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Clinical utility of cytomegalovirus (CMV) serology testing in high-risk CMV D+/R- transplant recipients.

Late-onset cytomegalovirus (CMV) disease is a significant problem in D+/R- solid organ transplant (SOT) patients who receive antiviral prophylaxis. We assessed the clinical utility of CMV IgG and IgM serology testing for predicting late-onset CMV disease. We evaluated 352 D+/R- transplant recipients who participated in a trial comparing 100 days of ganciclovir versus valganciclovir prophylaxis. CMV serology was assessed on day 28, 56, 100, and 6 and 12 months post-transplant. IgG seroconversion occurred in 26.9% of patients by day 100, and in 63.4% and 75.3% by 6 and 12 months, respectively. IgM seroconversion occurred in 8.3%, 41.8% and 54.9% by day 100, month 6 and month 12, respectively. Seroconversion by day 100 (end of prophylaxis) was not predictive of subsequent CMV disease (CMV disease 13.3% if seropositive vs. 17.8% if seronegative; p = NS). However, at 6 months post-transplant, IgG serostatus was predictive of subsequent CMV disease between month 6 and 12 (CMV disease 1.3% if seropositive vs. 10.0% if seronegative; p = 0.002). In D+/R- patients, CMV serology testing is for the most part not clinically useful for predicting subsequent disease. However, seroconversion by 6 months may be useful for identifying patients at risk of late-onset CMV disease.

Antiviral Agents↗

Comparison of serologic tests for detection of antigen in canine heartworm infections.

In 30 random-source dogs, we determined sensitivity and specificity of 5 serologic tests for detection of canine heartworm antigens. Seventeen of the dogs were infected naturally with adult Dirofilaria immitis, and 4 of the infected dogs were amicrofilaremic. The ability of the serologic tests to predict whether a dog was infected or uninfected (overall test accuracy) ranged from 73 to 97%. Sensitivity was not affected by circulating D immitis microfilariae, but was markedly influenced by the number of adult D immitis present. False-positive reactions were rare and were not associated with intestinal parasites or Dipetalonema reconditum microfilariae. Modifications of some of the test procedures were necessary to maximize test accuracy and reproducibility. These modifications and other technical details might limit the usefulness of some of the tests in a veterinary practice.

Animals↗

Validity of using serological tests for diagnosis of diseases in wild animals.

Transposition of diagnostic tests used in domestic livestock species to free-ranging and captive wildlife species has two problems. First, most existing tests have not been adequately validated in domestic livestock. Second, assumptions that a serological test will perform identically in wildlife and livestock species may not be correct, due to differences in pathogenic strains and serovars, host serological responses, and exposure to organisms of similar antigenic structure which produce cross-reacting antibodies. Some assays require species-specific reagents/test components which might not be commercially available, and most assays have not been standardized. The authors outline the principles involved in the evaluation of a serological diagnostic test, and provide examples of how knowledge of test sensitivity and specificity can be used to estimate true prevalence, to determine whether a population is infected, and to facilitate management decisions with regard to animal translocations.

Animals↗

Chronic biological false-positive reactions to serological tests for syphilis in blood donors.

Problem sera from 375 blood donors were investigated for biological false-positive reactions to serological tests for syphilis. Problem sera are those sera in which submitting laboratories have found a reactive result to a screening test for syphilis. On each serum a cardiolipin Wassermann reaction, a Venereal Disease Research Laboratory test, a Reiter protein-complement fixation test, a Treponema pallidum immobilization test, and a fluorescent treponemal antibody absorption test were performed. Of the sera 49.9% were found to be non-reactive in all five tests, 28.8% gave results indicating a diagnosis of syphilis, and 21.3% showed biological false-positive reactions. There were 80 sera from blood donors which gave biological false-positive reactions. A further specimen of serum from 67 of these donors was tested after an interval of a minimum of six months. Sixty-six of the sera showed chronic biological false-positive reactions. Some donors were only found reactive for the first time after they had given very many donations of blood. It is concluded that sudden blood loss, as in blood donation, appears to stimulate the production of excess reagin in certain individuals, causing a chronic biological false positive reaction to serological tests for syphilis. This may only appear after several blood donations have been made.

Blood Donors↗

Routine serologic testing for hepatitis C virus infection should be instituted among dialysis patients.

