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PubMed1992

Enzyme immunoassay for anti-treponemal IgG: screening or confirmatory test?

AIMS: To review the performance of the Venereal Diseases Research Laboratory (VDRL) test and the Treponema pallidum haemagglutination assay (TPHA) as a combined screen for syphilis to provide a baseline for assessing screening by anti-treponemal IGG EIA. METHODS: Between 1980 and 1987 all serum samples were screened by both VDRL and TPHA tests. The FTA-ABS test was also used in suspected early primary syphilis, or when one of the other tests was positive. A positive result in a screening test was confirmed by quantitative testing. From 1988 all specimens were screened with an enzyme immunoassay (Captia Syph G) as a single screening test. RESULTS: Of the 44 primary, 47 secondary, and 38 early latent cases of syphilis, the VDRL and TPHA detected 32 (73%) and 31 (71%) of the primary cases; the combination detected 37 (84%). All 85 cases of cases of secondary and early latent infection were reactive in the TPHA test, whereas the VDRL was reactive in only 68 (80%). EIA had a reported sensitivity of 82% for primary infection. CONCLUSIONS: EIA can be used as a single screening test for detecting early syphilis because its results are comparable with those of the combined VDRL and TPHA tests. The conventional VDRL test should not be used as a single screening test.

Antibodies, Bacterial↗
PubMed2005

Oral bacteria in the occluded arteries of patients with Buerger disease.

OBJECTIVE: Recent studies have suggested that infectious organisms play a role in vascular diseases. In this study, to explore a possible link between oral infection and Buerger disease, we investigated whether oral (periodontal) bacteria were present in occluded arteries removed from patients with characteristic Buerger disease. METHODS: Fourteen male patients with a smoking history who had developed characteristics of Buerger disease before the age of 50 years were included in this study. Occluded arteries, including superficial femoral (n = 4), popliteal (n = 2), anterior tibial (n = 4), and posterior tibial (n = 4) arteries, were removed and studied. A periodontist performed a periodontal examination on each patient and collected dental plaque and saliva samples from them at the same time. The polymerase chain reaction method was applied to detect whether seven species of periodontal bacteria--Porphyromonas gingivalis, Tannerella forsythensis, Treponema denticola, Campylobacter rectus, Actinobacillus actinomycetemcomitans, Prevotella intermedia , and Prevotella nigrescens--were present in the occluded arteries and oral samples. In addition, arterial specimens from seven control patients were examined by polymerase chain reaction analysis. RESULTS: DNA of oral bacteria was detected in 13 of 14 arterial samples and all oral samples of patients with Buerger disease. Treponema denticola was found in 12 arterial and all oral samples. Campylobacter rectus, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, and Prevotella nigrescens were found in 14% to 43% of the arterial samples and 71% to 100% of the oral samples. A pathologic examination revealed that arterial specimens showed the characteristics of an intermediate-chronic-stage or chronic-stage lesion of Buerger disease. All 14 patients with Buerger disease had moderate to severe periodontitis. None of the control arterial samples was positive for periodontal bacteria. CONCLUSIONS: This is the first study to identify oral microorganisms in the lesions of Buerger disease. Our findings suggest a possible etiologic link between Buerger disease and chronic infections such as oral bacterial infections.

Adult↗
PubMed1976

Experimental syphilis vaccines in rabbits. I. Differential protection with an adjuvant spectrum.

This study shows that 1 per cent. glutaraldehyde fixation of unwashed Treponema pallidum, Nichols pathogen, preserves the integrity of the outer envelope as well as the ruthenium-red (RR) staining extracellular slime layer. Since other researchers wash their treponemal suspensions before preparing their vaccines, it is likely that the vaccine preparations used in this study contain antigens not tested elsewhere. In an attempt to eliminate the requirement of large numbers of injections to achieve any protection, the rabbits in this study were injected with glutaraldehyde-fixed T. pallidum (GFTP) only once, with or without one of a spectrum of adjuvants, and 5 months later were challenged intradermally or intratesticularly with freshly harvested pathogenic treponemes. Data of time of lesion appearance and severity of lesions reveal a differential protective effect of the various vaccine preparations when compared to the response of non-vaccinated controls. Vaccines which included phytohaemagglutinin (PHA) or alumina C gel (ALC) as adjuvants provided some degree of protection, whereas vaccines which contained the adjuvants incomplete Freund's, zymosan, complete Freund's, polyadenylic-polyuridylic acid, or no adjuvant at all produced enhanced infections. The differential protection data indicate that various portions of the immune mechanism are either suppressed or are not stimulated by T. pallidum antigens alone, but can be made to respond with a minimum of vaccine injections.

