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A WWW-accessible knowledge base for the interpretation of hepatitis serologic tests.

HEPAXPERT is a knowledge-based system that interprets the results of routine serologic tests for infection with hepatitis A and B viruses. The following tests are included: hepatitis A virus anti-bodies (anti-HAV), IgM antibodies to the hepatitis A virus (IgM anti-HAV), hepatitis A virus (HAV in stool, hepatitis B surface antigen (HBsAg) and antibodies (qualitative anti-HBs, quantitative anti-HBs titre), antibodies to hepatitis B core antigen (anti-HBc and IgM anti-HBc), and hepatitis B envelope antigen (HBeAg) and antibodies (anti-HBe). HEPAXPERT/WWW--an implementation of HEPAXPERT-III for WWW--can be reached by URL http://www.swun.com/hepax of the World Wide Web. After selecting HEPAXPERT/WWW, serologic test results can be entered and will be transferred as an e-mail message for subsequent interpretation which is done off-line with HEPAXPERT-III. The textual interpretation is sent back via e-mail. Each qualitative test for hepatitis A and B antibodies and antigens may produce one of four possible results: positive, negative, borderline, and not tested. To cover the resulting 64 (A) and 57344 (B) combinations of findings, the knowledge base of HEPAXPERT/WWW contains 16 rules for hepatitis A and 131 rules for hepatitis B serology interpretation. This basic knowledge is structured such that all possible combinations of findings can be interpreted and there is no overlap in the premises underlying the rules. The reports that the system automatically generates include: (a) the transferred results of the tests; (b) a detailed analysis of the results, including virus exposure, immunity, stage of illness, prognosis, infectiousness, and vaccination recommendation; and (c) optional: an ID to distinguish the origin of the interpretation requests.

Artificial Intelligence↗

Comparison of three HCV genotyping assays: a serological method as a reliable and inexpensive alternative to PCR based assays.

BACKGROUND: Determination of hepatitis C virus (HCV) genotypes and subtypes is of rising clinical importance. In times where also an increasing need for cost effectiveness can be observed, the demand for fast and easy performable assays grows. OBJECTIVES: To evaluate and compare different genotyping methods regarding their reliability, practicability, and expense in the daily routine. METHODS: Sera of 39 patients infected with different HCV subtypes were examined by a serological genotyping assay (NS-4 IBA), by the widely used INNO-LiPA HCV II, and by a nucleotide sequencing method. RESULTS: The tests performed equally well in terms of HCV subtyping and no different results were obtained. However, the serotyping assay provided the results in less than half the time needed by the other two assays. Significant differences were also observed regarding the 'hands on' times and the costs. The technical equipment which was necessary to perform the assays is significantly reduced using the serological assay. CONCLUSION: Our study demonstrates that the serological test offers the opportunity to determine HCV genotypes and subtypes reliably, fast, easy, and cost effective.

Genotype↗

Serological diagnosis of leishmaniasis: on detecting infection as well as disease.

Serological tests are very frequently used in epidemiological surveys of leishmaniasis and other parasitoses. Their sensitivity and specificity are generally defined with respect to parasitism and disease, rather than infection. The reason is that known positives are those individuals most likely to yield parasites, or who have distinctive clinical signs, and concomitantly high antibody titres. This paper investigates the performance of one serological method, the indirect fluorescent antibody test (IFAT), in detecting Leishmania infantum infection during an intensive 2-year cohort study of dogs in southern France. The results show that sensitivity and specificity with respect to infection can be simultaneously high, but maximum sensitivity is probably < 80%, and lasts for a relatively short period of 2-3 months after a lengthy incubation period. The IFAT gave the incidence of infection as 18-65% in the first year, whereas the best estimate of incidence based on parasite isolation and clinical observation was 72%. But data from the second year suggest that the 72% was itself an underestimate. We argue that, during epidemiological surveys, the IFAT in particular, and serological tests for leishmania in general, will underestimate prevalence, incidence and hence the scale of the control problem. However, there is evidence that tests for canine leishmaniasis employing high threshold titres will identify the most infectious animals, allowing selective treatment or culling of those which contribute disproportionately to transmission.

