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Evaluation of four commercially available rapid serologic tests for diagnosis of leptospirosis.

Four rapid tests for the serologic diagnosis of leptospirosis were evaluated, and the performance of each was compared with that of the current standard, the microscopic agglutination test (MAT). The four rapid tests were a microplate immunoglobulin M (IgM)-enzyme-linked immunosorbent assay (ELISA), an indirect hemagglutination assay (IHA), an IgM dipstick assay (LDS), and an IgM dot-ELISA dipstick test (DST). A panel of 276 sera from 133 cases of leptospirosis from four different geographic locations was tested as well as 642 sera from normal individuals or individuals with other infectious or autoimmune diseases. Acute-phase sera from cases (n = 148) were collected or=15 days after onset (median = 29.1). By a traditional method (two-by-two contingency table), the sensitivities for detection of leptospirosis cases were 93.2% by LDS, 92.5% by DST, 86.5% by ELISA, and 79.0% by IHA. Specificity was 98.8% by DST, 97% by ELISA and MAT, 95.8% by IHA, and 89.6% by LDS. With a latent class analysis (LCA) model that included all the rapid tests and the clinical case definition, sensitivity was 95.5% by DST, 94.5% by LDS, 89.9% by ELISA, and 81.1% by IHA. The sensitivity and specificity estimated by the traditional methods were quite close to the LCA estimates. However, LCA allowed estimation of the sensitivity of the MAT (98.2%), which traditional methods do not allow. For acute-phase sera, sensitivity was 52.7% by LDS, 50.0% by DST, 48.7% by MAT and ELISA, and 38.5% by IHA. The sensitivity for convalescent-phase sera was 93.8% by MAT, 84.4% by DST, 83.6% by LDS, 75.0% by ELISA, and 67.2% by IHA. A good overall correlation with the MAT was obtained for each of the assays, with the highest concordance being with the DST (kappa value, 0.85; 95% confidence interval [CI], 0.8 to 0.90). The best correlation was between ELISA and DST (kappa value, 0.86; 95% CI, 0.81 to 0.91). False-positive LDS results were frequent (>or=20%) in sera from individuals with Epstein-Barr virus, human immunodeficiency virus, and periodontal disease and from healthy volunteers. The ease of use and significantly high sensitivity and specificity of DST and ELISA make these good choices for diagnostic testing.

Humans↗

Guidelines for serological testing in the diagnosis of acute hepatitis A and B.

This study was performed to determine whether elevation of serum transaminases can be used to eliminate unnecessary serological tests to diagnose acute hepatitis A (HAV) and acute hepatitis B (HBV). Serum samples of 1226 patients were tested for HBsAg, anti-HBc (IgM), and anti-HAV (IgM). Acute hepatitis was diagnosed in 113 (9.2%) patients; 75 were serologically positive for HAV, 36 for HBV, and 2 patients for both HAV and HBV. Serum transaminase levels were elevated in 104 of 107 (97.2%) of seropositive patients in whom the results of biochemical tests were available. A review of the medical records of seropositive patients with normal transaminases revealed that each of the three HAV patients had a remote history of hepatitis. None of the seropositive patients with a recent history of acute viral hepatitis had normal transaminase levels. During this period, serological tests were ordered in 266 of 1054 (25.2%) seronegative patients with normal serum transaminases. We conclude that serum transaminase levels can be reliably used to screen sera for acute HAV and HBV infection.

Acute Disease↗

Evaluation of serological tests for screening of chlamydial eye diseases.

Trachoma is recognized as one of most important origins of blindness in developing countries and inclusion conjunctivitis is associated with STD in developed countries. We evaluated the diagnostic value of serological tests for the screening of eye diseases associated with Chlamydia trachomatis infection. We determined serum IgG, IgA and IgM antibodies to C. trachomatis from 53 Japanese patients with active inflammatory trachoma (aged more than 60 years) and from 107 adult patients (aged 20 to 50 years) with acute inclusion conjunctivitis by ELISA test kit. We detected serum IgG antibodies from 22 out of 53 (42.5%) patients with trachoma and from 40 out of 107 (37.4%) patients with acute inclusion conjunctivitis. We also detected serum IgM antibodies from 7 out of 53 (13.2%) patients with trachoma and from 35 out of 107 (32.7%) patients with acute inclusion conjunctivitis. The prevalence of serum IgM antibodies to C. trachomatis in patients with acute inclusion conjunctivitis was significantly higher than that in patients with active trachoma (p < 0.05). Serological tests are also thought to be useful for screening of chlamydial eye diseases.

