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Serological diagnosis and follow-up of asymptomatic and acute Q fever infections.

During an outbreak of Q fever at a farmer's market in Soest (North Rhine-Westphalia, Germany) in 2003, we examined 263 serum samples of presumably infected persons for Q fever antibodies. One hundred and seventy-one of these patients were tested positive for acute Q fever infection. Furthermore, 29 persons of certain risk groups like pregnant women (n=11) or patients with valvular heart disease (n=18) were examined. Among these, in four pregnant women and two heart patients an acute but asymptomatic infection could be diagnosed. With 30 patients we performed a serological follow-up for 8-60 weeks. In our study, phase 2 (PH2)-IgM antibodies as a marker for acute infection were present in all 30 patients 3-4 weeks after onset of clinical signs and disappeared 3-4 months later. Six weeks to three months after clinical manifestation, all patients developed PH1-IgG antibodies in low levels with no clinical signs of chronic Q fever. Three patients, including one pregnant woman showed high-level titres and were treated for chronic Q fever. Eleven patients with low PH1-IgG antibodies and all three patients with high titres developed IgA antibodies from 10 weeks after clinical manifestation; therefore PH1-IgA cannot be used as the only serological marker for chronic Q fever. Chronic infections were indicated only by a continuous increase of PH1 antibodies and a high persistence of PH2-IgG. We therefore conclude that the exclusion of chronic Q fever infection by a single serological examination cannot be done. At least three consecutive tests should be performed, that is 3, 6, and 9 months after initial Q fever infection.

Acute Disease↗

Chlamydophila (Chlamydia) pneumoniae serology and asthma in adults: a longitudinal analysis.

BACKGROUND: Many cross-sectional studies have found an association between Chlamydophila pneumoniae infection and asthma, and a possible causative role of C pneumoniae infection in asthma pathogenesis has been proposed. No longitudinal studies have been undertaken to estimate the effect on asthma incidence of previous or chronic infection. OBJECTIVE: We evaluated whether serological findings suggestive for recent or chronic C pneumoniae infection affect asthma risk or lung function during follow-up. METHODS: We followed a population-based adult cohort for 15 years and made a clinical evaluation of persons with new persistent asthma (n = 83) and matched controls (n = 162). Serological testing was performed by microimmunofluorescence and enzyme immunoassay from both baseline and follow-up samples. RESULTS: Subjects with serologically diagnosed recent or chronic C pneumoniae infection did not run a higher risk of new asthma. An increased risk was found in subjects with allergic rhinitis, low lung function, history of smoking, and positive family background of asthma or allergy. However, chronic C pneumoniae infection was found to accelerate the loss of lung function significantly in subjects who contracted new nonatopic asthma (median change in FEV(1), 89.6 vs 55.9 mL/y; P = .032). CONCLUSION: Chronic C pneumoniae infection promotes the development of airflow limitation in adults with nonatopic asthma. However, our results indicate that at the population level, any possible effect of C pneumoniae infection on asthma incidence is of minor significance.

Adult↗

Epidemiology of HHV-8 infection in HIV-positive patients with and without Kaposi sarcoma: diagnostic relevance of serology and PCR.

BACKGROUND: Kaposi's sarcoma (KS) remains the most common neoplasm in HIV-infected patients. Human herpesvirus 8 (HHV-8) infection is etiologically associated with KS. Diagnostic procedures with regard to HHV-8 infection are not routinely performed in HIV-infected patients; diagnostic and prognostic value of HHV-8 serology or PCR are unknown in this setting. Epidemiological data concerning HHV-8 infection of HIV-infected patients in Germany are rare. OBJECTIVES: To assess prevalence of HHV-8 infection in a cohort of HIV-infected patients with and without KS in Germany and to correlate this to manifestations and clinical course of KS. STUDY DESIGN: HHV-8 serology was performed in 483 patients in routine care for HIV-infection in northern Germany. HHV-8 DNA was analyzed by PCR in peripheral blood mononuclear cells (PBMC) of 293 patients; in a subgroup multiple samples were analyzed. History and manifestations of KS were recorded. RESULTS: HHV-8 antibodies were detected using IFT in 91% of 33 patients with KS and 52% of 398 patients without KS. In 36 of 293 (12.3%) patients HHV-8 DNA was detected in PBMC. In general, HHV-8 DNA was not continuously detected when multiple samples from the same patient were analyzed. Patients with KS history were more likely to be PCR positive than those without (45.5% versus 7.8%). In patients with active KS HHV-8 DNA was detected more frequently than in patients with disease remission. HHV-8 DNA was not detected in serologically negative patients. However, three patients with KS history in full remission for several years were seronegative. CONCLUSIONS: HIV-infected patients were frequently found to be positive for HHV-8 antibodies. The number of patients positive for viral DNA in PBMC was much smaller. Single PCR-examinations were of little value for prognosis, but repeated detection of HHV-8 DNA represents an increased risk of disease activity.