Adjuvants, Immunologic↗
PubMed2004

Epidemiology and etiology of sexually transmitted infection among hotel-based sex workers in Dhaka, Bangladesh.

The prevalence of sexually transmitted infections (STIs) and reproductive tract infections (RTIs) among hotel-based sex workers (HBSWs) in Dhaka, Bangladesh, was studied. A total of 400 HBSWs were enrolled in the study during April to July 2002. Endocervical swabs, high vaginal swabs, and blood samples from 400 HBSWs were examined for Neisseria gonorrhoeae (by culture), Chlamydia trachomatis (by PCR), Trichomonas vaginalis (by microscopy), antibody to Treponema pallidum (by both rapid plasma reagin and Treponema pallidum hemagglutination tests), and antibody to herpes simplex virus type 2 (HSV-2) (by enzyme-linked immunosorbent assay). Sociodemographic information as well as gynecological and obstetric information was collected. Among the HBSWs, 228 women (57%) were symptomatic and 172 (43%) were asymptomatic, 35.8% were positive for N. gonorrhoeae, 43.5% were positive for C. trachomatis, and 4.3% were positive for T. vaginalis. A total of 8.5% had syphilis, 34.5% were positive for HSV-2, and 86.8% were positive for at least one RTI or STI. There was no significant difference between the prevalences of STIs among the symptomatic and asymptomatic HBSWs. These data suggested a high prevalence of STIs, particularly gonorrhea and chlamydia, among HBSWs in Dhaka.

Abortion, Spontaneous↗
PubMed1982

Serotype-specific opsonization of Treponema hyodysenteriae.

Treponema hyodysenteriae was shown to attach to mouse peritoneal cells in the absence of serum opsonins in vitro. If serotype-specific antiserum from pigs was added to the media and treponemes of that corresponding serotype were employed in the assay, the amount of attachment increased an average of 3.7 times that of the control without pig sera. However, the amount of attachment was increased an average of only 1.5 times that of the control if organisms of any noncorresponding serotype of T. hyodysenteriae were used in the assay. Since the lipopolysaccharide (LPS) extracted from T. hyodysenteriae is the basis for serotyping the treponeme, the ability of these distinct LPS types to block attachment by blocking opsonization of the organisms was tested. Attachment, using corresponding antisera and treponemes, was blocked by LPS extracted from treponemes of that serotype, but not by LPS extracted from treponemes of other serotypes. These results indicate that antibody response to T. hyodysenteriae infection in pigs is serotype-specific. Furthermore, since opsonization and subsequent attachment of bacteria to phagocytes is known to be a protective mechanism, we suggest that the LPS may be an important antigen in the stimulation of host defense against the treponeme.

Animals↗
PubMed1995

Pathology of the umbilical cord in congenital syphilis: analysis of 25 specimens using histochemistry and immunofluorescent antibody to Treponema pallidum.

Identification of Treponema pallidum in the placenta is important for diagnosis of congenital syphilis; however, spirochetes are difficult to observe in chorionic villi. To determine the sensitivity of umbilical cord examination for T pallidum, and the association of spirochetes with cord pathology, placentas were prospectively obtained from 25 women with untreated syphilis. The most common finding using hematoxylin-eosin staining was a normal-appearing umbilical cord (48%); necrotizing funisitis was the most frequent pathological lesion (36%). Spirochetes were detected using silver and immunofluorescent staining in 89% of cords, including 92% of histologically normal and 84% of abnormal cords. Three specimens showed subamnionic aggregates of spirochetes, consistent with amniotic fluid infection. Necrotizing funisitis was strongly associated with umbilical artery infection by spirochetes (P = .008). There was a 100% correlation between results of silver and immunofluorescent staining. The umbilical cord is a sensitive site for morphological confirmation of T pallidum; it is significant for the pathologist that spirochetes may often be detected in the absence of overt tissue inflammation or necrosis.