Since the cloning of hepatitis C virus (HCV), a number of serologic and virologic tests for detecting infections with this virus have been made available for clinical practice. This led to the recognition of HCV as a major health hazard in hemodialysis (HD) centers. Yet the nephrology community has not been offered explicit recommendations about routine serologic testing for HCV of dialysis patients and consequently has remained unclear and divided about the utility of this testing. This review presents evidence in support of instituting routine serologic testing for HCV among dialysis patients. It concludes that because of the peculiar features of HCV infection in HD patients, serologic testing for HCV is the only means for unequivocal diagnosis of hepatitis C and an irreplaceable instrument for monitoring the incidence and the prevalence of the infection in this population. Serologic surveillance for HCV infection is crucial for identifying the association of new cases of HCV infection with potential risk factors for HCV transmission, particularly nosocomial, that can be modified in a way to prevent further dissemination of the virus. Data collected from routine serologic testing for HCV will undoubtedly improve our understanding of the epidemiology of HCV in the dialysis population and will provide a strong foundation for developing preventive measures and infection control strategies that are highly effective in controlling HCV infection. Routine serologic testing for HCV is also of direct benefit to the individual patient since establishing the HCV serologic status of each patient is important for optimizing patient care.

Alanine Transaminase↗

Efficacy of several serological tests and antigens for diagnosis of bovine brucellosis in the presence of false-positive serological results due to Yersinia enterocolitica O:9.

Yersinia enterocolitica O:9 bears a smooth lipopolysaccharide (S-LPS) of Brucella sp. O-chain A+C/Y epitopic structure and is a cause of false-positive serological reactions (FPSR) in standard tests for cattle brucellosis. Brucella S-LPS, cross-reacting S-LPSs representing several O-chain epitope combinations, Brucella core lipid A epitopes (rough LPS), Brucella abortus S-LPS-derived polysaccharide, native hapten polysaccharide, rough LPS group 3 outer membrane protein complexes, recombinant BP26, and cytosolic proteins were tested in enzyme-linked immunosorbent assays (ELISA) and precipitation tests to detect cattle brucellosis (sensitivity) and to differentiate it from FPSR (specificity). No single serological test and antigen combination showed 100% sensitivity and specificity simultaneously. Immunoprecipitation tests with native hapten polysaccharide, counterimmunoelectrophoresis with cytosolic proteins, and a chaotropic ELISA with Brucella S-LPS were 100% specific but less sensitive than the Rose Bengal test, complement fixation, and indirect ELISA with Brucella S-LPSs and native hapten or S-LPS-derived polysaccharides. A competitive ELISA with Brucella S-LPS and M84 C/Y-specific monoclonal antibody was not 100% specific and was less sensitive than other tests. ELISA with Brucella suis bv. 2 S-LPS (deficient in C epitopes), Escherichia hermannii S-LPSs [lacking the contiguous alpha-(1-2)-linked perosamine residues characteristic of Y. enterocolitica S-LPS], BP26 recombinant protein, and Brucella cytosolic fractions did not provide adequate sensitivity/specificity ratios. Although no serological test and antigen combination fully resolved the diagnosis of bovine brucellosis in the presence of FPSR, some are simple and practical alternatives to the brucellin skin test currently recommended for differential diagnosis.

Animals↗

HIV seroincidence estimates among at-risk populations in Buenos Aires and Montevideo: use of the serologic testing algorithm for recent HIV seroconversion.

Using the serological testing algorithm for recent HIV seroconversion, we estimated annualized incidences (per 100 person-years) of HIV-1 infection in different at-risk groups in Buenos Aires and Montevideo, during a 5-year period between 1998 and 2003. HIV-positive serum samples from 9 serosurveys conducted among men who have sex with men, patients attending clinics for a sexually transmitted infections consult (STIs), female commercial sex workers, injecting drug users (IDUs), noninjecting cocaine users (NICUs), asymptomatic women screened for HIV infection, and patients with tuberculosis were used. HIV incidences were as follows: 6.7 for men who have sex with men, 2.0 for STIs, 1.3 for female commercial sex workers, 0.0 for Argentinean IDUs, 10.3 for Uruguayan IDUs, 3.1 for Argentinean NICUs, 4.4 for Uruguayan NICUs, and 2.4 for patients with tuberculosis. Among asymptomatic women screened for HIV infection, incidence rose from 0.4 in 1998 to 4.6 in 1999 and to a high of 10.2 in the year 2000. Unexpectedly, high HIV incidences were detected among at-risk groups in Buenos Aires and Montevideo. This pattern shows an emerging HIV epidemic among heterosexuals stemming from core HIV-infected at-risk groups. There is an urgent need for development and implementation of specific prevention strategies to address this burgeoning epidemic.