Base Sequence↗

Routine diagnostics of Lawsonia intracellularis performed by PCR, serological and post mortem examination, with special emphasis on sample preparation methods for PCR.

The aim of this study was to find suitable and reliable tools for demonstrating Lawsonia intracellularis in routine clinical diagnosis. Firstly, a method to prepare tissue samples before a polymerase chain reaction (PCR) was evaluated in pigs submitted for necropsy. Secondly, seven different faecal preparation methods and four different DNA polymerases were tested in single or nested PCR, with co-amplification of a mimic molecule. Thirdly, in selected pigs submitted for necropsy, tissue and faecal samples were examined histopathologically and by PCR, and blood samples were analysed serologically. Detection of L. intracellularis in tissue preparations by PCR showed good specificity and correlated to lesions found at necropsy. The sensitivity in spiked tissue samples was 10(1)-10(2) mimic molecules per tube. In faecal samples, nested PCR on boiled lysate gave the best result with a sensitivity of 10(2)-10(3) mimic molecules per reaction tube. However, because of the time-consuming procedure and the increased risk for contamination, a commercially available kit was preferred for routine diagnoses, despite a somewhat lower detection rate in subclinically infected pigs. In a few cases, the serological results differed from those obtained by PCR and by necropsy but the reason for this is not clear. This study indicates that the best method for diagnosis of acute enteritis in growers is PCR on faecal or tissue samples. To determine the presence of the bacteria in a herd, serology or repeated faecal sampling for PCR from target animals, or both, should be used.

Animals↗

Serological diagnosis of canine alveolar echinococcosis.

Canine alveolar echinococcosis (AE) is a rare liver disease caused by the larval stage (metacestode) of E. multilocularis. Its diagnosis so far is rather difficult and invasive. Imaging techniques for the diagnosis have been described recently and in this study, the serological diagnosis of canine AE was evaluated in 30 dogs with confirmed AE using ELISA based on seven different antigens (E. multilocularis-antigens: Em2G11, EmII/3-10, protoscolex (EmP), excretory/secretory and adult integument (EmAd/I); E. granulosus antigens: hydatid fluid (EgHF) and antigen B (EgAgB)) and by the detection of circulating Em2G11-antigen. EmII/3-10 and Em2G11-based tests detected AE in 50 and 53%. The combined serological investigation for specific circulating Em2G11-antigen and specific antibodies against this antigen showed a sensitivity of 77%. This strategy is useful in animals with large parasite masses and ascites. The highest sensitivities for AE were obtained by the EmAd/I and the EmP antigens (97 and 93% sensitivity) with high specificities in the control group of 76 dogs (100 and 98.7%, respectively). Heterologous E. granulosus antigens (EgHF and EgAgB) showed low sensitivities (43 and 50%, respectively) with high numbers of unspecific reactions (>16%) in the control group. In the follow-up of four dogs, the serological reaction patterns against the antigens Em2G11, EmP and EmAd/I showed some correlations with the development of the parasite mass. Intestinal infections cannot be differentiated from AE by most antigens (except EmII/3-10) and have to be excluded by the absence of coproantigens or DNA of E. multilocularis in faecal samples.

Animals↗

Mooren ulcer in South India: serology and clinical risk factors.