Antibodies, Bacterial↗
PubMed1990

Efficacy of azithromycin for therapy of active syphilis in the rabbit model.

Azithromycin was shown to be as effective as standard benzathine penicillin and erythromycin in the therapy of active syphilis in the rabbit model. Following production of primary chancres by intradermal inoculation of 10(6) Treponema pallidum, groups of six rabbits were treated with benzathine penicillin (200,000 units im weekly for two weeks), erythromycin base (30 mg/kg/day orally four times daily for 15 days) or azithromycin (30 mg/kg/day given orally once or twice daily for 15 days); one group was untreated. Daily darkfield (DF) microscopic examinations of chancre aspirates were conducted to identify motile organisms. Although all treated animals became DF negative prior to completion of therapy, the median time to DF negativity was longer in animals given azithromycin once daily, compared with animals receiving benzathine penicillin (P less than 0.01); no difference was seen in comparison with animals receiving erythromycin. Untreated animals remained DF positive for greater than 15 days. The mean maximum lesion diameters for all treated animals were similar and were significantly smaller than in untreated rabbits; fewer lesions ulcerated in treated than in untreated animals. Subsequent dose-ranging studies indicated that administration of lower doses of azithromycin (15 mg/kg/day given orally either once or twice daily, or 7.5 mg/kg/day given once daily) was as effective as benzathine penicillin for therapy of active syphilis in this model, though the median time to darkfield negativity was significantly longer in the azithromycin-treated animals (P less than 0.01). Persistent infection was demonstrable in lymph nodes of untreated animals, but no evidence of virulent T. pallidum was found three months following transfer of tissue from any animal treated with penicillin, erythromycin, or azithromycin.

Animals↗
PubMed1984

Evaluation of an enzyme immunoassay for IgM antibodies to Treponema pallidum in syphilis in man.

An enzyme immunoassay (EIA) for the detection of immunoglobulin M (IgM) antibodies to Treponema pallidum was investigated for specificity and sensitivity. Using the results in serum from 1192 patients with successfully treated syphilis, the assay was calculated to be about 97% specific. As in any other IgM enzyme linked immunosorbent assay (ELISA), rheumatoid factor played an important part in causing false positive results. Pre-absorption of serum with aggregated IgG was therefore necessary to perform the test. Evaluation of the results in serum from 385 patients with untreated primary, secondary, and latent syphilis as well as patients with untreated reinfections showed that the sensitivity of the assay depended on the stage of infection and varied between 98% and 93%. IgM antibody titres were about ten times higher in the EIA than in the indirect immunofluorescence assay using the IgM fractions of serum. From the results it may be concluded that the EIA is an appropriate technique not only for rapid and sensitive measurement of IgM antibodies in most patients with untreated syphilis but also for selecting treponemal IgM non-reactive patients.

Antibody Specificity↗
PubMed2003

[Prevalence and factors associated with syphilis and genital herpes in 2 groups of the female population].

OBJECTIVE: To assess the frequency of HSV-2 and Treponema pallidum markers and their relationship with characteristics of women during gynecological outpatient visits. MATERIAL AND METHODS: In 1994 and 1995, two populations of 388 and 448 women were studied in two hospitals; one in Cuernavaca, Morelos, México, and the other in Mexico City. Participants provided a blood specimen for detection of antibodies against HSV-2 and T pallidum using the Western blot technique and the VDRL and FTA-ABS tests. Also, participants answered a questionnaire on their sociodemographic characteristics and sexual behavior. The data were analyzed with the SPSS and EGRET statistical packages. RESULTS: The frequencies of HSV-2 antibodies were 28.3% in women from the first hospital, and 18.1% for those in the second. The frequencies of T pallidum antibodies were 2.3% and 1.1%, respectively. Age, marital status, education level, and number of sexual partners were associated with HSV-2 infection. CONCLUSIONS: Low frequencies were found for infection by the microorganisms studied in both groups. HSV-2 infection was associated to exposure periods, sexual behavior, and socioeconomic level. The English version of this paper is available at:http://www.insp.mx/salud/index.html.

