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1.
An ELISA for detection of IgG, IgA, and IgM antibody using an acid-glycine extract from Campylobacter jejuni as antigen was developed. To determine the value of this assay for the diagnosis of acute Campylobacter jejuni infections, the IgG, IgA, and IgM immune response against Campylobacter jejuni was investigated at various timepoints after infection in patients with culture-proven infection. A total of 112 sera from 46 patients and 78 sera from a control group were tested. All but one of the 46 patients with culture-proven Campylobacter jejuni enteritis developed IgG antibodies against Campylobacter jejuni. IgA and IgM ELISA both showed 97% specificity, and sensitivity of 63% and 30% respectively. IgG antibody titers generally remained at a constant level for more than 50 days, whereas IgA and IgM antibody titers declined more rapidly to normal values within 30 to 50 days after onset of clinical symptoms. Detection of Campylobacter jejuni specific IgA antibodies in a single serum sample provided the most useful assay for serological diagnosis of Campylobacter jejuni enteritis. The presence of Campylobacter jejuni specific IgM antibodies was the sole diagnostic criterion in three cases. Serological diagnosis of Campylobacter jejuni enteritis should therefore include both IgA and IgM antibody determination.  相似文献   

2.
Chlamydia trachomatis was recoved from 20% (36/180) of women attending a venereal disease clinic. All infected women had chlamydial antibodies in their serum and cervical secretions. However, the background rates of chlamydial antibody in chlamydia-negative women were very high. Measurement of antibodies in serum (complement fixation or immunoglobulin G [IgG] and IgM by microimmunofluorescence) or cervical secretion (IgG, IgM, IgA or secretory IgA classes) did not result in predictive values of greater than 32%. It is concluded that the detection of chlamydial antibodies in serum or cervical secretions cannot be substituted for agent isolation in diagnosing these infections.  相似文献   

3.
Cytomegalovirus (CMV)-specific IgA antibodies were determined by an immunoperoxidase assay in sequential serum samples of 10 patients with CMV infection in order to evaluate the feasibility of the use of this technique for diagnosis. In parallel, IgM and IgG antibodies to CMV were studied by enzyme-linked immunosorbent assay (ELISA) and by the immunoperoxidase assay, respectively. CMV IgA antibodies were detected in all 10 CMV patients studied. Specific IgM was detected earlier than IgA in only one of these ten patients. No CMV-specific IgA antibodies (titer less than 2) were detected in 45 medical students. Neither were they found in paired sera of 5 patients with herpes simplex infection, 5 patients with varicella, 6 patients with zoster and 2 patients with Epstein-Barr virus infection. The potential application of the indirect immunoperoxidase IgA assay for serodiagnosis of CMV infections is discussed.  相似文献   

4.
Enterovirus A71 (EV-A71) is one of the main pathogens causing hand, foot, and mouth disease, and often causes diseases of the central nervous system. Early diagnosis is important to prevent EV-A71 outbreaks. The detection of serum immunoglobulin M (IgM) is widely used for the early diagnosis of EV-A71 in clinics, especially in rural areas. However, this technique requires the extraction of blood from children who have thin blood vessels and who might fear the use of needles. Therefore, difficulties in the detection process are often encountered. This study developed a noninvasive method to detect EV-A71-specific immunoglobulin A (IgA) in saliva for the diagnosis of EV-A71 infection. The sensitivity and specificity of IgA detection did not differ significantly compared with IgM detection. IgA antibodies were present in saliva for a relatively shorter period than IgM antibodies were present in serum. The sensitivity of IgA detection was higher than that of IgM detection for secondary EV-A71 infections. These results suggest that the detection of EV-A71-specific IgA in the saliva allows the effective early diagnosis of EV-A71 and may be suitable for detecting EV-A71 infections in children.  相似文献   