Animals↗

Differentiation between serological responses to Brucella suis and Yersinia enterocolitica serotype O:9 after natural or experimental infection in pigs.

False-positive serological reactions (FPSR) due to infections with Yersinia enterocolitica serotype Oratio9 (YeOratio9) are a problem in tests for brucellosis. In the present study, FPSR in classical and novel tests for brucellosis following experimental infections of pigs with YeOratio9 were compared with responses of B. suis biovar 2-inoculated pigs. FPSR were limited to 2-9 weeks post-YeOratio9 inoculation, while B. suis-infected pigs were test-positive throughout the 21-week period of investigation. Although YeOratio9-inoculated pigs exhibited FPSR in Brucella tests for a limited period of time, the serological responses in a YeOratio9-purified O-antigen indirect ELISA did not decrease accordingly. Analysis of available cross-sectional serum samples from pig herds naturally infected with YeOratio9 or B. suis biovar 2 confirmed that the observed difference in the duration of the serological responses between the two infections could be used to discriminate between herds infected with B. suis biovar 2 and YeOratio9.

Animals↗

Comparison of rapid office-based serology with formal laboratory-based ELISA testing for diagnosis of Helicobacter pylori gastritis.

Accurate and cost-effective diagnosis of Helicobacter pylori gastritis has taken on major importance. Several serologic tests for the diagnosis of H. pylori infection are commercially available. We compared the performance of the FlexSure HP rapid IgG antibody test with the conventional HM-CAP ELISA to evaluate whether qualitative office-based serology is reliable enough to replace formal laboratory-based testing. We assessed H. pylori status by concordance in 100 consecutive patients with antral biopsy, rapid urease, and 1 microCi[14C]urea breath tests. Both antibody tests had good sensitivity and specificity (>86%). Concordance between the two antibody tests occurred in 87/93 patients (94%). Based on our data, the office-based FlexSure HP performed equally well as the laboratory-based formal ELISA and may be a better choice for initial serologic diagnosis in untreated patients.

Antibodies, Bacterial↗

Serological evidence of alpha herpesvirus infection in sooty mangabeys.

Contact between sooty mangabeys (SMs) and a pigtailed macaque prompted the serological screening of SMs for evidence of infection with B virus. Serological tests detected SM antibodies that reacted with B virus polypeptides. Additional testing was performed with sera from SMs with no previous contact with macaques. Results from these tests indicated that 56% (33/59) of the SMs had antibodies that reacted with B virus and SA8. SM antibodies also reacted with herpesvirus papio 2 and to a lesser extent with human alpha herpesviruses (HSV-1 and HSV-2). There was an age-related increase in the presence of these antibodies in SMs that was consistent with the serological pattern of reactivity observed in other nonhuman primate species infected with alpha herpesviruses. These data suggest that SMs may be a host for a herpesvirus that is antigenically similar to those viruses present in other Old World nonhuman primates.

Age Factors↗

Determinants of human papillomavirus 16 serological conversion and persistence in a population-based cohort of 10 000 women in Costa Rica.

Determinants of human papillomavirus (HPV)-16 serological conversion and persistence were assessed in a population-based cohort of 10 049 women in Guanacaste, Costa Rica. Serologic responses to HPV-16 were measured in 7986 women by VLP-based enzyme-linked immunosorbent assay at both study enrollment (1993/94) and at 5-7 years of follow-up. Seropositive women were defined as >/=5 standard deviations above the mean optical density obtained for studied virgins at enrollment (n=573). Seroconnversion (n=409), persistence (n=675), and clearance (n=541) were defined based on enrollment and follow-up serology measurements. Age-specific distributions revealed that HPV-16 seroconversion was highest among 18- to 24-year-old women, steadily declining with age; HPV-16 seropersistence was lowest in women 65+ years. In age-adjusted multivariate logistic regression models, a 10-fold risk increase for HPV-16 seroconversion was associated with HPV-16 DNA detection at enrollment and follow-up; two-fold risk of seroconversion to HPV-16 was associated with increased numbers of lifetime and recent sexual partners and smoking status. Determinants of HPV-16 seropersistence included a 1.5-fold risk increase associated with having one sexual partner during follow-up, former oral contraceptive use, and a 3-fold risk increase associated with HPV-16 DNA detection at both enrollment and follow-up. Higher HPV-16 viral load at enrollment was associated with seroconversion, and higher antibody titres at enrollment were associated with seropersistence.