Adult↗
PubMed1995

[Electron microscopic detection of spirochetes in dermatitis digitalis of cattle].

In typical Dermatitis digitalis (D.d.) lesions, spirochaete-like bacteria with variations in spiralization were revealed by electron microscopy. While, in the early stages of the disease, these are found to be associated with fibrillar material of keratocytes, they occur massively in vacuoles at more advanced stages. The spirochaetes carry one pair of endoflagella, originating with a hook from the poles of the bacteria. These flagellae are composed of coiled flagellating fibrils in the pole region, merging towards the centre of the bacterium. A coat of fibrils was found in association with the cytoplasmatic membrane. The winding of this coat follows and may influence the coiling of the protoplast, and is probably involved in the rapid motility of this spirochaetes, together with the flagella. Immuno-electronmicroscopy revealed an antigenic relationship with Borrelia burgdorferi, at least with regard to the regions of flagella and undulating membrane. The paper discusses: 1. The possible classification of these spirochaetes with the genus Treponema; 2. The layer of peptidoglycan occurring on the outer membrane; and 3. The keratolytic activity of spirochaetes in D.d.

Animals↗
PubMed1995

Growth of Serpulina (Treponema) hyodysenteriae under iron-restricted conditions.

Reference strains of Serpulina hyodysenteriae expressed at least three iron-regulated proteins with apparent molecular masses of > 200, 134, and 109 kDa when grown under iron-restricted conditions. Cells of S. hyodysenteriae grown under these conditions also showed increased outer membrane bleb formation when examined by electron microscopy after negative staining. S. hyodysenteriae did not use the 2 most common types of siderophore, namely catechol and hydroxamate. Western blotting with serum from a pig experimentally infected with S. hyodysenteriae B204 indicated that the 109-kDa major iron-regulated protein was expressed in vivo and was conserved among all strains tested.

Animals↗
PubMed1997

Antibody to Haemophilus ducreyi among trucking company workers in Kenya.

BACKGROUND AND OBJECTIVES: To determine the prevalence, correlates, and incidence of Haemophilus ducreyi antibodies, a cohort of East African trucking company employees was evaluated. STUDY DESIGN: Human immunodeficiency virus (HIV)-1-seronegative men working in six trucking companies in Mombasa, Kenya, were evaluated with a questionnaire and serologic testing for antibodies to H. ducreyi and other sexually transmitted pathogens. Men who were initially H. ducreyi seronegative were retested at 1 year of follow-up. RESULTS: The H. ducreyi seroprevalence among 501 men at enrollment was 26.5%. Seropositivity was significantly associated with older age, married status, years of active sex life, number of sex partners in the past year, history of unprotected sex with a prostitute in the past year, and history of alcohol intake (all P values < 0.01). Occupational travel for more than 14 days per month was also significantly associated with H. ducreyi seropositivity (odds ratio [OR] 2.1, 95% confidence interval [CI] 1.3-3.2). Using multivariate analysis, H. ducreyi seropositivity was independently associated with age, married status, history of sex with a prostitute, and history of alcohol intake. Presence of H. ducreyi antibodies was significantly associated with seropositivity to the other major genital ulcerative pathogens, Treponema pallidum (OR 4.3, 95% CI 2.2-8.3), herpes simplex virus type 2 (OR 4.9, 95% CI 2.0-11.5), and Chlamydia trachomatis (OR 3.2, 95% CI 1.5-6.9). These associations remained significant after adjusting for demographic and exposure variables. The incidence of seroconversion to H. ducreyi antibodies was 3.6 per 100 person years. CONCLUSIONS: Serologic evidence of H. ducreyi infection was common among male trucking company employees. H. ducreyi seropositivity is an objective marker of high-risk behavior and is associated with serologic evidence of other ulcerative sexually transmitted diseases.

