Importance of species-specific antigens in the serodiagnosis of Chlamydia trachomatis reactive arthritis

OBJECTIVES: To determine the most sensitive and specific method of anti-Chlamydia antibody measurement for the serodiagnosis of Chlamydia trachomatis reactive arthritis. METHODS: Immunoblotting, enzyme-linked immunosorbent assays using six synthetic peptides or recombinant antigens and a microimmunofluorescence test were used to determine the presence of IgG, IgM and IgA in serum samples from 17 patients with C. trachomatis reactive arthritis. Twenty patients with other inflammatory arthropathies without evidence of urogenital C. trachomatis infection were used as controls. RESULTS: The best association of sensitivity (76%) and specificity (85%) was obtained when IgG and/or IgA reactivity to two species-specific antigens was determined. These antigens were synthetic peptides, derived from species-specific epitopes in the variable domain IV of the major outer membrane protein (MOMP) (Labsystems, Finland) and recombinant polypeptide encoded by open reading frame 3 of the plasmid (pgp3). CONCLUSIONS: IgG and/or IgA anti-MOMP-derived peptides and anti-pgp3 could be useful for the diagnosis of probable C. trachomatis reactive arthritis.     

Prevalence of Chlamydia trachomatis in urine of male patients with ankylosing spondylitis is not increased

OBJECTIVE: To compare the prevalence of Chlamydia trachomatis infections in ankylosing spondylitis (AS) patients with controls, using DNA amplification assays in urine specimens. METHODS: The prevalence of C trachomatis infections was assessed in 32 male AS patients and 120 age and sex matched controls. Urine specimens were tested by ligase chain reaction and polymerase chain reaction. In addition, blood samples of AS patients were tested on serum antibodies to C trachomatis (IgA and IgG) by a specific peptide based solid phase enzyme immunoassay. A questionnaire was used to assess the differences in sexual behaviour and ethnic origin between the two groups. AS patients were also asked about disease characteristics. RESULTS: No significant differences were found between cases and controls in the prevalence of C trachomatis infections. No associations were found between C trachomatis antibodies and disease characteristics, except for acute anterior uveitis (AAU). Four of eight (50%) AS men positive for IgG had a history of AAU in comparison with three of 24 (12.5%) IgG negative men (OR = 7.0; 95% confidence intervals: 1.1, 44.1). CONCLUSION: The prevalence of Chlamydia trachomatis infections, as detected by commercially available DNA amplification assays in urine specimens, in AS patients is not higher compared with male controls of the same age. However, there seems to be an association between specific antibodies to C trachomatis and AAU.     

Studies on Chlamydia pneumoniae, strain TWAR infection. 2. Seroepidemiology of TWAR on healthy controls and patients with acute respiratory infections

To determine of Chlamydia pneumoniae, TWAR infection is common in Japan. The author performed a spot antibody prevalence study of adults and children living in Okayama prefecture. This study was carried out by microplate immunofluorescence antibody technique (MFA) using in situ inclusions of Chlamydia pneumoniae (TW-183), Chlamydia trachomatis (L2) and Chlamydia psittaci (Cal 10) as antigens respectively. And each chlamydial antiserum IgG, IgA, IgM titers were determined by the dilution and point for specific staining of the inclusions. The author studied people with and without evidence of acute respiratory infections, as determined by physical examination and medical history. 2050 sera of 1477 cases were collected from both groups between 1987 and 1989. The criteria for judgement the positivity of antibodies to chlamydia species are as follows: By comparing the IgG antibody titers determined with different chlamydia antigen, a case was regarded as having an antibody specific to a particular chlamydia species when the titer was highest for that strain and the titer was greater than x 64. In cases which blood was collected more than once, the point in time at which IgG showed the highest titer was used. TWAR specific antibody was detected to be 52/143 cases (36.4%) of healthy children (age 0-15) and 319/531 cases (60.1%) of healthy adults (age 16-85). It was also detected to be 134/478 cases (27.9%) of child patients with acute respiratory infections (age 0-15), and 231/325 cases (71.1%) of adult patients (age 16-95).(ABSTRACT TRUNCATED AT 250 WORDS)     

Ophthalmia neonatorum in Nairobi, Kenya: the roles of Neisseria gonorrhoeae and Chlamydia trachomatis