Adolescent↗

Optimal serological tests for the detection of Helicobacter pylori infection in the Chinese population.

BACKGROUND: Numerous serological tests for the detection of Helicobacter pylori infection have been developed. However, many perform poorly when evaluated in the Chinese population. AIM: To search for optimal serological tests for the detection of H. pylori infection in Chinese patients. METHODS: Consecutive dyspeptic patients referred for upper endoscopy were recruited. During endoscopy, gastric biopsies were taken for the CLOtest and histological examination. Patients were then given a 13C-urea breath test. Sera were used to test for H. pylori infection, employing three commercial enzyme-linked immunosorbent assay kits (pylori DTect, HP IgG and GAP IgG). Results were compared with the gold standard defined by the CLOtest, histology and 13C-urea breath test. RESULTS: Among the 142 patients (47 male, 95 female; mean age, 49 years) recruited, 81 (57%) were H. pylori-positive, 57 (40%) were H. pylori-negative and four (3%) were defined to be indeterminate. Using a self-defined cut-off value after calculation, the best accuracies for the pylori DTect, HP IgG and GAP IgG tests were 97%, 91% and 80%, respectively. CONCLUSIONS: The pylori DTect test is an optimal serological test for the detection of H. pylori infection in Hong Kong Chinese patients. The HP IgG test may be used as an alternative.

Breath Tests↗

Immunoprecipitation and isoelectric focusing of sheep MHC class I antigens reveal higher complexity than serology.

Biochemical and serological methods were used to characterize sheep MHC class I polymorphism at the product level. The cells of 65 selected animals were subjected to immunoprecipitation and one-dimensional isoelectric focusing (IEF) with two cross-reacting monoclonal antibodies, B1.1G6 and HC-10. We were able to define up to 18 distinct haplotypes in the Latxa breed sample. Most of the locally defined serological specificities were confirmed or subdivided by biochemical typing. As IEF detected antigens not yet defined by serology, it provided us with a guide to producing more specific antisera by appropriate immunizations. Lastly, evidence of expression of two class I loci products was found.

Animals↗

Comparison of Fy(b) status as determined serologically and genetically.

Comparison of the serologically determined phenotype and the genotype of samples as defined by PCR-RFLP indicated a significant percentage of discrepancies on analysis of serologically defined Fy(a + b-) samples. An investigation of these discrepancies has shown that the serologically determined phenotype of the samples varies with respect to the reagent used but that the PCR-RFLP assay gives reliable results for all the samples investigated. The samples which yielded the discrepant results were further investigated and the results indicated weakened expression of the Fyb antigen. This may arise by a method similar to the expression of Fyx although we found the incidence of these samples to be much higher than that reported for Fyx. These results may have important implications for the routine testing of blood donations, particularly in situations where the unit is to be transfused to a patient who has previously formed an anti-Fyb.

Duffy Blood-Group System↗

Report on the Tenth International Platelet Genotyping and Serology Workshop on behalf of the International Society of Blood Transfusion.