Adult↗

Use of locally isolated saprophytic Leptospira strain for serological testing of human leptospirosis.

A saprophytic Leptospira isolate recovered from tap water was utilized for serological testing. One hundred-twenty Serum samples comprising 55 cases from PUO/febrile jaundice and 65 samples from apparently healthy individuals were tested by MAT and HA using this environmental saprophytic strain and the results compared with that of Leptospira biflexa semaranga patoc, the standard saprophytic strain commonly employed for sero-diagnosis of leptospirosis. The MAT data showed 96.4 per cent correlation between the two strains. Similarly, the HA results were matching to the extent of 94.5 per cent. Results, therefore, suggest that local saprophytic Leptospira strain may serve as a substitute to serovar patoc for serodiagnosis of leptospirosis.

Antibodies, Bacterial↗

Immunologic and serologic testing in patients with Ménière's disease.

OBJECTIVE: To establish the value of immunologic and serologic testing in patients with Ménière's disease. STUDY DESIGN: Prospective cohort. SETTING: University-based, tertiary care balance center. INTERVENTION: Patients with active unilateral or bilateral Ménière's disease underwent testing, including antinuclear antibodies, anti-DNA antibodies, rheumatoid factor, complement levels, anti-Sjögren syndrome A and B antibodies, sedimentation rate, antiphospholipid antibodies, Western blot for anticochlear antibodies (anti-heat shock protein 70), microhemagglutination test for Treponema pallidum, and Lyme titers. OUTCOME MEASURES: Results of laboratory tests. RESULTS: In patients with unilateral Ménière's disease (n = 40), 27% demonstrated elevated antiphospholipid antibody titers (population norm, 6-9%). The majority of these patients manifested negative assays on the other tests listed above. In patients with bilateral Ménière's disease (n = 18), elevations in antinuclear antibody titers (38%) were the most notable finding. As part of a broader study of patients with progressive hearing loss, four patients with positive syphilis titers were identified; however, none of these patients complained of vertigo. CONCLUSION: In general, the results of this study do not support the hypothesis that immune or infectious pathologies are involved in the pathogenesis of unilateral Ménière's disease. In particular, Lyme disease does not seem to cause labyrinthine disease. However, the potential role of the thrombogenic antiphospholipid antibodies must be further investigated. Patients with bilateral Ménière's disease may be more likely to have a systemic autoimmune process.

Antibodies, Antinuclear↗

Polymerase chain reaction in comparison with serological tests for early diagnosis of human leptospirosis.

The aim of this study was to compare the sensitivity and specificity of polymerase chain reaction (PCR) using two primer pairs and combined with blood culture, immunoglobulin M enzyme-linked immunosorbent assay (IgM ELISA), microscopic agglutination test (MAT) and slide agglutination test (SAT) in the diagnosis of human leptospirosis. We analysed 124 serum samples: 60 from patients with confirmed leptospirosis, 20 from patients with other diseases and 44 from healthy individuals. Analysing the first serum sample collected during the first 3-8 days of disease, the sensitivities of the four tests MAT, IgM ELISA, SAT and PCR were, respectively, 69.0%, 79.3%, 72.4% and 62%. In subsequent samples, those same sensitivities were, respectively, 95.4%, 100%, 100% and 72.7% in samples collected from days 9 to 14 and 88.9%, 88.9%, 77.8% and 44.4% in those collected from days 15 to 42. The most specific method (at 100%) was PCR and the least specific (at 89.1%) was IgM ELISA. Although we found PCR to be less sensitive than the serological tests over the course of the disease, our data indicate that PCR was the most sensitive in those initial serum samples presenting no specific antibodies detectable by any of the serological methods tested. We also recommend that PCR can be used in combination with serological tests as we found that this improves the sensitivity of the diagnosis of leptospirosis in the first phase of the disease (93.1-96.5%).

Adult↗

Limitation of serological testing for Lyme borreliosis: evaluation of ELISA and western blot in comparison with PCR and culture methods.