Among 149 consecutive infants with ophthalmia neonatorum in Nairobi, Neisseria gonorrhoeae was recovered from 43%, Chlamydia trachomatis from 13%, and both microorganisms from 4%. Three of five isolates of C. trachomatis belonged to trachoma serovars. The sensitivity and specificity of a gram-stained smear for the diagnosis of gonococcal conjunctivitis were 86% and 90%, respectively. Patients with gonococcal conjunctivitis had more purulent discharge, a higher clinical severity score, and a younger age at onset of disease. Corneal epithelial edema with superficial keratitis was present in four (16%) of 25 patients with gonococcal conjunctivitis but in none of 22 other patients (P = .07). N. gonorrhoeae or C. trachomatis was isolated from the pharynx in 11 (15%) and six (23%) cases, respectively. Oropharyngeal gonococcal infection was associated with coughing (P = .007).     

Detection of anti-Chlamydia trachomatis antibody by means of enzyme immunoassay using synthetic peptide antigen]

Newly developed diagnostic kits for the detection of Anti-Chlamydia trachomatis, Peptide-Chlamvdia (LOY: Meiji Milk Products Co., Ltd., Tokyo; for IgG and IgA), were evaluated using the microimmunofluorescence assay (MIF) as the gold standard. These results were also compared to results of testing by Sero-IPALISA and immunoblot (I-B). Detection by LOY in based on enzyme immunoassay with synthetic peptides as the antigen. Thirty serum samples from pediatric patients and 130 serum samples from gynecology patients were used. All 26 pediatric samples that were positive for Chlamydia pneumoniae IgG antibody tested negative with LOY, indicating that the presence of the antibody against C. pneumoniae did not affect the assay by LOY. For 90 gynecological samples, the total, the positive and the negative agreement rates for IgG were quite high; i.e. 87.8%, 90.0% and 70.0% (LOY vs MIF), 85.6%, 85.0% and 90.0% (Sero-IPALISA vs MIF), and 92.0%, 94.9% and 70.0% (I-B vs MIF), respectively. On the other hand, many cases of MIF (-) and LOY (+) discrepancy were seen in IgA detection. In order to better understand the basis for such disagreement. 34 serum samples were collected from patients whose cervical samples were negative for the Chlamydia group antigen based on the assay with IDEIA-Chlamydia. They were then assayed by MIF and LOY. The total, the positive and the negative agreement rates for IgG were 91.2%, 100% and 90.9%, while the total and the negative agreement rates for IgA were 88.2% and 88.2% (there were no IgA positive cases). Furthermore, 6 serum samples (1 case of MIF (+) LOY (+) and 5 cases of MIF (-) LOY (+)) were provided to determine whether LOY detects C. trachomatis specific IgA antibody. Increasing amounts of C. trachomatis serovar L2 were added to the serum samples resulting in a progressive decrease in their reactivity in the LOY assay. These results lead us to speculate that LOY can reveal even low levels of C. trachomatis specific IgA antibody. In conclusion, LOY can be used as an useful kit for detecting C. trachomatis antibody.     

A study of IgA.IgG titers for C. trachomatis in serum and prostatic secretion of chronic prostatitis

In order to study the correlation between chronic prostatitis and C. trachomatis, IgA and IgG titers for C. trachomatis in serum and prostatic secretion were measured using the indirect immunoperoxidase assay (Ipazyme Chlamydia). 1) Positive rate of IgA and IgG titers in serum and prostatic secretion was higher in cases of chronic prostatitis than that in cases of suspected chronic prostatitis or the normal male group. 2) In cases of chronic prostatitis, IgA titers showed a tendency to be high in prostatic secretion, and IgG titers showed the same tendency in serum. This result was considered C. trachomatis infection was appeared local immunization in prostate. 3) After treatments with DOXY for cases of positive IgA titers in prostatic secretion, IgA titers of those cases were decreased and inflammations of prostate were cured. 4) The positive rate of IgA and IgG titers in serum was higher in wives with IgA positive cases than in those with the IgA negative. Those results suggested that chronic prostatitis was correlated by C. trachomatis.     