Adolescent↗
PubMed2006

A case-control study on the association between female genital mutilation and sexually transmitted infections in Sudan.

OBJECTIVE: To assess whether the extent of female genital mutilation (FGM) influences the risk of acquiring sexually transmitted infections (STIs). DESIGN: Hospital-based case-control study. SETTING: Two obstetric/gynaecological outpatient clinics in Khartoum, Sudan, 2003-2004. POPULATION: A total of 222 women aged 17-35 years coming to antenatal and gynaecological clinics. METHODS: Women recruited for the study were divided into cases with seropositivity for Neisseria gonorrhoeae (gonococcal antibody test), Chlamydia trachomatis (enzyme immunoassay) or Treponema pallidum (Treponema pallidum haemagglutination assay) (n= 26) and controls without antibodies to these species (n= 196). Socio-demographic data were obtained and physical examination including genital examination was performed in order to classify the form of FGM. Cases and controls were compared using logistic regression to adjust for covariates. MAIN OUTCOME MEASURES: Extent of FGM and seropositivity for C. trachomatis, N. gonorrhoeae or T. pallidum. RESULTS: Of the cases, 85% had undergone the most severe form of FGM involving labia majora compared with 78% of controls (n.s.). Thus, there was no association between serological evidence of STIs and extent of FGM. The only factor that differed significantly between the groups was the education level, cases with STIs having significantly shorter education (P= 0.03) than controls. CONCLUSIONS: There is a little difference between cases and controls in regard to FGM. Having in mind the relatively small sample size, the results still indicate that FGM seems neither to be a risk factor for nor protective against acquiring STIs. This is important as argument against traditional beliefs that FGM protects against pre/extramarital sex.

Adolescent↗
PubMed1989

Evaluation of multivalent Leptospira fluorescent antibody conjugates for general diagnostic use.

Four lots of conjugate were evaluated for optimal dilution and degree of fluorescence produced with reference cultures and bovine and porcine leptospira isolates. One lot that uniformly produced better fluorescence was evaluated for sensitivity and specificity with reference cultures, isolates, culture-positive tissues, and 13 other bacterial species. Further evaluation of the conjugates was done with bovine, porcine, and ovine specimens submitted to a diagnostic laboratory. Leptospires were detected with the fluorescent antibody test (FAT) in 9 of 21 culture-positive bovine kidneys and were detected in diluted cultures when present at concentrations of 10(2)-10(3) organisms/ml. With the exception of Treponema hyodysenteriae, FAT's of other bacterial cultures produced minimal fluorescence or were negative. Positives were characterized by moderate to brilliant fluorescence of typical cell forms, and most nonspecific fluorescence was eliminated with a flazo-orange counterstain. The results indicated that the FAT utilizing multivalent conjugates could be used successfully as an additional method for diagnosis of leptospira infections.

Animals↗
PubMedDate not supplied

Intestinal spirochetosis: first cases reported in Brazil and the use of immunohistochemistry as an aid in histopathological diagnosis.

Colonization of the colon and rectum by intestinal spirochetes is detected for the first time in Brazil in 4 of 282 (1.41%) patients who had undergone sigmoidoscopy and/or colonoscopy with a histopathological diagnosis of chronic non specific-colitis. This frequency is probably underestimated, since surgically obtained specimens were not considered in the present study. Histopathological diagnosis was performed using routine stains like hematoxylin-eosin which showed the typical, of 3-microns thick hematoxyphilic fringe on the brush border of the surface epithelium, and by silver stains like the Warthin-Starry stain. Immunohistochemical procedures using two, polyclonal, primary antibodies, one against Treponema pallidum and the other against Leptospira interrogans serovar copenhageni serogroup Icterohaemorrhagiae cross-reacted with spirochetal antigen/s producing a marked contrast of the fringe over the colonic epithelium, preserving the spiral-shaped morphology of the parasite. In one case with marked diarrhea, immunohistochemistry detected spirochetal antigen/s within a cell in an intestinal crypt, thus demonstrating that the infection can be more widely disseminated than suspected using routine stains. Immunohistochemical procedures, thus, greatly facilitate the histological diagnosis of intestinal spirochetosis and may contribute to a better understanding of the pathogenesis of the disease. Transmission and scanning electron microscopy performed in one case showed that the spirochete closely resembled the species designated as Brachyspira aalborgi.