The aim of the study was to evaluate a one-step procedure using an ELISA test of high specificity and a two-step procedure using immunoblot as a confirmation test, and to compare the results of serological testing with detection of bacterial DNA and living spirochetes. Sera, synovial (SF) and cerebro-spinal fluids (CSF) were obtained from 90 patients with clinical symptoms of Lyme borreliosis. Serum samples were tested with recombinant ELISA and Western blot assay. Citrated blood, cerebrospinal and synovial fluids samples were cultured in cell line and tested by PCR to detect spirochetes. No correlation was found between levels of specific B. burgdorferi antibodies detected with a recombinant antigen ELISA and the number of protein fractions developed with these antibodies by immunoblot. Moreover, Lyme borreliosis patients who have live spirochetes in body fluids have low or negative levels of borrelial antibodies in their sera. This indicates that an efficient diagnosis of Lyme borreliosis has to be based on a combination of various techniques such as serology, PCR and culture, not solely on serology.

Bacteriological Techniques↗

Use of the serologic testing algorithm for recent HIV seroconversion (STARHS) to identify recently acquired HIV infections in men with early syphilis in Los Angeles County.

BACKGROUND: Syphilis outbreaks among men who have sex with men (MSM) in the United States, many of whom are HIV infected, have prompted increased concern for HIV transmission. METHODS: To identify whether men are acquiring HIV concomitantly or within the critical period of syphilis infection, banked Treponema pallidum particle agglutination-positive serum specimens from men with early syphilis infection were screened for HIV-1 antibody. Samples that were positive for HIV antibody were then tested with a less sensitive (LS) HIV-1 antibody enzyme immunoassay (serologic testing algorithm for recent HIV seroconversion [STARHS]) to identify HIV infections that occurred on average within the previous 6 months. RESULTS: Of the 212 specimens banked from men with early syphilis, 74 (35%) were HIV-positive. Of these, 15 tested non-reactive by the LS assay. Twelve of these 15 were considered to be recent infections by the LS assay and testing history. Eleven (92%) of the recent infections were among MSM. One man had primary syphilis, 6 (50%) had secondary syphilis, and 5 (42%) had early latent syphilis. Eight men (67%) reported sex with anonymous partners, and 3 (25%) reported consistent condom use. The estimated HIV incidence was 17% per year (95% confidence interval [CI]: 12%-22%) among all men with early syphilis, and it was 26% per year (95% CI: 91%-33%) among MSM. CONCLUSIONS: Syphilis epidemics in MSM may be contributing to HIV incidence in this population. The STARHS can be applied as a surveillance tool to assess HIV incidence in various at-risk populations, but further studies are necessary for validation.

Adult↗

Use of viral cultures and serologic tests for cytomegalovirus infection. Rational or random?

To assess their applied clinical utility, viral cultures and serological tests for cytomegalovirus (CMV) were reviewed at Duke University Medical Center (DUMC), a 1,125-bed tertiary-care hospital. Less than 1% (3 of 1,216) of CMV cultures were positive, and 8% of serum samples (45 of 587) were positive by single sera IgM ELISA. Sixteen percent (32 of 199) of IgG acute to convalescent sera pairs were positive. Four hundred five of 588 (69%) serum samples were positive for the IgM/IgG passive latex agglutination test, consistent with the results for random blood donors. Review of hospital records showed that fewer than 1% of the positive-test patients (excluding the latex test) received treatment for CMV. Comparisons of tests ordered on individual patients did not disclose a coherent diagnostic strategy. The authors conclude that the majority of testing for CMV in their medical center does not yield useful clinical information, but carries a substantial financial burden. A new diagnostic strategy to attempt to diagnose CMV disease is needed.

Acute Disease↗

The value of serological tests for syphilis in atypical genital ulcer disease.

A 45-year-old heterosexual man presented with phimosis three weeks after arriving in this country from India. Examination under anaesthesia and circumcision revealed a large ulcer on the ventral aspect of the penis around the fraenum. Histopathological examination of a biopsy taken from the ulcer revealed granulation tissue with inflammatory cell infiltrate. Specific and non-specific serological tests for syphilis carried out four weeks after the initial presentation showed high titres. The importance of serological tests for syphilis in genital ulcer disease, especially in atypical presentations, is discussed.

Humans↗

Serologic testing in inflammatory bowel disease: its value in indeterminate colitis.