The infection rate of Chlamydia trachomatis in young adult men without urogenital symptoms--screening test using urine sediments

The detection of C. trachomatis antigen in first-voided urine sediments has recently been achieved by means of IDEIA CHLAMYDIA (IDEIA, Novo Nordisk), an EIA kit using monoclonal antibodies. Therefore, this kit was used as a screening test to examine the infection rate of C. trachomatis in young adult men without symptoms. The titers of serum IgA and IgG antibodies to C. trachomatis were also determined. 1. Antigen detection from first-voided urine sediments of young adult men without urogenital symptoms (141 cases): The detection rate by IDEIA was 5.0% (7/141). Three of the 7 cases which were positive for antigen in first-voided urine sediments were reaffirmed as having asymptomatic C. trachomatis urethritis, since they were also revealed to have C. trachomatis in the urethra. 2. The positive rates of serum antibodies: The titers of serum antibody were determined in 128 cases out of the 141 cases. The positive rates of IgA and IgG were 6.3% (8/128) and 35.9% (46/128) respectively. The positive rates of IgA and IgG antibodies were significantly higher in cases with positive antigen in first-voided urine sediments than in those which were negative. These results indicate that this kit is useful for antigen detection. 3. The screening test revealed asymptomatic C. trachomatis infections in 5% of young adult men, suggesting the extensive spread of the infection. The screening test using first-voided urine sediments will be useful in public health.     

Diagnostic utility of estimation of mycobacterial antigen A60 specific immunoglobulins in serum and CSF in adult neurotuberculosis

An ELISA assay based on mycobacterial antigen A60 (Anda, Biologicals France) was used to detect specific immunoglobulins (IgM, IgA and IgG) in 48 cases of adult neurotuberculosis (24 TBM; 24 Tuberculoma) and in 48 controls (24 diseased controls; 24 healthy controls). Serum was analysed in all the subjects whereas CSF was assayed only in TBM cases and diseased controls. The cut off values used for IgM, IgG and IgA in this study were 1.500 ODI (optical density index) at 1:100 dil, 250 units/ml and 150units/ml respectively in serum; and 1.500 ODI at 1:10 dil, 10 units/ml and 10 units/ml respectively in CSF. The mean titres of all three antibodies were found to be significantly higher in cases as compared to controls. In cases of TBM, in serum, the percentage positivity for IgM, IgG, IgA and combination of IgG or IgA were 41.67, 87.50 87.50 and 95.83 respectively. The corresponding figures in CSF were 62.50, 75.0, 66.67 and 79.16 for IgM, IgG, IgA and 'IgA or IgM' respectively. In tuberculoma cases, in serum, the figures were 37.50, 75.0, 75.0 and 83.33 respectively. Overall, a high sensitivity and specificity were obtained in cases of TBM (Serum: ST = 95.83%: SP = 87.50%; CSF ST = 79.16%. SP = 100%) and Tuberculoma cases (serum: ST = 83.33% SP = 87.50%) employing the combined antibody estimations.     

Use of a peptide based enzyme immunoassay in diagnosis of Chlamydia trachomatis triggered reactive arthritis.

OBJECTIVE: To assess the presence of circulating IgA and IgG antibodies to Chlamydia trachomatis in sera of patients with reactive arthritis (ReA) and other arthritides. METHODS: A peptide based enzyme immunoassay (EIA) was used to study 132 patients divided into 5 groups: C. trachomatis triggered ReA, uroarthritis, enteroarthritis, oligoarthritis, and rheumatoid arthritis (RA). Followup sera were available from 19 patients. RESULTS: An increased prevalence of C. trachomatis antibodies was observed in patients with ReA triggered by C. trachomatis; 18/23 (78%) had IgA and 19/23 (83%) had IgG antibodies. In patient groups with uroarthritis (n = 12), enteroarthritis (n = 56), oligoarthritis (n = 16), and RA (n = 25), C. trachomatis IgA/IgG antibodies were detected in 58%/75%, 27%/21%, 25%/31%, and 20%/32% of patients, respectively. Both the IgA and IgG antibodies were positive in 74%, 50%, 16%, 25%, and 12% of the patients with C. trachomatis triggered ReA, uroarthritis, enteroarthritis, oligoarthritis, and RA, respectively. Based on positivity of both isotypes the sensitivity of the assay was 74% and specificity 84%. In the followup sera, an association between circulating C. trachomatis-specific antibody concentrations and clinical disease outcome of the arthritis was seen in patients with culture-positive C. trachomatis triggered ReA. CONCLUSION: C. trachomatis species-specific peptide EIA correlates well with conventional diagnosis of primary C. trachomatis infection in patients with ReA. This assay may be a valuable contribution to the diagnosis of C. trachomatis triggered ReA.     