5.
An enterovirus-specific indirect ELISA, based on a single local isolate of coxsackie B5 as antigen, was used to study the IgA, IgG, and IgM responses in 19 patients with a recent or current enterovirus infection. Twelve different enterovirus serotypes were isolated from 15 patients. Paired serum samples were available from 10 and a single serum from 5 of these 15 patients. In addition, 4 patients diagnosed by a significant titer rise of complement fixing antibodies to enterovirus were included. A serological diagnosis, defined as an increase in titer of enterovirus IgG and/or presence of enterovirus IgM, were established in all 14 patients with paired sera. Enterovirus IgM was present in either a single serum or in both sera in 13 of them. Out of 5 patients with a single serum sample only, enterovirus IgA or enterovirus IgM was found in 4. Specific IgA was present in either a single serum or in both sera in 14 of the 19 patients. Seven of the 10 enterovirus isolate patients with paired sera had a significant titer rise of complement fixing antibodies; however, all 10 were diagnosed by ELISA. Among 64 healthy controls 2 had enterovirus IgA and none had enterovirus IgM. In conclusion, the use of a single antigen-based ELISA was found to be reliable for the diagnosis of recent and current enterovirus infections.  相似文献   

6.
To evaluate alternative approaches to the serological diagnosis of dengue virus (DEN) infection, the detection of DEN-specific immunoglobulin M (IgM) and IgA antibodies in serum and saliva specimens was assessed in 147 patients with symptoms of DEN infection seen at the Ministry of Health in Nicaragua. Seventy-two serum samples were determined to be positive for anti-DEN antibodies by IgM capture enzyme-linked immunosorbent assay, the routine diagnostic procedure. Serum and saliva specimens were obtained from 50 healthy adults as additional controls. IgM was detected in the saliva of 65 of the 72 serum IgM-positive cases, 6 of the 75 serum IgM-negative cases, and none of the control group, resulting in a sensitivity of 90.3% and a specificity of 92.0% and demonstrating that salivary IgM is a useful diagnostic marker for DEN infection. Detection of IgA in serum may be another feasible alternative for the diagnosis of DEN infection, with serum IgA found in 68 (94.4%) of the IgM-positive cases. In contrast, detection of IgA in saliva was not found to be a useful tool for DEN diagnosis in the present study. Further studies of the kinetics of antibody detection in another set of 151 paired acute- and convalescent-phase serum samples showed that DEN-specific IgA antibodies were detected in more acute-phase samples than were IgM antibodies. Thus, we conclude that DEN-specific IgA in serum is a potential diagnostic target. Furthermore, given that saliva is a readily obtainable, noninvasive specimen, detection of DEN-specific salivary IgM should be considered a useful, cheaper diagnostic modality with similar sensitivity and specificity to IgM detection in serum.  相似文献   

7.
To evaluate alternative approaches to the serological diagnosis of dengue virus (DEN) infection, the detection of DEN-specific immunoglobulin M (IgM) and IgA antibodies in serum and saliva specimens was assessed in 147 patients with symptoms of DEN infection seen at the Ministry of Health in Nicaragua. Seventy-two serum samples were determined to be positive for anti-DEN antibodies by IgM capture enzyme-linked immunosorbent assay, the routine diagnostic procedure. Serum and saliva specimens were obtained from 50 healthy adults as additional controls. IgM was detected in the saliva of 65 of the 72 serum IgM-positive cases, 6 of the 75 serum IgM-negative cases, and none of the control group, resulting in a sensitivity of 90.3% and a specificity of 92.0% and demonstrating that salivary IgM is a useful diagnostic marker for DEN infection. Detection of IgA in serum may be another feasible alternative for the diagnosis of DEN infection, with serum IgA found in 68 (94.4%) of the IgM-positive cases. In contrast, detection of IgA in saliva was not found to be a useful tool for DEN diagnosis in the present study. Further studies of the kinetics of antibody detection in another set of 151 paired acute- and convalescent-phase serum samples showed that DEN-specific IgA antibodies were detected in more acute-phase samples than were IgM antibodies. Thus, we conclude that DEN-specific IgA in serum is a potential diagnostic target. Furthermore, given that saliva is a readily obtainable, noninvasive specimen, detection of DEN-specific salivary IgM should be considered a useful, cheaper diagnostic modality with similar sensitivity and specificity to IgM detection in serum.  相似文献   