Serologic testing in inflammatory bowel disease provides a unique tool to classify these diseases into more homogeneous groups. Serum markers can also be used to identify common antigenic triggers and specific defects in mucosal immune regulation and cytokine imbalance in different parts of the gastrointestinal tract. ANCA (antineutrophil cytoplasmic antibodies) and ASCA (anti-Saccharomyces cerevisiae antibodies), the most extensively studied serologic markers, have been used to classify ulcerative colitis and Crohn's disease into such groups with certain phenotypic characteristics and responses to treatment. Similar studies have been initiated in indeterminate colitis, and these results will help in the future to define the immunopathogenesis, prognosis, and response to certain treatments in patients with this type of disease.

Animals↗

Diagnostic value of serological tests against Pseudomonas aeruginosa in a large cystic fibrosis population.

BACKGROUND: Serological methods to monitor Pseudomonas aeruginosa colonisation in patients with cystic fibrosis (CF) are advocated but the diagnostic value of a commercially available P aeruginosa antibody test to detect early and chronic P aeruginosa colonisation in a non-research setting has not been assessed. METHODS: Colonisation with P aeruginosa was estimated by regular culture of sputum or oropharyngeal swabs during three consecutive years in 220 patients with CF aged 0-65 years. Commercially available ELISA tests with three P aeruginosa antigens (elastase, exotoxin A, alkaline protease) were performed at the end of the study period. In a subgroup of 57 patients (aged 4-14 years) serological tests were performed annually. RESULTS: Using culture as the reference standard, the ELISA tests using the advised cut off values had a sensitivity of 79% and a specificity of 89% for chronic colonisation. Receiver-operator characteristic curves were created to optimise cut off values. Applying these new cut off values resulted in a sensitivity of 96% and a specificity of 79%. All three individual serological tests discriminated well between the absence and presence of chronic P aeruginosa colonisation. The sensitivity of the individual antibody test was 87% for elastase, 79% for exotoxin A, and 76% for alkaline protease. First colonisation was preceded by positive serological results in only five of 13 patients (38%). CONCLUSION: In patients with CF, serological tests using specific antigens are sensitive for diagnosing chronic P aeruginosa colonisation. However, the failure of serological tests to detect early colonisation in young patients emphasises the need for continued reliance on cultures.

Adult↗

Results of serological tests for syphilis among Gurkhas and other high risk groups.

Serological tests for syphilis gave more positive results in serving Gurkha (Nepali) soldiers from west Nepal than in those from east Nepal or in Gurkha recruits. The soldiers had served from four to 11 years. The source of their infection was not clear. Positive results were rather less common in black patients born in the tropics attending a genitourinary medicine in London and were similar to findings in blood donors in the West Indies. British born male patients attending a genitourinary medicine department in London had a much lower prevalence. Malay and Nepali women attending an antenatal clinic in Singapore had a higher prevalence of positive serological results than women attending an antenatal clinic in London. Nepalis, Malays, and black people born in the tropics continue to require serological screening.

Adolescent↗

Accuracy of supplementary serologic testing for human T-lymphotropic virus types I and II in US blood donors. Retrovirus Epidemiology Donor Study.

Blood donations in the United States have been screened for antibody to human T-lymphotropic virus type I (HTLV-I) by HTLV-I enzyme immunoassay (EIA) since November 1988. Specimens repeatedly found to be reactive by EIA undergo confirmation by supplementary serologic tests. We assessed the accuracy of blood center testing of 994 HTLV-I EIA repeat-reactive specimens in five US blood centers between November 1988 and December 1991. Of 410 confirmed HTLV-I/II donations, 407 (99.3%) were infected with HTLV-I/II, as determined by polymerase chain reaction (PCR) (403 cases) and by repeat serologic testing (4 cases). The three false-positive results occurred in the first year of testing. Of 425 HTLV-indeterminate specimens, 6 (1.4%) were found to be infected by PCR (5 with HTLV-II and 1 with HTLV-I). None of 159 confirmatory test-negative donations was PCR positive. Of HTLV-I/II-seropositive specimens, 80.2% to 95.4% could be typed as HTLV-I or HTLV-II by type-specific serologic assays. These results support recommendations that HTLV-I/II-seropositive donors should be advised that they are infected with HTLV-I, HTLV-II, or HTLV-I/II (depending on results of type-specific assays). HTLV-indeterminate donors should be advised that their results only rarely indicate HTLV infection. HTLV confirmatory test-negative donors should be reassured that they are not infected with HTLV-I or HTLV-II.

Blood Donors↗

Evaluation of HIV serial and parallel serologic testing algorithms in Abidjan, Côte d'Ivoire.

