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A simplified serological test for leprosy based on a 3,6-di-O-methylglucose-containing synthetic antigen.

The recent advent of synthetic antigens containing the Mycobacterium leprae-specific epitope, 3,6-di-O-methyl-beta-D-glucopyranoside, has allowed the development of simple specific serological tests for leprosy. The incorporation of one such product, 8-carbonyloctyl O-[4-O-(3,6-di-O-methyl-beta-D-glucopyranosyl)-alpha-L- rhamnopyranoside]-BSA, into a simple "spot" test, diffusion-in-gel enzyme-linked immunosorbent assay (ELISA), allowed an over 90% detection rate of untreated lepromatous leprosy, and the results showed good concordance with conventional ELISA based on the native phenolic glycolipid I.

Antigens, Bacterial↗

[Evaluation of the sensitivity of selected serological tests and activity of Coxiella burnetii antigens in the diagnosis of Q fever].

Sensitivity of three serological tests: indirect immunofluorescence assay (If), complement fixation test (CF), and microagglutination test (MA) was evaluated. Sera (118 samples) of humans suspected of C. burnetii infection were tested. Phase II antibodies were detected in 68.6% of sera and phase I antibodies--in 38.2% of sera. Among seropositive to phase II antigen--93.8% of sera reacted in IF, 62.9% in MA, and 32.1% in CF; among seropositive to phase I antigens--100% of samples reacted in IF, 2.6% in MA and 2.6% in CF. Calculated sensitivity of above tests was as followed: IF-93.8%, MA-67.1%, CF-34.2%. Some human sera (6.1%) reacted with hen egg antigens in CF. Reactivity of diagnostic antigens prepared from reference Henzerling strain and four others isolated in Poland with rabbit immune sera and sera of individuals suspected of C. burnetii infection in IF was compared. Generally, the immune sera reacted in highest titres with homologous antigens derived from homologous strains. Human sera showed differentiated activity to particular antigens. The titres of phase I antibodies fluctuated from 0 to 16 depending on the antigen applied. Because of that fact diagnostic antigens should be prepared from the mixture of reference strains and isolates from a region under study.

Agglutination Tests↗

Comparison of serological tests for detection of Mycoplasma hominis antibodies in female Grivet monkeys with experimentally induced salpingitis.

Four serological tests, viz. indirect hemagglutination (IHA), metabolism inhibition (MI), immunofluorescence (IMF), and a modification of the growth-inhibition (GI) test have been compared for the detection of antibodies against Mycoplasma hominis in female grivet monkeys with experimentally induced pelvic inflammatory disease. Moreover, cold hemagglutinins (CHA), and immunoglobulins, M, G, and A have been determined. The IHA test was found to be superior to the other methods used. The antibodies were present in all inoculated monkeys two weeks after the inoculation and a maximum in titre occurred one week later. Antibodies detected by the modified GI test occurred in all monkeys inoculated with the organism together with the IHA antibodies, but the maximum in titre was less marked. The IMF test was less sensitive than the GI test; the antibodies generally occurred later and disappeared faster. Using the MI test, no antibodies could be detected in any of the monkeys during the experimental period and no CHA antibodies were found in any of the sera. IgM rose earlier and declined more rapidly than IgG. It is concluded that the IHA test is most suitable for the measurement of serum antibodies caused by infection with M. hominis in grivet monkeys. The modified GI test, although less sensitive, may also be useful because of its simplicity in performance.

Animals↗

Comparative evaluation of rheumatic activity. A study of relationship between histologic changes and serologic test results in cardiac surgical patients.

Atrial or ventricular myocardium from patients with surgically corrected rheumatic valvular disease was studied for rheumatic lesions in 86 cases. Histologically active Aschoff bodies were found in 20 per cent of the cases. A slight, but statistically not significant relationship was demonstrated in comparison of elevated serologic tests for rheumatic activity with the presence of Aschoff bodies.

Adolescent↗

Use of frozen neutrophils in serological testing.

We describe an immunofluorescence test to detect neutrophil-specific antibodies using neutrophils fixed to a slide and frozen prior to use. In titration studies using NA1, NA2 and NB1 antisera the technique was less sensitive than the standard immunofluorescence test but did enable slides to be stored for at least 5 months with no loss of NA1, NA2 and NB1 antigens. This technique can be used as a simple alternative to neutrophil cytoplasts for the identification of neutrophil antibodies. However, its use as a screening test in neutrophil serology is limited by the occurrence of false negative reactions with weakly reactive antisera.

Antibodies↗

Establishment of the serologic testing algorithm for recent human immunodeficiency virus (HIV) seroconversion (STARHS) strategy in the city of São Paulo, Brazil.