Adolescent↗
PubMed2005

Gene organization and transcriptional analysis of the tprJ, tprI, tprG, and tprF loci in Treponema pallidum strains Nichols and Sea 81-4.

The tpr gene family of Treponema pallidum subsp. pallidum, the causative agent of syphilis, has recently become the focus of intensive investigation. TprF and TprI sequences are highly conserved among different isolates and are the targets of strong humoral and cellular immune responses of the host, and immunization with a recombinant peptide from the amino terminus of these antigens has been shown to alter significantly lesion development following homologous challenge. This indicates that these antigens are expressed during infection and strongly suggests a key functionality. tprF and tprI are located immediately downstream of the tprG and tprJ genes, respectively, separated by very short intergenic spacers (55 nucleotides for G-F and 56 nucleotides for J-I). Preliminary analysis using gene-specific primers failed to amplify tprJ in the Sea 81-4 isolate. In this study, sequence and transcriptional analysis of these loci showed a similar gene organization in the Nichols and Sea 81-4 strains, a complex pattern of transcription, and the presence of G homopolymeric repeats of variable lengths upstream of the tprF, tprI, tprG, and tprJ transcriptional start sites. However, distinctive features were also identified in the Sea 81-4 isolate, including a tprG-like open reading frame in the tprJ locus, a frameshift and a premature termination in the tprG coding sequence, a longer tprG-tprF intergenic spacer, and absence of cotranscription of the tprG-tprF genes.

Amino Acid Sequence↗
PubMed2003

Systematic cloning of Treponema pallidum open reading frames for protein expression and antigen discovery.

A topoisomerase-based method was used to clone PCR products encoding 991 of the 1041 open reading frames identified in the genome sequence of the bacterium that causes syphilis, Treponema pallidum subsp. pallidum. Cloning the open reading frames into the univector plasmid system permitted the rapid conversion of the original clone set to other functional vectors containing a variety of promoters or tag sequences. A computational prediction of signal sequences identified 248 T. pallidum proteins that are potentially secreted from the cell. These clones were systematically converted into vectors designed to express the encoded proteins as glutathione-S-transferase fusion proteins. To test the potential of the clone set for novel antigen discovery, 85 of these fusion proteins were expressed from Escherichia coli, partially purified, and tested for antigenicity by using sera from rabbits infected with T. pallidum. Twelve of the 85 proteins bound significant levels of antibody. Of these 12 proteins, seven had previously been identified as T. pallidum antigens, and the remaining five represent novel antigens. These results demonstrate the potential of the T. pallidum clone set for antigen discovery and, more generally, for advancing the biology of this enigmatic spirochete.

Antigens, Bacterial↗
PubMed1981

Redox potential and survival of virulent Treponema pallidum under microaerophilic conditions.

A strongly reduced culture medium, capable of maintaining the virulence of Treponema pallidum (Nichols) for several days, was exposed to an atmosphere of 3% oxygen in nitrogen for 2-3 days before inoculation with T pallidum. By using various volumes of medium in uniform tubes a range of redox potentials (Ecal) from -94 mV to -325 mV was produced depending on the surface area-to-volume ratios of the medium. The anaerobic medium had an Ecal value of -387 mV. The medium was inoculated with T pallidum and incubated in an atmosphere of 3% oxygen. The survival of treponemes at different redox potentials was monitored by observing the retention of motility and by measuring the latent period of infection after inoculation of the cultures into the shaved backs of rabbits. Under these conditions T pallidum survived longest at low (electronegative) redox potential. An inverse linear relationship was observed between the redox potential of the culture medium and the survival of T pallidum, as measured by the time required for a 90% reduction of virulent organisms. No optimum redox potential was detected, the most electronegative medium (-325 mV, Ecal) giving the best survival.

Bacteriological Techniques↗