8.
Twenty-three patients with a herpetic infection as diagnosed by a positive culture of herpes simplex virus (HSV) were studied with respect to serological responses of IgA, IgG and IgM antibodies in paired serum samples by an indirect (sandwich) enzyme linked immunosorbent assay (ELISA). Eight of the patients had a primary infection and 15 a recurrent one. In the ELISA test a detergent treated cell lysate of HSV type 1 was used as antigen. In the IgM assay all sera were pretreated with antihuman IgG with the purpose to precipitate IgG of the samples. The conjugate was a F(ab)2-fragment of antihuman-IgM. In primary infections all patients had significant titre rises of IgG and presence of high IgM titres in the convalescent serum. IgA antibodies were found in all of them, while titre rises were detected in 5/8. In recurrent infections titre rises of IgG and IgA antibodies were found in 4 and 5, respectively. Six had detectable IgM in one or both of the paired samples. The IgG titres were higher in recurrent infections than in primary, in contrast to IgM of which much higher titres were found in primary infections. It is concluded that in primary infections a conclusive serological diagnosis was established in all patients, whereas in recurrent infections this was achieved in two of three patients. The indirect ELISA method used for IgM detection was sensitive, reliable and convenient. Interfering rheumatoid factor was effectively eliminated by treatment with antihuman IgG.  相似文献   

9.
The microimmunofluorescence technique (MIF) is recognized as the only test hitherto allowing discrimination between different Chlamydia species and is considered to be the reference method for serology. This method was developed for the detection of IgG and IgM antibodies only. We investigated the effects of some test parameters on the ability of MIF to detect Chlamydia pneumoniae IgA. These parameters were the time needed for binding of serum IgA to C. pneumoniae antigen and the effect of antigen concentration on the outcome of IgA antibody testing. It was found that the most sensitive MIF tests for the detection of serum IgA antibodies were those in which an overnight incubation of sera with antigen slides containing high concentrations of chlamydial elementary bodies was employed. The number of patients with chronic infections found to have elevated IgA titers was increased by 25% using longer incubation times for the antibody-antigen reaction. Thirty-two sera from patients with coronary artery disease and confirmed chronic C. pneumoniae infection were used to compare antigen slides with low and high concentrations of elementary bodies with respect to IgA levels; 31/32 patients were found to have specific IgA antibodies to C. pneumoniae using the high antigen concentration, as opposed to only 22/32 patients using the low antigen concentration.  相似文献   

10.
Sera from 35 patients with acute pelvic inflammatory disease (PID) with and without Chlamydia trachomatis confirmed by culture and sera from 19 control patients with neither evidence of pelvic infection nor C trachomatis infection were studied for the presence of serum IgG, IgA, and IgM antibodies to C trachomatis using enzyme immunoassay (EIA) and immunoblotting techniques. There was no correlation between the antibody concentrations in the EIA and the spread of chlamydial infection, as determined by cervical, endometrial, and laparoscopic sampling for chlamydia. The immunoblot analysis showed antibodies to the major outer membrane protein (MOMP) of C trachomatis elementary bodies in all patients who had had C trachomatis isolated. Reactivity was also frequently observed against the 68, 62, 60, 45, and 31 kilodalton antigens. About 20 antigenic polypeptides were identified. Differences in antibody prevalence to specific chlamydial antigens, however, were not related to the site of chlamydial isolation or serum antibody concentrations observed with the EIA. The results indicate that patients with PID with and without upper genital tract infection with C trachomatis cannot be differentiated by reactivity of sera to specific chlamydial polypeptide antigens. The determination of a specific serum IgA antibody response by EIA was the most effective single test to discriminate between patients with and without acute chlamydial infection.  相似文献   