OBJECTIVE: To evaluate HIV serologic testing algorithms based on a combination of three enzyme linked immunosorbent assays (ELISA) for the confirmation of HIV infection in Abidjan, Côte d'Ivoire, where HIV-2 and HIV-1 non-B subtypes are prevalent. METHODS: A total of 1069 human sera with known serologic status, in addition to a seroconversion and low titer antibody panel were initially tested by six ELISA to determine the sensitivity, specificity and delta values of the assays. On the basis of the performance of the assays, three ELISA (Enzygnost, ICE 1.0.2, and Vironostika) were selected for use in a parallel and serial testing algorithm in analyzing 8283 consecutively collected sera. In the parallel testing algorithm, sera concordantly reactive or non-reactive by Enzygnost and ICE 1.0.2 were considered as true positive or true negative, respectively. In the serial algorithm, sera reactive by Enzygnost were retested by ICE 1.0.2. Sera with discordant results were tested by Vironostika, and the results was considered definitive. All reactive sera, plus a random sample of negative sera were tested for confirmation by Peptilav. In addition, a random sample of reactive sera was tested by Western blot. RESULTS: All ELISA had 100% sensitivity; specificities ranged from 96.8 to 100%. Positive and negative delta values of the ELISA were high (range, 6.89 to 46.07 and -2.05 to -5.75, respectively). Of the 8283 sera, 2054 were considered true positives and were correctly classified by the parallel testing algorithm (sensitivity, 100%). Of the 6229 true negative sera, 6226 were negative by the parallel testing algorithm (specificity, 99.95%). The sensitivity of the serial algorithm was 99.96%, and specificity was 99.95%. None of the 250 concordant ELISA-negative sera in the algorithm that were randomly tested in Peptilav was positive; similarly, all of the 103 concordant ELISA-positive sera were confirmed by Western blot. The three-ELISA algorithm resulted in reagent cost-savings of at least 50% compared with the Peptilav-based algorithm. CONCLUSION: These results suggest that a combination of ELISA using different principles or antigens in a serial or parallel algorithm is an efficient and cost-effective alternative to the standard algorithm in areas where HIV-1 and HIV-2 are prevalent.

Algorithms↗

[The evolution of serologic tests in the diagnosis of HCV infections].

The Authors take in account the technical aspects of the serological tests for the diagnosis of HCV infections. Particularly they consider the better diagnostic capacity of the 2nd generation and, at the present time, 3rd generation screening tests compared to tests of 1st generation. This happens because an antigen, the c33-c, codified by a genomic sequence, specific for the non-structural proteins, NS3, of the genome of HCV virus is used in the last methodologies. This antigen induces the production of specific antibodies more quickly than the other antigenic proteins. Moreover in many cases of HCV infections it has been evidenced that the anti-c33-c are the only antibodies produced. For this reason the tests that use this antigen not only allow an early diagnosis, enclosing the "window effect", but in every case have the best diagnostic capacity. Moreover the authors evaluate also the tests defined "confirmatory tests", in which the importance of the presence of systems for the identification of both anti-c33-c and c22-3 antibodies is evidenced.

Antigens, Viral↗

Serological tests in cerebrospinal fluid for congenital syphilis in central Africa.

Serological tests were performed in the cerebrospinal fluid (CSF) of 13 children with active congenital syphilis (presence of specific IgM FTA-ABS antibodies) and of seven seropositive children with no active syphilis (FTA-ABS IgM-negative) born to syphilitic treated mothers in Libreville, Gabon. Antibodies against treponema were measured by the Venereal Disease Research Laboratory test (VDRL), the Treponema pallidum haemagglutination assay (TPHA) and the fluorescent treponema antibody absorption tests (FTA-ABS IgG and IgM). Of the 13 children with active syphilis, seven had a positive FTA-ABS IgG in the CSF. The result of this test was not correlated with the severity of clinical features, CSF protein levels or number of CSF white blood cells. The CSF-TPHA test was positive in four out of 12 children, and the CSF-VDRL test was negative in all the children with active congenital syphilis. One of the seven newborns with mother-transmitted antibodies had a positive FTA-ABS and TPHA in the CSF. These data show that the VDRL is not sensitive enough to diagnose congenital neurosyphilis, and that FTA-ABS or, at least, TPHA are convenient, sometimes with false-positive results, when a sophisticated method of detecting specific IgM in CSF is not available.

Fluorescent Treponemal Antibody-Absorption Test↗