Algorithms↗

Serum sperm antibodies in cases of azoospermia: comparative diagnostic value of separate and combined, agglutination, immoblization and cytotoxic serological tests.

To study the diagnostic value of various serological tests in the detection of serum sperm antibodies a group of 50 patients with azoospermia was studied. Cases were divided by testicular biopsy into: 32 obstructive and 16 non obstructive cases. Tests applied included a microagglutination and a macroagglutination test as well as 2 complement dependent tests; a sperm cytotoxic and a sperm immobilization tests. Applied separately the macroagglutination was the most sensitive test but used together, more cases were detected. Because of this and the different value and significance of each test, it is advisable to use more than one test in the investigations of cases of male infertility.

Adult↗

Staining toxoplasma gondii with fluorescein-labelled antibody. II. A new serologic test for antibodies to Toxoplasma based upon inhibition of specific staining.

A new serologic test for antibodies to Toxoplasma is described, which is based upon inhibition of specific staining with fluorescent antibody. In performing the test, a mixture of the test serum and known fluorescein-labelled antiserum is added to a dried smear of toxoplasms for 1 hour at 37 degrees C. The smear is then rinsed and examined with a fluorescence microscope. Reduction in the brightness of fluorescence, as compared to that of a negative control slide, indicates the presence of antibody in the test serum. A comparison of the results of this test with those of the methylene blue dye test showed a strong parallelism between the two sets of results. On the other hand, the complement-fixation test for toxoplasmosis did not yield nearly as many positives as the inhibition test. The specificity of the new test was studied by comparing it with dye test results and clinical histories in human patients, and by testing a group of animals immunized with a variety of non-Toxoplasma antigens. No evidence of cross-reactions was obtained in the latter series. Some advantages and disadvantages of the inhibition test are discussed.

Animals↗

Contribution of serological tests and blood culture to the early diagnosis of systemic candidiasis.

The isolation of Candida species from a single blood culture is considered sufficient evidence for the initiation of systemic antifungal therapy. However, blood cultures still lack sensitivity. Previous reports have suggested that the combined serological detection of mannanemia and anti-mannan antibodies may be useful for the diagnosis of systemic candidiasis caused by Candida albicans (specificity and sensitivity 93% and 80%, respectively). In this study, serological tests to detect Candida albicans mannan and Candida albicans antibodies (Platelia Candida Antigen and Antibody tests; Bio-Rad, France) were applied retrospectively to a series of patients with at least one Candida-positive blood culture and from whom at least one serum sample, taken before or on the day of blood culture, was available. Forty-five patients were selected, including 23 infected by Candida albicans, 4 by Candida glabrata, 9 by Candida tropicalis, 5 by Candida parapsilosis, and 4 by Candida krusei. Serological tests were positive in 73% of patients at least 2 days, and in some patients, up to 15 days before blood cultures became positive. These data suggest that serological surveillance of at-risk patients using the Platelia Candida tests could result in earlier initiation of antifungal therapy, especially when used in conjunction with blood cultures. In this way, more efficient management of nosocomial infections caused by Candida species can be achieved.

Candidiasis↗

One-off streptococcal serologic testing in young children with recurrent tonsillitis.

OBJECTIVES: Recurrent acute tonsillitis in children under 4 years of age is usually viral, making antibiotic therapy inappropriate and the indication for tonsillectomy uncertain. Identifying those young children with bacterial infections is therefore important. The purpose of this study was to determine whether one-off streptococcal serologic testing is a useful tool in assessing recurrent acute tonsillitis in young children. METHODS: We performed a retrospective study of 45 children (35 male and 10 female) under the age of 4 years who were found by a staff otolaryngologist to have recurrent acute tonsillitis over a 5-year period and had one-off serologic testing for anti-streptolysin O titers and anti-deoxyribonuclease B levels. Data were collected by chart review. RESULTS: Three children (6.7%) had clearly positive titers for either one or both streptococcal antibodies. Children with negative serologic results were significantly less likely to have shown a significant response to antibiotic therapy for their acute episodes (26% versus 100%; p = .026). Nine children (20%) eventually underwent tonsillectomy, all of whom had negative serologic results. CONCLUSIONS: Anti-streptolysin O and anti-deoxyribonuclease B levels may aid clinical evaluation of recurrent acute tonsillitis in young children in differentiating between those cases due to group A beta-hemolytic Streptococcus and those that are viral in origin.

Acute Disease↗

Serologic testing for Trypanosoma cruzi: comparison of radioimmunoprecipitation assay with commercially available indirect immunofluorescence assay, indirect hemagglutination assay, and enzyme-linked immunosorbent assay kits.