11.
12.
Herpes simplex virus (HSV)-specific IgG and IgA antibody response in patients with HSV infection and in healthy adults was studied by the immunoperoxidase antibody-membrane antigen (IPAMA) technique. In all HSV infections in which specific IgM antibodies were detected by enzyme-linked immunosorbent assay (ELISA), a significant rise in the titer of HSV IgG and IgA antibodies was found. In contrast, in patients with recurrent herpes labialis in which no specific HSV IgM antibodies were detected by ELISA, HSV IgG and IgA antibodies were not found to fluctuate significantly during the course of infection. A higher geometric mean titer (GMT) for HSV IgG and for IgA antibodies was found in seropositive individuals with a previous history of recurrent HSV than in seropositive individuals without a previous history of recurrent HSV infection. Nineteen of 26 HSV IgG seropositive healthy medical students without a previous history of recurrent HSV infection had HSV IgA antibodies to membrane antigen. The significance of this finding in understanding the mechanism of latency in healthy seropositive individuals without previous history of HSV recurrent infections is discussed.  相似文献   

13.
INTRODUCTION: The aim of the study was to evaluate the prevalence of anti-Chlamydia trachomatis (C.trachomatis) antibodies in serum and expressed prostatic secretion (EPS) in chronic prostatitis. MATERIALS AND METHODS: Thirty-six patients with chronic prostatitis were examined. The presence of C. trachomatis was determined in urethral smears and EPS. Specific antibodies were determined in the serum (IgM, IgA, IgG) and in the EPS (IgA, IgG). In the direct diagnosis of chlamydial infection, the direct immunofluorescence method and the ligase chain reaction were employed, and for the serological diagnosis, the immunoenzymatic method. RESULTS: C. trachomatis infection was detected in the urethra of 3 (8.3%) patients and in the prostatic gland of 3 (8.3%) patients; only one of these patients was found to have C. trachomatis in both the urethra and the EPS. In the control group, C. trachomatis was detected in the urethra of 1/50 (2%) of the men, but the EPS of all of them was free of C. trachomatis. Specific IgM antibodies were found in 7 (19.4%), IgA in 9 (25%), and IgG in 18 (50%) of the patients' serum, whereas IgAs were detected in 12 (33.3%) and IgGs in 13 (36.1%) of the patients' EPS. In the control group, anti-C. trachomatis antibodies of the IgG were detected in the serum of 2/35 (5.7%) of the men, whereas in the EPS neither IgA nor IgG antibodies were detected in any of these patients. CONCLUSIONS: Serological tests of the serum and EPS are useful as a complementary method in the diagnosis of chronic prostatitis.  相似文献   

14.
Most of the studies on 60-kDa and 10-kDa chlamydial heat shock proteins (HSPs) to date have been carried out with blood lymphocytes or serum antibody responses, which do not provide a clear picture of the actual pathogenesis as they do not differentiate primary infection from recurrent infection. Thus, in the present study induction of the immune response was evaluated by studying lymphoproliferation of both cervical and peripheral lymphocytes to synthetic peptides of cHSP60, cHSP10 and major outer membrane protein (MOMP) antigen. In addition, cervical antibody prevalence to MOMP antigen, cHSP60 and cHSP10 and cytokine levels in cervical washes was also determined. Positive proliferative responses of cervical lymphocytes to cHSP10 peptide were significantly higher (P < 0.05) in women with recurrent infections and that to MOMP antigen were significantly higher in primary infection. On proliferation of PBMCs with the above antigens, no significant difference was observed between primary and recurrent infection. Prevalence of cervical IgG and IgA antibodies to Chlamydia trachomatis was significantly higher (P < 0.05) during primary infection than recurrent infections. In contrast, prevalence of IgG and IgA antibodies to cHSP10 and IgG antibodies to cHSP60 was higher during recurrent infections than primary infections. Interferon (IFN)-gamma levels were significantly higher in cervical washes of women with recurrent infection and correlated strongly with cHSP60 antibody titres. Our data thus suggest that mucosal responses are more appropriate in understanding the pathogenesis of chlamydial infection and IFN-gamma could be involved in the modulation of immune responses towards chlamydial infection directly, by causing acute inflammation, or indirectly through modulation of HSP expression.  相似文献   