Several strategies aim at characterizing the AIDS epidemic in different parts of the world. Among these, the identification of recent HIV-1 infections using the recently described serologic testing algorithm for recent human immunodeficiency virus (HIV) seroconversion (STARHS) strategy was employed in four testing sites of the City of Sao Paulo Public Health Department (CSPPHD). Those identified as recently infected were invited to participate in a prospective clinical and laboratory evaluation study. We describe the establishment of the patient identification network and the success in enrolling the participants, as well as their clinical and laboratory characteristics. From May to December 2002, 6,443 persons were tested for HIV in the four participating sites, of whom 384 (5.96%) tested HIV-1 positive; 43 (11.2%) of them were identified as recently infected. Twenty-two were successfully enrolled in the follow-up study, but three of them did not meet clinical and/or laboratory criteria for recent HIV-1 infection. After these exclusions, the laboratory findings revealed a median CD4+ T lymphocyte count of 585 cells/microL (inter-quartile range 25-75% [IQR], 372-754), a CD8+ T lymphocyte count of 886 cells/microL (IQR, 553-1098), a viral load of 11,000 HIV-RNA copies/mL (IQR, 3,650-78,150), log10 of 4.04 (IQR 3.56-4.88). The identification of recent HIV infections is an extremely valuable way to evaluate the spread of the virus in a given population, especially when cohort studies, considered the gold standard method to evaluate incidence, are not available. This work demonstrated that establishing a network to identify such patients is a feasible task, even considering the difficulties in a large, resource-limited country or city.

Adult↗

Results of a new serologic test suggest an association of Waddlia chondrophila with bovine abortion.

Waddlia chondrophila is a little-known intracellular organism belonging to the order Chlamydiales that has twice been isolated from aborted bovine fetuses. To initiate an investigation of the possibility that W. chondrophila may be an abortifacient pathogen, a serologic test was developed and used to screen bovine sera that were submitted to the Washington Animal Disease Diagnostic Laboratory (Pullman, WA). A highly significant statistical association (P < 0.00001) was observed when comparing antibody titers in cows that had aborted with those in other classes of cattle. Although this result is consistent with the possibility that infection with W. chondrophila may be a cause of bovine abortion, it is also possible that seroprevalence simply increases with age or that exposure rates differ between cows and other classes of cattle. Future serologic studies should specifically compare antibody titers between aborting cows and carefully matched nonaborting cohorts.

Abortion, Veterinary↗

Serological testing in malaria.

The main purpose of this paper is to evaluate, in a critical manner, various serological tests with general emphasis on their value in the epidemiological assessment of malaria. Several tests have been employed in the past. However, the present memorandum will deal only with the methods that have been widely used recently-i.e., indirect immunofluorescence (IFA), passive haemagglutination (IHA), and gel-diffusion. The three immunoglobulins most commonly involved in these tests are IgG, IgM, and-to a lesser extent-IgA.

Animals↗

Evaluation of cerebrospinal fluid in Southeast Asian refugees with reactive serologic tests for syphilis.

To determine the prevalence of cerebrospinal fluid abnormalities in Southeast Asian refugees with reactive serologic tests for syphilis, we evaluated 65 patients, 36 prospectively and 29 retrospectively, in a primary care clinic. Information was collected on history of treponemal infections, neurologic symptoms and signs, and total protein concentration, leukocyte count, and the VDRL test in the cerebrospinal fluid. Neurologic symptoms were reported by all patients for whom data were available. Abnormal neurologic signs were found or noted in medical records in 15 (42%) prospectively evaluated patients and 9 (64%) of 14 retrospectively evaluated patients for whom data were available. No patient had evidence of congenital or non-neurologic sequelae such as cutaneous or cardiovascular manifestations of syphilis. No patient had a positive cerebrospinal fluid VDRL test, 1 had more than 5 x 10(6) leukocytes per liter (5 leukocytes per mm3), and 6 (9%) had elevated total protein levels in the cerebrospinal fluid. Previous therapy for syphilis was not associated with lower serum VDRL reactions, neurologic symptoms and signs, or cerebrospinal fluid findings. In the absence of other indications, routine examination of the cerebrospinal fluid in seropositive Southeast Asian refugees who have nonspecific neurologic symptoms has a low yield, perhaps because of the high prevalence of yaws in this population, and may not be warranted.

Adolescent↗

Pattern of reactive serological tests for syphilis in different population groups attending the University College Hospital Ibadan (1976-1985).