Immunoblot analysis of antibody response to Chlamydia trachomatis in patients with reactive arthritis and ankylosing spondylitis

Antibodies to Chlamydia trachomatis were found in 60% of patients with reactive arthritis (ReA) and 33% of patients with ankylosing spondylitis (AS), compared with 19% of healthy blood donors. The IgG, IgA and IgM immune responses in patients with ReA and AS were further analysed by immunoblotting. Most patients had IgG antibodies to a large number of C. trachomatis antigens. IgA (and especially IgM) antibodies were less prevalent. Differences in the antibody response to individual antigens were seen between the two groups of patients, with respect to both IgG and IgA. Especially evident was the high prevalence of IgA antibodies to a 60 kD antigen among patients with ReA (67%) compared with patients with AS (20%).     

Evaluation of chlamydial IgM antibodies for clinical diagnosis]

Chlamydia trachomatis (C. trachomatis), C. psittaci, and C. pneumoniae are now well established as pathogens of respiratory infections including pneumonia. Serum samples from 223 infants and children with pneumonia, 31 patients with adult inclusion conjunctivitis, 16 parents of babies with neonatal inclusion conjunctivitis and others were tested for IgM antibodies to Chlamydiae. Diagnostic kits for chlamydial IgM antibodies (SeroELISA and IPAzyme) have been also evaluated for their diagnostic value. It was found that detection of specific IgM antibodies with SeroELISA has a diagnostic value in chlamydial pneumonias.     

The role of diagnostic tests in the identification of Chlamydia trachomatis infection in reactive arthritis

Reactive arthritis (ReA) is a sterile inflammation of the synovial membrane of one or more joints developing after urogenital or gastro-intestinal infection. The syndrome most frequently follows infection with Chlamydia trachomatis. Useful for the diagnosis can be the serological tests. At present there is the possibility to identify the specific antibodies (IgG, IgA, IgM) to Chlamydia trachomatis. The subject of the study was the group of 87 patients in age 19-78; 58 women and 29 men from whom urogenital smear and serum were tested. The control group were 30 people age 25-70 without rheumatological disorders. Chlamydia trachomatis was found in urogenital smear in 42 (48%), in 56 (64%) patients immunoglobulin IgG were positive, and immunoglobulin IgA in 16 (18%). The laboratory tests and clinical symptoms allow to make a diagnosis of ReA in 38 (43%) and possible ReA in 5 (5.7%) of patients. CONCLUSIONS: 1. There is no gold standard for the diagnosis of ReA. 2. None of the tests or the clinical symptoms alone are strong enough to make a definite diagnosis of ReA. 3. Tests to identify Chlamydia trachomatis, with respect of typical clinical symptoms are useful the diagnosis of ReA. 4. The diagnosis of ReA is most probable if we have typical clinical symptoms, clinical evidence of a preceding infection plus a positive result of serology or PCR plus positivity for HLA-B27.     

A study of secretory IgA antibody titers for Chlamydia trachomatis in prostatic secretion of chronic prostatitis

In order to study the correlation between chronic prostatitis and C. trachomatis, IgA and IgG antibody titers for C. trachomatis were measured in serum and prostatic secretion of cases of chronic prostatitis. IgA antibody titers have higher tendency in prostatic secretion than in serum. The other side, IgG antibody titers have higher tendency in serum than in prostatic secretion. This result suggested IgA antigen was reacted in local immunological response in prostatic gland. Subsequently, in order to confirm that IgA antibody in prostatic secretion is formed from secretory IgA antibody mainly, IgA antibody titers were compared with secretory IgA antibody titers. That results was considered that many cases with high IgA titers had high secretory IgA titers. The fact suggested in cases of C. trachomatis prostatitis, IgA antibody was mainly formed from secretory IgA antibody in prostatic secretion.     