15.
Toxoplasma gondii specific IgA and IgM antibodies were quantitated by an antibody capture agglutination assay in 260 patients with acquired toxoplasmosis and from 94 fetuses suspected of congenital toxoplasmosis and 30 infected children. In acquired toxoplasmosis, IgA antibodies to T gondii were found in 95% of the cases. In congenital toxoplasmosis IgA antibodies were more frequently detected (75%) in cord blood than IgM antibodies (61%). They persisted after birth, in some cases for up to 24 months. IgA antibodies were also detected in fetuses whose mothers had toxoplasmosis during their pregnancy. In infected fetuses IgM and IgA antibodies were detected in fetal blood as early as week 24 of pregnancy. Detection of IgA T gondii antibodies may be useful for the diagnosis of some recently acquired infection and for the diagnosis and follow up of the infection in the fetus and neonate.  相似文献   

16.
The objective of the present study was to evaluate the clinical usefulness of the simultaneous measurement of three serological markers of chlamydial infection in women with tubal factor infertility (TFI) and spontaneous miscarriage. Serum was collected from 87 patients (33 with TFI and 54 with spontaneous miscarriage) and analyzed for the presence of IgG and IgA antibodies against Chlamydia trachomatis MOMP antigen (Dia.Pro) and IgG antibodies to chlamydial heat shock protein 60 (cHSP60) antigen (Medac). We determined a high degree (64.5 %) of seropositivity against chlamydial antigens in our study population. The prevalence of persistent chlamydial infection has tended to be higher in the group of patients with TFI (41.4 %) than in patients with spontaneous miscarriage (21.3 %). The serum level of IgA, as a marker of active infection, was statistically higher in the TFI group with persistent infection than in the corresponding spontaneous miscarriage group (p?=?0.008), while the serum level of IgG showed no statistically significant differences compared with the spontaneous miscarriage group with persistent infection (p?=?0.227). Also, using the receiver operating characteristic (ROC) curve, we found that the serum level of IgA has the ability to discriminate patients with persistent chlamydial infection between the TFI and miscarriage groups, with a sensitivity and specificity of 74.3 % and 71.4 %, respectively. To the best of our knowledge, the present study is the first study which, besides the already confirmed linkage between serologic evidence of persistent chlamydial infection and TFI, also confirmed associations between spontaneous miscarriage and serologic evidence of persistent chlamydial infection.  相似文献   

17.
Data published over the past decade show that Chlamydia pneumoniae is likely associated with the development of atherosclerosis. The aim of this study was to ascertain whether C. pneumoniae infections occur more frequently in patients with atherosclerosis than in healthy subjects. A total of 517 persons were studied. Serum samples, leukocytes, and tissue samples were assayed for the presence of C. pneumoniae-specific IgG and IgA antibodies and C. pneumoniae DNA. C. pneumoniae DNA was found in renal, iliac, and brachial vessels, but it was not detected in radial arteries. C. pneumoniae DNA was found most often in directional coronary atherectomy tissue specimens (11/41, 26.8%), but it was also found in the leukocytes of 14.9% (28/188) of patients with atherosclerosis and 24.6% (28/114) of patients without atheroma changes in vessels. Specific IgG and IgA antibodies were present in 63.8 and 49.9% of atheroma patients, respectively. The prevalence of C. pneumoniae antibodies differs significantly in patients with and without atherosclerosis (for IgG, p=0.002, and for IgA, p=0.006). The identification of persons with chlamydial infection of atherosclerotic arteries necessitates the examination of vascular tissues obtained during revascularization procedures. Serological investigation alone cannot identify individuals with vascular chlamydial infections. Detection of C. pneumoniae DNA in peripheral blood mononuclear cells does not seem to be the exclusive marker of persistent vascular infection. A more easily accessible parameter that allows prediction of chlamydial vascular infection is required.  相似文献   