BACKGROUND AND OBJECTIVES: The aims of the 10th International Platelet Serology and Genotyping Workshop were to evaluate the proficiency of platelet immunology determinations. MATERIALS AND METHODS: There were 40 participants from 25 countries of four continents. Thirty-eight institutions reported results for genotyping, and 38 institutions reported their serological results. For genotyping, EDTA-anticoagulated whole-blood samples were provided (to allow the inclusion of DNA-separation methodology in the analysis) as well as separated DNA of a and b alleles for human platelet antigen (HPA)-1 to -6. For serological evaluations, sera contained allo- and autoantibodies, and for sensitivity testing a standard freeze-dried sample of HPA-5 antibody. RESULTS: All participants reported HPA-1, -2, -3 and -5 genotyping results; HPA-4 was determined in 29 laboratories and HPA-6 in 21. Results from 16 laboratories were concordant with the majority vote for all allotypes, eight institutions reported one deviation, five laboratories two, and nine laboratories three or more deviations. Twelve institutions had no deviation from the majority vote for HPA antibodies, nine had one, three had two, and 14 had three or more deviations. Most laboratories reported a reactivity of the standard anti-HPA-5b sample with HPA-5b platelets at a dilution of 1:4-1:8. Four laboratories detected anti-Gova in one sample. Seventeen laboratories reported no deviation from the majority vote for pan-reactive platelet antibodies, 12 had one deviation, two had two, and seven had three or more deviations. In addition, seven participants reported antibodies against glycoprotein IV (GPIV), three against glycoprotein V (GPV) and three against CD 109. These results were discussed at a meeting organized jointly with the International Society of Blood Transfusion (ISBT) 2000 Congress. CONCLUSION: The results for pan-reactive antibodies were heterogeneous with most discrepancies from the majority vote. The provision of sufficient samples for many participants is difficult. Based on the results and discussion it is clear that frequent workshops are needed in the future. Therefore, workshops shall be organized regionally, and each region shall participate with one institution in international workshops. The latter are needed to assure international exchange of experience and quality.

Antibody Specificity↗

The utility of serology in diagnosing candidosis in non-neutropenic critically ill patients.

This study was carried out to evaluate the utility of serological tests in the diagnosis of candidal infections in non-neutropenic critically ill patients. A prospective study was carried out in a 10-bed general intensive care unit; all patients with at least one organic sample with Candida spp. were included. A therapeutic-diagnostic algorithm was designed, and patients were treated or not, according to a classification. Blood samples were taken, and serological tests included: antigenaemia detection using two different commercial latex kits (Cand-Tec and Pastorex) and antibody detection by indirect haemagglutination (IHA) and indirect germ tube immunofluorescence (IFA). A total of 56% of antibody tests (IHA 45% and IFA 64%) and 26% of antigen tests (Cand-Tec 36% and Pastorex 17%) were positive. The sensitivity and specificity of these tests with respect to systemic candidosis were 37% and 78%, respectively, for antibodies, and 0% and 90% for antigens. There was statistical significance for mortality and low levels of antibodies; Candida glabrata was detected by IFA and Candida tropicalis by Cand-Tec. Serological tests may help to define the prognosis of these patients and to support the detection of specific Candida species.

Antibodies, Fungal↗

Serological and molecular diagnosis.

The development of serological assays to detect hepatitis B surface antigen (HBsAg) has played a major role in the diagnosis of hepatitis B virus (HBV) infection. With other hepatitis B serological assays, a diagnosis of acute or chronic HBV infection, past infection, or successful vaccination can be determined. However, serological profiles can at times be atypical or ambiguous. Some of these difficulties may be overcome by HBV DNA testing, which may elucidate an individual's true hepatitis B status. In addition, quantification of HBV DNA provides a means of monitoring the effectiveness of antiviral therapy and detecting the early development of antiviral drug resistance. Molecular diagnostics are also being applied to HBV-infected liver tissue. The use of molecular techniques to quantify intrahepatic HBV DNA and other key HBV replicative intermediates may provide additional options for monitoring and predicting treatment efficacy, but such assays require further validation before they can be used routinely.

DNA, Viral↗

Long-term outcome of infrainguinal bypass grafting in patients with serologically proven hypercoagulability.

