA) for antibodies against abortion strains of Chlamydia psittaci (C. psittaci) has been developed. The antigen used was chlamydial elementary bodies treated sequentially with N-lauroyl sarcosine and n-octyl-β-D-glucopyranoside and finally solubilized with N-lauroyl sarcosine and dithiothreitol. Treating the antigen with sodium periodate after coating of the plates increased the specificity for antibodies to abortion strains. The test was evaluated initially with sera from experimentally infected sheep and an uninfected control group. These sheep were monitored for lambing performance and infection status. When used in conjunction with the indirect micro-immunofluorescence test (MIF), the ELISA was able to identify as negative all twenty-five sera from ewes that had no typical placental lesions and identified as positive twenty of twenty-one sera from infected ewes that had either typical placental lesions or had been found positive by isolation of chlamydia in cell culture. The combination of ELISA and MIF was also able to discriminate correctly groups of sera from six flocks with a history of infection from four known uninfected flocks.