18.
The detection of specific IgM antibodies has been the most frequently used serological marker for diagnosing recent toxoplasmosis. However, the persistence of specific IgM antibodies in some patients and the use of tests with a low specificity have complicated the interpretation of serological results when toxoplasmosis is suspected. The purpose of the present study was to determine the value of newer serological techniques in the diagnosis of acute acquired toxoplasmosis. Sixty-four sera, 31 from patients with Toxoplasma gondii infection and 33 from patients with latent infection, were tested. Anti-T. gondii IgA was measured by two antibody capture ELISA tests (Platelia Toxo IgA and ETI-TOXOK A) and an automated direct ELISA (IMx Toxo IgA); all three assays detected antibody levels compatible with a recent infection in sera from all 31 patients with acute toxoplasmosis. However, significant levels of IgA were also detected with high frequency by all three assays in sera from patients with latent infection. IgE antibodies detected by IgE immunosorbent agglutination assay (ISAGA) were present in 26 (84%) of 31 patients with acute toxoplasmosis and in sera from two subjects with latent infection taken >1 year after the beginning of the clinical symptoms of infection. Thirty (97%) of 31 patients with a recent T. gondii infection and 15 (45%) of 33 subjects with latent infection had an AC/HS pattern compatible with acute toxoplasmosis. The avidity of T. gondii IgG was evaluated by two methods. One method was based on the titration of each serum sample and calculation of the titres, in the absence and presence of urea, in relation to a defined cut-off value. In the other method, a single serum dilution was used and the absorbances of the reactions in the presence and absence of urea were compared. The titration method was more sensitive for diagnosing recent primary infection; all 31 sera from patients with acute toxoplasmosis had avidity indices compatible with acute toxoplasmosis by the titration method, whereas with the single dilution method, sera from four patients had equivocal results. In the 33 individuals with latent infection, similar results were obtained with the two avidity methods; only one serum sample had a non-compatible avidity value with the titration method. The results obtained in the present study show that the current serological markers used for diagnosing acute acquired toxoplasmosis have significant limitations. The data suggest that determination of the avidity of T. gondii-specific IgG by the titration method in patients with detectable IgM antibodies defines most accurately the stage of infection by T. gondii.  相似文献   

19.
Immunoglobulin class-specific enzyme immunoassay (EIA) was used for determination of antibody responses in sera collected from 26 children with acute primary respiratory syncytial virus (RSV) infections. All 26 patients had IgG antibody responses with a significant titer increase in 24 (92%); an IgM antibody response was detected in 19 of the 26 (73%). From patients aged 6 months or less only 5 of 8 produced detectable IgM antibodies, whereas all patients aged 1–2 years did so. IgM antibodies appeared within 1 week after onset of illness and persisted from 20 days to 2–3 months. An IgA antibody response was observed in 20 of 26 (77%) patients with a significant titer increase detected in 17 of 26 (65%) patients. In some patients the persistence of IgA antibodies followed that of the IgM antibodies, but in others the IgA antibody titers remained stable up to the end of the follow-up. The most sensitive assay system for serological diagnosis of acute RSV infection in children was the determination of titer increases by IgG antibody.  相似文献   

20.
Immunoblot and an enzyme-linked immunosorbent assays were used to evaluate and compare IgG antibodies against S. aureus whole cell lysate, cell wall peptidoglycan and lipoteichoic acid to discriminate between deep-seated and superficial S. aureus infection. Serum samples were examined from patients with deep-seated (n = 25) and superficial (n = 25) S. aureus infections and 15 healthy controls. Patients with deep-seated infections exhibited a large number of immuno-reactive bands in their IgG immunoblot profile as compared to those with superficial infections and healthy controls. Anti-staphylococcal IgG antibodies that reacted with two antigens of apparent molecular weight 110 and 98 kDa were specifically present in 96% (24/25) of patients with deep-seated infections, and were absent in, superficial and control sera. Moreover other Gram-positive and Gram-negative bacteria did not share these two unique antigens. The ELISA assays revealed significantly elevated levels of IgG antibodies to peptidoglycan (PG) in 18 of 25 (72%) patients with deep infection and 15 of 25 (60%) patients with superficial staphylococcal infection. The elevated levels of IgG antibodies to teichoic acid (TA) antigen were detected in all (100%) deep-seated group patients and among 40% (10/25) patients with superficial infection. An increase in levels of antibodies to PG showed a positive correlation trend with levels of IgG antibodies to TA only in deep infection group. Thus immunoblot detection of these two unique antigens as well as detection of elevated antibodies against PG and TA may be useful for the discrimination of staphylococcal deep-seated and superficial infection in humans.  相似文献   

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