OBJECTIVE: The purpose of this study was to test the hypothesis that the long-term outcome of infrainguinal bypass grafting in patients with congenital or acquired hypercoagulability is inferior to the results in patients without documented clotting disorders. METHODS: The study was a retrospective analysis of consecutive patients from January 1994 to January 2001. RESULTS: Five hundred eighty-two infrainguinal bypass grafts were created in 456 patients. Indication for surgery was limb-threatening ischemia in 84%; prosthetic conduits were implanted in 38%. Seventy-four grafts were created in 57 patients with one or more serologically proven hypercoagulable states, including heparin-induced platelet aggregation (n = 37), anticardiolipin antibodies (n = 11), lupus anticoagulant (n = 8), protein C or S deficiency (n = 7), antithrombin III deficiency (n = 3), and factor V Leiden mutation (n = 1). Patients with hypercoagulability were younger (63 +/- 2 years versus 69 +/- 1 years; P =.007), more likely to have undergone prior revascularization attempts (38% versus 21%; P =.003), and more likely to have chronic anticoagulation therapy after surgery (46% versus 25%; P =.001). After 5 years (median follow-up, 19 months), patients with hypercoagulability had poorer primary patency (28% +/- 7% versus 35% +/- 5%; P =.004), primary assisted patency (37% +/- 7% versus 45% +/- 6%; P =.0001), secondary patency (41% +/- 7% versus 53% +/- 6%; P =.0001), limb salvage (55% +/- 8% versus 67% +/- 6%; P =.009), and survival (61% +/- 8% versus 74% +/- 4%; P =.02) rates. Multivariate analysis identified only prosthetic conduit choice (P =.0001), hypercoagulability (P =.0003), and limb salvage indication (P =.01) as independent predictors of graft failure. CONCLUSION: Patients with serologically proven hypercoagulability have inferior long-term patency, limb salvage, and survival rates after infrainguinal bypass. The high prevalence rate (13%) of diverse hypercoagulable states in this patient population supports serologic screening, especially in referral practices.

Aged↗

Diagnosis of Helicobacter pylori infection in patients with atrophic gastritis: comparison of histology, 13C-urea breath test, and serology.

BACKGROUND: Atrophic gastritis, a risk factor for gastric cancer, is a late consequence of Helicobacter pylori infection in approximately one-third of the infected patients. It has been suggested that gastric cancer would develop less frequently if H. pylori were eradicated. However, the prevalence of H. pylori infection may be underestimated in patients with atrophic gastritis and intestinal metaplasia if only biopsy-based diagnostic methods are used. METHODS: We compared histology, 13C-urea breath test (13C-UBT), and serology in H. pylori diagnostics in 50 male patients with atrophic corpus gastritis. RESULTS: H. pylori was detected in 15 (30%) patients by histology and in 14 (28%) by 13C-UBT, whereas increased serum antibody levels indicating H. pylori infection were found in 41 (82%) patients (P < 0.0001 between serology and both histology and 13C-UBT). H. pylori infection was associated with atrophic corpus gastritis in 84% of the present patients (in one patient with normal antibody titres H. pylori was defined histologically). CONCLUSIONS: H. pylori infection would have been missed in most patients with atrophic gastritis without the analysis of H. pylori antibodies. Therefore, in patients with atrophic gastritis, the use of serology is encouraged in diagnosing H. pylori infection.

Aged↗

A negative Helicobacter pylori serology test is more reliable for exclusion of premalignant gastric conditions than a negative test for current H. pylori infection: a report on histology and H. pylori detection in the general adult population.