Role of Chlamydia, mycoplasma and cytomegalovirus infection in the development of coronary artery disease

AIM: To assess relationship between some infection factors and presence of coronary heart disease. MATERIAL: Patients with myocardial infarction (n=56), unstable angina (n=50), stable angina (n=50) and age - matched controls (n=49). METHODS: Levels of IgG, IgM, IgA antibodies to Chlamydia pneumonia, Chlamydia trachomatis, Chlamydia psittaci, IgG, IgM antibodies to Cytomegalovirus, and also of antibodies and antigen to Mycoplasma pneumoniae were measured in blood serum. RESULTS: Compared with controls patients with coronary heart disease had higher frequency of seropositivity to Chlamydia pneumonia, Mycoplasma pneumonia and Cytomegalovirus (p< 0.05 ) and similar levels of seropositivity to Chlamydia trachomatis and Chlamydia psittaci. Infectious burden (quantity of antibodies per one patient) was significantly higher in patients with myocardial infarction, unstable and stable angina than in controls (1.58, 1.42, 1.41 and 0.95, respectively). CONCLUSION: Our results confirm presence of association between infection and coronary heart disease.     

Kinetic study of antibodies (IgG, IgA) to Chlamydia trachomatis: importance of IgA antibody in screening test for C. trachomatis infection by peptide-based enzyme immunosorbent assay

To assess the importance of only IgA antibody positivity in the peptide-based ELISA (P-ELISA) examination of kinetic behaviors of antibodies (IgA, IgG) to Chlamydia trachomatis, 426 sera from 52 follow-up antigen-positive patients were assayed. In part, a microimmunofluorescence (MIF) test and an immunoblot (IB) assay were also used for confirmation. The results showed that the positivity rates of IgA and IgG antibodies were 82.7 and 96.2%, respectively, at the first testing. One patient had both IgA- and IgG-negative antibodies at the first testing, but this became only IgA-positive and then IgG-positive. The patient was co-infected with Candida albicans and C. trachomatis, and saw a gynecologist for the symptom of itching. Although the major outer membrane protein was negative in IB assay, the results of the MIF test and absorption experiments were positive. MIF titers for IgA and IgG antibodies to C. pneumoniae were <1:8 and 1:32, respectively, at the peak level of P-ELISA. These findings seem to suggest that when only the IgA antibody is detected by P-ELISA, C. trachomatis infection may be present at an early stage, so confirmation via testing for C. trachomatis is needed.     

Recent trends in chlamydial and gonococcal conjunctivitis among neonates and adults in an Irish hospital.

BACKGROUND: Chlamydia trachomatis and Neisseria gonorrhoeae are two important and frequently overlooked causes of neonatal and adult conjunctivitis. OBJECTIVES AND METHODS: In order to improve primary treatment, prevention, and control of infection caused by these organisms, an analysis of all cases presenting from July 2002 to December 2006 at a major Irish regional teaching hospital was performed. RESULTS: There were 51 cases of conjunctivitis in total. Among neonates and adults, C. trachomatis was the most common cause of conjunctivitis. Of the adult patients, 75% were men. The annual incidence of adult chlamydial conjunctivitis increased yearly from 2002 and correlated with an overall increase in genital chlamydia infection in the region. Neonatal chlamydial conjunctivitis has an overall incidence of 0.65/1000 live births and is continuing to rise annually. In 2006, gonococcal conjunctivitis accounted for 20% of all cases of conjunctivitis caused by sexually transmitted bacteria presenting to our hospital. CONCLUSIONS: The recent increase in the incidence of gonococcal keratitis serves to remind us that this important infection should be borne in mind when treating cases of purulent conjunctivitis. The diagnosis of chlamydial and gonococcal conjunctivitis requires a high index of suspicion and prompt treatment with systemic antibiotics.     