The radioimmunoprecipitation assay (RIPA) has been used as a confirmatory test in several ongoing and published studies of Trypanosoma cruzi in blood donors in the United States. Despite its use as a confirmatory test, few studies are available comparing RIPA to commercially available serologic test methods. Thus, we compared RIPA with two indirect hemagglutination assays (Biolab Diagnostica SA, São Paulo, Brazil; Hemagen Diagnostics, Inc., Waltham, Mass.) and four different enzyme-linked immunosorbent assays (Abbott Laboratories, Abbott Park, Ill.; Embrabio, São Paulo, Brazil; Organon Teknika, São Paulo, Brazil; and Gull Laboratories, Salt Lake City, Utah) using a panel of 220 serum specimens from Brazilian blood donors with a range of T. cruzi antibody titers as determined by indirect immunofluorescence assay (IFA). A titer of 1:20 was used as the baseline for seropositivity. All IFA-negative serum specimens (n = 19) were nonreactive on all tests. At a titer of 1:20 (n = 9), reactivity rates varied considerably among the tests, with only the RIPA and the Organon and Gull assays identifying reactive specimens. For specimens at a 1:40 titer (n = 35), most assays identified at least 32 of 35 (91%) specimens as reactive, but the Biolab assay only identified 24 (69%). At higher titers (1:80, n = 56; 1:160, n = 101) the assays were comparable, with the exception of the Biolab assay, demonstrating rates of agreement with IFA of >/=98%. Overall, when compared with several other test formats, RIPA demonstrated equivalent or superior rates of agreement with IFA-positive specimens across all titers examined. In particular, at titers of >1:40, the RIPA compared favorably with other test methods currently in use, supporting its application as a confirmatory test, particularly in a research setting.

Animals↗

Simple serological tests for detecting classical heat labile enterotoxin (LT-I) of Escherichia coli.

Diarrhoea caused by enterotoxigenic Escherichia coli (ETEC) remains a problem in Southeast Asia. At present, no routine laboratories as yet are available for ETEC detection. In this study, attempts were made to produce reagents for use in simple serological tests for detecting LT. The serological methods were the Biken, the staphylococcal coagglutination and the reverse passive hemagglutination tests. For the Biken test, medium was prepared locally by mixing constituents as described previously by Honda et al., (1981). Anti-CT-B subunit was prepared by immunizing a rabbit with commercial CT-B subunits (Sigma). Other chemical reagents e.g. colistin, lincomycin etc. were obtained from the local supplies. Using the locally made reagents to detect LT from 100 WHO reference strains of E. coli by the Biken test, it was found that the test had 100%, 92%, 96%, 100% and 92.5% of specificity, sensitivity, accuracy, positive predictive value and negative predictive value, respectively. Protein A rich Staphylococcus aureus from the stock culture of the Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University were grown in suitable medium i.e. blood agar containing lincomycin (BA-Lin). Suitable amount of the rabbit anti CT-B subunit (0.1 ml) was used to sensitize each ml of the formalinized, heat-fixed bacteria. The sensitized bacteria were used for detecting LT in the lysates of the 100 E. coli reference strains. The lysates were prepared by growing the E. coli strains on BA-Lin medium for 8 hours, then a loopful of each strain was inoculated into colistin solution (20,000 unit/ml).(ABSTRACT TRUNCATED AT 250 WORDS)

Agglutination Tests↗

[Comparative study on three serologic tests for detecting antibodies in cases with bancroftian microfilaremia].

A comparative study was conducted for detecting antibody levels of bancroftian microfilaremia cases by using three serologic tests, IFAT, ELISA and dot-ELISA. A total of 102 serum specimens were collected from bancroftian microfilaremia cases. The positive rate was 89.2% (91/102) with IFAT (serum dilution > or = 1:20), 84.3% (86/102) with ELISA (OD value > or = 0.440) and 89.2% (91/102) with dot-ELISA (serum dilution > or = 1:80), respectively (P > 0.05); while in healthy individuals from non-endemic areas the false positive rate was 0(0/30), 1.8(1/55) and 0(0/54), respectively. The results indicated that there were no statistically significant differences among the three serologic tests. There was no correlation found in this study between microfilaria density and the antibody level. It was also proved that the detection rate of the combined use of either 2 or 3 tests was higher than each of the three tests; the positive rates being 94.1%-98.0%.

Animals↗

Usefulness of a new serological test (Bio-Rad) to diagnose Helicobacter pylori-associated gastritis.