A review of the reactive serological tests for syphilis was carried out. An overall seroreactivity of 5.1% was obtained over the 10-year period reviewed. A subtle but steady rise in the incidence of syphilitic infections was noted, from 7 to 10 new cases per 1000 population between 1970 and 1975. The predictive characteristics obtained from the screening tests show that the tests are reliable diagnostic tests when the results are carefully interpreted, and the medical personnel can be alerted of the prevalence of syphilitic infections.

Hemagglutination Tests↗

Differences in the temporal trends of HIV seroincidence and seroprevalence among sexually transmitted disease clinic patients, 1989-1998: application of the serologic testing algorithm for recent HIV seroconversion.

The authors compared temporal trends in the prevalence and incidence of human immunodeficiency virus (HIV) infection based upon 34,866 specimens from patients who attended the San Francisco, California, municipal sexually transmitted disease clinic between 1989 and 1998. HIV infection data were collected during annual blinded HIV serologic surveys. Incidence was determined by applying a serologic testing algorithm for recent HIV seroconversion that uses both a sensitive and a less sensitive enzyme immunoassay to stored HIV positive sera. The HIV seroprevalence declined from 15.2% in 1989 to 7.2% in 1998 (odds ratio per year = 0.92, 95% confidence interval (CI): 0.91, 0.94). Among homosexual men, the HIV prevalence declined from 50.9% in 1989 to 19.9% in 1998 (odds ratio per year = 0.86, 95% CI: 0.85, 0.88). The pooled seroincidence was 1.6% and did not change significantly over time (odds ratio per year = 1.0, 95% CI: 0.98, 1.1). The pooled seroincidence among homosexual men was 6.6% per year and remained steady between 1989 and 1998 (odds ratio per year = 0.99, 95% CI: 0.92, 1.1). During a dramatic, 10-year decline in seroprevalence of HIV infection, the incidence of HIV infection remained remarkably stable.

Adult↗

Diagnosis of Johne's disease in cattle: a comparison of five serological tests under field conditions.

Sera from 775 adult milking cattle on 13 farms in Friesland and from 359 of the same cattle taken during the following year, were examined for evidence of infection with M johnei by the complement fixation, fluorescent antibody, haemagglutination, haemagglutination-lysis and immunodiffusion tests. Eighteen animals, not clinically affected but which had serologically positive tests were slaughtered and M johnei was isolated in cultures from 16. The fluorescent antibody test was as sensitive as the complement fixation test. The other tests were relatively insensitive and would only be of value as confirmatory tests in the presence of reactions to the first two tests. It is concluded that the use of the fluorescent antibody and the complement fixation test together was of value in detecting sub-clinical Mycobacterium johnei infection.

Animals↗

Dark ground microscopy and treponemal serological tests in the diagnosis of early syphilis.

OBJECTIVES: To evaluate the use of dark ground microscopy (DGM) and treponemal serological tests in the diagnosis of primary (PS) and secondary (SS) syphilis. METHODS: A retrospective case note review of patients with early syphilis who attended our department between January 2001 and December 2002. Data were collected on demographics, results of treponemal serology and DGM. RESULTS: 50 individuals had PS and 36 individuals had SS. DGM was performed in 31/50 (62%) of PS cases and this was positive in 97%. In 17 (34%) cases of PS, treponemal EIA was negative initially. DGM was performed on 13 of these, all of which were positive. Therefore, EIA had a sensitivity of 57% when compared to DGM. In 27 patients where EIA-IgM was performed, this was positive in 22 (81%), of which 12 were EIA negative on initial screening. All SS cases had positive EIA. DGM was performed in 19/36 (52%) of SS cases and was positive in 16/19--that is, a sensitivity of 84% when compared to EIA. The major reason why DGM was not performed in the cases of PS was that herpes was the presumed diagnosis and in SS the rash was attributed to other causes. CONCLUSIONS: DGM is a rapid and sensitive test while EIA takes time for results and is less sensitive in PS. EIA-IgM is a useful adjunct in PS. DGM allows immediate diagnosis, treatment, and partner notification preventing further transmission. Genitourinary medicine clinics should have trained staff to perform DGM on all anogenital ulcers and suspected syphilitic lesions.

Cohort Studies↗

[Prevalence of American trypanosomiasis, syphilis, toxoplasmosis, rubella, hepatitis B, hepatitis C, human immunodeficiency virus infection, assayed through serological tests among pregnant patients, from 1996 to 1998, at the Regional University Hospital Norte do Paraná].