Analysis of the monomeric IgA to polymeric IgA ratio in specific IgA antibodies for Chlamydia trachomatis--clinical studies in chlamydial urogenital infections

Antibody titers of monomeric and polymeric types of serum specific IgA for Chlamydia trachomatis were measured by indirect immunoperoxidase assay (Savyon kit) in male and female cases with various chlamydial urogenital infections. From these results, the ratio of monomeric to polymeric IgA (m/p ratio) was determined. All cases were positive for specific IgA and IgG antibodies for C. trachomatis, and the antigen was also detected in all of cases except for those with prostatitis. 1. Study in males: The m/p ratio (mean +/- S.E.) was 2.6 +/- 1.0 in acute chlamydial urethritis and 8.0 +/- 2.2 in chronic non-bacterial prostatitis. The result indicated monomeric IgA-predominance in the chronic stage. 2. Study in females: The m/p ratio was 5.0 +/- 1.9 in subacute chlamydial cervicitis, while it was 8.5 +/- 2.9 in pregnant women considered to have chronic chlamydial infection. The rate was 31.5 +/- 16.8 in prostitutes considered to have repeated chlamydial infections. The result suggested that monomeric IgA was predominant in patients with chronic and repeated infections. 3. As for sequential changes of IgA antibody titer, polymeric IgA alone decreased after treatment of acute chlamydial urethritis. However monomeric IgA decreased in chronic infection such as prostatitis, and a similar change was shown in IgG antibody. 4. These results suggest that polymeric IgA is predominant in the acute stage of chlamydial infections, while monomeric IgA predominates in the chronic stage.     

非典型病原菌致成人社区获得性肺炎患者的临床特点分析

目的 研究非典型病原菌感染在成人住院社区获得性肺炎中的重要地位,并对其临床特点进行分析.方法 收集2005年5月至2008年5月来自国内多中心的153例成人社区获得性肺炎住院患者急性期及恢复期双份血清和急性期痰标本,采用间接免疫荧光法检测肺炎衣原体IgG抗体及嗜肺军团菌IgG、IgA及IgM混合抗体滴度,采用被动凝集法检测肺炎支原体IgG、IgA及IgM混合抗体滴度,同时对153份急性期痰标本进行普通细菌培养.用回顾性分析方法了解非典型病原菌在成人社区获得性肺炎住院患者中的地位.结果 153例血清学检测结果中符合非典型病原菌致社区获得性肺炎诊断标准的52例(34.0%),其中47例为单一非典型病原菌感染,其中肺炎衣原体38例,肺炎支原体4例,嗜肺军团菌5例;5例为2种非典型病原菌混合感染,其中肺炎衣原体+肺炎支原体2例,肺炎衣原体+嗜肺军团菌3例;52例中合并细菌感染者11例.结论 非典型病原菌(肺炎衣原体、肺炎支原体及嗜肺军团菌)为成人住院社区获得性肺炎的重要致病菌,以肺炎衣原体为主,同时不能忽视合并细菌感染情况的存在.     

Serological evidence of Chlamydia pneumoniae lipopolysaccharide antibodies in atherosclerosis of various vascular regions

BACKGROUND: The role of Chlamydia pneumoniae in the pathogenesis of atherosclerosis has so far mainly been investigated in patients suffering from coronary heart disease; the other vascular regions have virtually been ignored. The aim of this study was to carry out a statistical survey of serological markers of a C. pneumoniae infection in patients with different patterns of atherosclerosis manifestation. PATIENTS AND METHODS: 340 patients were examined for the atherosclerotic alteration of peripheral arteries of the lower limbs, carotid arteries and coronary arteries by ultrasound scan and/or angiography. Immunoglobulin(Ig)G and IgA-rELISA were used to measure chlamydial lipopolysaccharide antibodies. Species determination was performed using the IgG micro-immunofluorescence test. RESULTS: 24.0% of atherosclerotic cases (A) and 52.3% of controls (C) were negative for C. pneumoniae lipopolysaccharide antibodies (p = 0.00002). By contrast, 45.1% of atherosclerotic cases and 16.9% of controls were positive for both IgG and IgA (p = 0.00002). The mean antibody titers of the atherosclerosis group were higher than in the control group (IgG positive xAIgG = 344, xCIgG = 272; IgG and IgA positive xAIgG = 576, xCIgG = 486 and xAIgA = 120, xCIgA = 91). Concerning atherosclerosis manifestation in various vascular regions, no significant differences were found between IgG and IgA antibody titers and prevalence. CONCLUSIONS: The results show that a persistent C, pneumoniae infection with evidence of lipopolysaccharide immunoglobulin G and A is equally associated with the atherosclerotic alteration of coronary arteries, carotid arteries and peripheral arterial occlusive disease, irrespective of the severity of atherosclerosis and with no predisposition to any particular vascular region.