OBJECTIVE: Corpus-dominant gastritis, gastric mucosal atrophy and intestinal metaplasia (IM) associated with Helicobacter pylori infection are all known potential risk markers for the development of gastric cancer. As the accuracy for finding cases at risk in the general population is unknown, we aimed to determine the prevalence of current and/or past H. pylori infection and associated gastric mucosal findings by means of histological survey of a random adult population. MATERIAL AND METHODS: A random Swedish sample (n = 3000, age 20-81 years) was surveyed using a validated gastrointestinal symptom questionnaire with 74% response rate. One-third of the responders were selected at random for esophago-gastro-duodenoscopy with biopsies and H. pylori serology. RESULTS: Of those endoscoped (n = 1000, mean age 53.5, 51% women), 43.0% were H. pylori+ by serology (seropositive), 33.9% had signs of current infection on either histology or culture (gold standard+), and 9.3% were seropositive, but gold standard negative. Corpus atrophy was found in 10% and IM in 13% when gold standard positive, and in a significantly higher number (17% and 21%, respectively) of those with only a serological sign of past infection. Among those who were seronegative, values were 1% and 2%, respectively. Corpus-dominant gastritis was found in 4.1%, all seropositive. CONCLUSION: One-third had an ongoing H. pylori infection, and a further 10% had signs of past infection. Corpus-dominant gastritis was found mostly among the former, while detection of those with corpus atrophy and IM also required a test for past infection. Seronegativity almost excludes precancerous conditions in a screening situation.

Adult↗

Chlamydia pneumoniae is an important cause of community-acquired pneumonia in school-aged children: serological results of a prospective, population-based study.

The aetiology of community-acquired pneumonia in childhood was studied in the total population of 8851 children in the area of 4 municipalities in eastern Finland. All cases of community-acquired pneumonia (n = 201) were registered during a surveillance period of 12 months between September 1, 1981 and August 31, 1982. The diagnosis of pneumonia was verified radiologically in all identified cases. The diagnosis of chlamydial infection was based on an antibody response measured by complement fixation (CF), by enzyme immunoassay (EIA; IgG or IgM) or by microimmunofluorescence (MIF; IgG or IgM), and the diagnosis of mycoplasmal infection on CF alone. In total, 29 cases of Chlamydia sp. infection were diagnosed; 20 were caused by Chlamydia pneumoniae. Thus, C. pneumoniae was an aetiological agent in 10%, of the 201 pneumonia cases: the proportion was 9% for children aged 5-9 y and 31% for those aged 10 y or more. In the study population, the total incidence of C. pneumoniae pneumonia was 2.3/1000/y. Mycoplasma pneumoniae serology (CF) was positive in 44 patients (22%); the total incidence of M. pneumoniae pneumonia was 5.0/1000/y. Serological evidence of both Chlamydiae and M. pneumoniae was detected in 9 (41%) patients. Our results indicate that C. pneumoniae is an important cause of community-acquired pneumonia in school-aged children. Diagnostic serological response to Chlamydia species or M. pneumoniae was found in 42% of pneumonia patients between 5 and 9 y of age and in 67% of patients aged 10 y or more. Thus, we suggest that macrolides should be considered as an empirical antimicrobial treatment for community-acquired pneumonia, especially in school-aged outpatients.

Adolescent↗

Late onset of serologic positive titers in a patient with Parinaud's oculoglandular syndrome.

PURPOSE: To report a case of Parinaud's oculoglandular syndrome (POS) in which, despite the initially negative, a late onset of serologic positive titers was observed. METHODS: Case report. RESULTS: A 65-year-old man was examined for a foreign body in the left eye associated with granulomatous nodule on the palpebral conjuctiva, chemosis and a swollen preauricular lymph node. Despite the initially negative serologic titers to Bartonella henselae, a clinical diagnosis of POS was made and treatment was instituted initially with oral ciprofloxacin (500 mg twice a day). Three weeks later, serologic positive titers were found which confirmed the diagnosis of POS. CONCLUSION: Even though the improvements in diagnostic testing such as indirect immunofluorescence antibody, negative titles could be initially observed, confusing the diagnostic process. Other techniques (such as polymerase chain reaction (PCR) testing) should be also taken into consideration in cases with increased clinical suspicious of B. henselae indections.

Aged↗