In order to evaluate the seroprevalence of the american trypanosomiasis, syphilis, toxoplasmosis, rubella, hepatitis B infection, hepatitis C infection and human immunodeficiency virus infection among pregnant women attended at the Hospital Universitário Regional Norte do Paraná, Londrina State University, Paraná, a retrospective study of the serologic results performed in the prenatal routine during the period of June 1996 to June 1998 was carried out. The rates of seropositivity were as follows: american trypanosomiasis = 0.9%, syphilis = 1.6%, toxoplasmosis = 67% (IgG) and 1.8% (IgM), rubella = 89% (IgG) and 1.2% (IgM), hepatitis B surface antigen = 0.8%, hepatitis C virus = 0.8% and human immunodeficiency virus infection = 0.6%. An association between the increase in the seroprevalence of Chagas' disease and patient age was detected (p=0.006). The results underscore the importance of the serological tests in perinatal care, to prevent both the congenital and perinatally transmitted forms of theses infectious diseases.

Adolescent↗

Development of a real-time PCR assay to detect Treponema pallidum in clinical specimens and assessment of the assay's performance by comparison with serological testing.

The incidence of infectious syphilis in men who have sex with men and human immunodeficiency virus-infected patients has increased steadily in Victoria, Australia, since 2002. A TaqMan real-time PCR assay targeting the polA gene of Treponema pallidum (TpPCR) was developed. The analytical sensitivity of the assay was estimated to be 1.75 target copies per reaction. Initially, the assay was used to test a variety of specimens (excluding blood) from 598 patients. Of the 660 tests performed, positive PCR results were obtained for 55 patients. TpPCR results were compared with serology results for 301 patients being investigated for early syphilis. Of these patients, 41 were positive by both TpPCR and serology, 246 were negative by both TpPCR and serology, 4 were TpPCR positive but negative by serology, and 10 were TpPCR negative but showed evidence of recent or active infection by serology. Directly compared with serology, TpPCR showed 95% agreement, with a sensitivity of 80.39% and a specificity of 98.40%. Potential factors leading to the discrepant results are discussed. Concurrent serology on 21 patients with TpPCR-positive primary syphilitic lesions demonstrated that a panel of current syphilis serological tests has high sensitivity for the detection of early syphilis. We found that TpPCR is a useful addition to serology for the diagnosis of infectious syphilis. Direct comparison with other T. pallidum PCR assays will be required to fully assess the limitations of the assay.

Bacterial Proteins↗

Comparison of four serological tests for the detection of specific immunoglobulin M in cord sera of infants congenitally infected with cytomegalovirus.

Four serological tests (3 immunoassays using enzyme-labelled antigen and 1 radioimmunoassay) were compared as regards the detection of cytomegalovirus (CMV) immunoglobulin M in cord sera. 68 cord sera from infants congenitally infected by CMV were included in the study. The infections were primarily diagnosed by virus isolation close to birth. Four laboratories in 3 countries were involved, each laboratory using its own or a commercial test. Of the sera tested in the different laboratories 50-80% were found to be reactive. Both qualitatively and quantitatively there was a good correlation between the 3 enzyme-immunoassays. The RIA results differed to some extent from the enzyme tests as regards the quantification of IgM. The advantage of prospective IgM screening in undiluted cord sera followed by confirmatory virus isolation test in the neonatal period in IgM-positive cases is discussed.

Cytomegalovirus↗

Serodiagnosis of Helicobacter pylori infection: comparison and correlation between enzyme-linked immunosorbent assay and rapid serological test results.

CLOser is a new, one-step, qualitative anti-Helicobacter pylori immunoglobulin G test having the advantage of convenience and simplicity. We aimed to evaluate its diagnostic accuracy and to compare it with a quantitative enzyme-linked immunosorbent assay (ELISA) (HEL-pTEST II) in a study of 86 adult dyspeptic patients by using the results from histology and urease testing of gastric biopsies as a "gold standard." Forty-six patients were H. pylori positive. The sensitivities, specificities, and positive and negative predictive values were 95.7, 72.5, 80.0, and 93.5%, respectively, for CLOser and 93.5, 92.5, 93.5, and 92.5%, respectively, for HEL-pTEST II. The grade of the colored test bands in CLOser was correlated with antibody titers in HEL-pTEST II (r = 0.71; p < 0.001). The mean antibody titers were 13, 74, 186, and 328 U/ml for the negative, faint, thin, and thick bands, respectively, of CLOser. We concluded that the CLOser rapid serological test yielded sensitivity similar to that of the conventional ELISA. Although CLOser is not suitable for epidemiologic screening for H. pylori infection on account of lower specificity, it is particularly convenient and very easy to perform. Therefore, it may eventually become widely used in the office-based care of patients and lead to more cost-effective patient management decisions.

Adult↗