A semiquantitative serological test (G.A.P. test; Bio-Rad) to diagnose Helicobacter (Campylobacter) pylori-associated gastritis has recently become commercially available. This test was evaluated with sera from 72 dyspeptic patients with known H. pylori status. When the instructions provided by the manufacturer were applied strictly, the sensitivity of the test was only 32.7%. With a slight modification of the instructions, the performance characteristics were still unsatisfactory (specificity, 43.5%). When a new cutoff value was determined by a receiver operating curve, the quality of the test (positive predictive value, 95.6%; negative predictive value, 77.8%) was slightly lower than that of a noncommercial enzyme-linked immunosorbent assay (positive predictive value, 95.7%; negative predictive value, 84.0%). It is concluded that the Bio-Rad G.A.P. test has low diagnostic yield, and it is stressed that commercial tests should be evaluated extensively before being applied in daily practice.

Antibodies, Bacterial↗

[Comparative evaluation of the commercial Sevatest ELISA immunoenzyme set and other serological tests for detecting Toxoplasma gondii antibodies].

This work analyzes the results of 4 serologic tests used for the diagnosis of toxoplasmosis: the complement fixation (CFT), indirect immunofluorescence (IIF), passive hemagglutination (PHAT) tests, and the enzyme-linked immunosorbent assay (ELISA). The last-mentioned one was made with the use of the commercial kits Sevatest ELISA IgG/Toxo Micro I. The results of ELISA were in good correlation with those yielded by the traditional tests: 70% coincidence with CFT, 80% with IIF, 84% with PHAT; besides, ELISA has shown a higher sensitivity in the screening of sera.

Antibodies↗

Specificity of routine parasite serological tests in autoimmune disorders, neoplastic disease, EBV-induced mononucleosis, and HIV infection.

Sera from 120 patients with rheumatological disorders, neoplastic disease, infectious mononucleosis, and HIV infection, and from 30 healthy blood donors were tested for nonspecific reactivity in 13 routinely used parasite serological tests. Responses were detected in 3/30 healthy blood donors (10%) vs 25/120 patients (21%). Of 40 responses in these 28 responders most were weakly reactive, and 25 out of 40 responses were only at borderline level. Response rates were highest in patients with mononucleosis presumably due to heterophile antibodies. Only four patients had responses in at least two different serodiagnostic tests for the same parasitic infection. Response patterns indicative of a possible underlying, concurring, or superimposed parasitic infection, thus, were rare. Especially susceptible to nonspecific reactivity seemed to be immunofluorescent antibody tests for filariasis, schistosomiasis, and amebiasis. In conclusion, compared to healthy controls, false-positive serological responses seem to be more frequent in certain disease groups dependent on the test methods used. Second, the use of more than one serodiagnostic test for the same parasitic disease will substantially facilitate the identification of nonspecific reactivity. Third, for better defining the specificity of parasitological serodiagnosis, more studies should include control sera from patients with nonparasitic diseases that frequently show immunological abnormalities.

Acquired Immunodeficiency Syndrome↗

Serological testing--an alternative to boosters?

The issue of the duration of immunity, particularly for the modified live viral components of veterinary vaccines, has been a significant part of the recent vaccination debate. One manufacturer has increased the recommended booster interval for these components to 3 years give name and another now states 'up to 4 years' immunity. There remain many unanswered questions regarding this duration of immunity (DOI). Studies suitable for data sheet claims are time consuming and costly and can only be performed in laboratory dogs under tightly controlled conditions. Evidence from rabies serology testing in the UK shows that the response of individual animals to routine vaccination is highly variable. Much of the published field evidence on the persistence of antibody titres originates from North America, where vaccination strategies and reservoir species differ from Europe. Quantifying the effect of exposure to field virus on the maintenance of immunity in these studies is impossible, and little is known of the circulation of virus in unvaccinated dogs and wild mammals throughout Europe. If owners or vets are concerned about revaccination one option is to assess the need for each booster by performing a blood test. There is some published evidence of the relationship between antibody titres and protective immunity, and tests are available to measure responses to individual viral components of the routine canine and feline vaccines. It must be remembered that most commercial tests to assess immunity only measure antibodies, which are only one aspect of the immune response to vaccination. It is therefore possible that animals without or with low antibody titres are in fact protected. Serological tests are an option if owners are unwilling to have their animal boostered without evidence that it is needed. However, the cost of these tests is likely to exceed that of booster vaccination for the foreseeable future.

Animals↗