PURPOSE: To investigate the rate of undiagnosed rheumatologic diseases and hepatitis C infection among patients with the clinical diagnosis of Mooren ulcer seen at Aravind Eye Hospital, Madurai, South India. METHODS: Twenty-one patients with the clinical diagnosis of Mooren ulcer and 44 control patients underwent a complete ophthalmic history and examination, as well as serologic testing for antinuclear antibodies, rheumatoid factor, antineutrophil cytoplasmic antibodies, herpes simplex virus 1 antibodies, and hepatitis C virus antibodies. RESULTS: There were no statistically significant differences in the rates of seropositivity for antinuclear antibodies, rheumatoid factor, antineutrophil cytoplasmic antibodies, herpes simplex virus 1 antibodies, and hepatitis C virus antibodies between patients with Mooren ulcer and control patients. Two patients with Mooren ulcer and four control patients were found to have a rheumatoid factor titer of greater than 1:20. One of the control patients, but none of the patients with Mooren ulcer, was found to have serologic evidence of hepatitis C infection. A history of corneal trauma, surgery, or infection was reported by 68% of patients with Mooren ulcer, compared with 20% of control patients (P < .001). Among patients with Mooren ulcer, bilateral disease occurred in 37% of patients, visual acuity was reduced to light perception in 15% of eyes, and perforation occurred in 19% of eyes. CONCLUSIONS: Nineteen (90%) of 21 patients with the clinical diagnosis of Mooren ulcer were found to have no evidence of an underlying rheumatologic disease by history, examination, or serologic testing, and none was seropositive for hepatitis C. However, patients with Mooren ulcer were more likely than control patients to report a history of corneal trauma, surgery, or infection.

Adolescent↗

Diagnosis of gastritis by means of a combination of serological analyses.

BACKGROUND: Gastroscopy and examination of biopsy is normally required for diagnosis of gastritis. This is costly and inconvenient for the patient, and there is a need for a simple pregastroscopic screening method to reduce the endoscopy workload. Our aim was to develop a serological screening test for gastritis. METHODS: Sera from subjects examined with gastroscopy and biopsy were analyzed for H,K-ATPase antibodies, Helicobacter pylori antibodies and pepsinogen I. The diagnoses were normal gastric mucosa (n=50), duodenal ulcer (n=53) and atrophic corpus gastritis, with (n=50) or without pernicious anemia (n=46). RESULTS: An evaluation scheme was constructed to optimize the diagnostic agreement between serology and gastric mucosal morphology. The sensitivity to detect gastritis was 98% (146/149) (95% CI 94-100%) and the specificity 84% (42/50) (95% CI 71-93%). Additional sera from 483 subjects from the general population were analyzed. There was a good agreement between serology and gastric mucosal morphology. CONCLUSIONS: Assays of multiple serum analytes are useful for the initial screening of gastritis. They are complementary to upper gastroscopy by identification of subjects with a normal gastric mucosa, those who qualify for eradication of H. pylori, and those who have developed atrophy and are at risk of developing malignancy and, therefore, require gastroscopic examination.

Adult↗

[Serological testing and prevalence of human immunodeficiency, hepatitis B and C viruses infections amongst acute psychiatric inpatients].

BACKGROUND AND OBJECTIVE: The request for serological tests for HIV, HBV and HCV is usual amongst acute psychiatric inpatients. Nevertheless, the seroprevalence and the repercussion of a positive result have been scarcely studied. PATIENTS AND METHOD: Serological requests for these viruses were retrospectively analyzed in a sample of 332 psychiatric inpatients. RESULTS: Serological tests were requested by 65.3% patients. The prevalence of any seropositivity was 7.3%: 1.4% were infected by HIV-I, 3.2% had been in contact with HBV but none had an active infection, and 5.1% were infected by HCV. The presence of a positive result did not change the psychiatric therapeutic approach in any case. CONCLUSIONS: The low prevalence of seropositivity suggests that such tests should be performed selectively in patients who present risk factors or atypical psychiatric symptoms.

Acute Disease↗

Comparison of the polymerase chain reaction and serology for the diagnosis of HTLV-I infection.

Human T-lymphotropic virus type I (HTLV-I) is associated with tropical spastic paraparesis (TSP) and adult T-cell leukaemia/ lymphoma (ATL). HTLV-I seroconversion may not take place for many years after infection and some patients develop relatively low concentrations of antibodies that are difficult to detect by means of conventional assays. Some serologic tests also yield high rates of false-positive results. We therefore decided to determine whether the polymerase chain reaction (PCR) might be more sensitive than serology for the diagnosis of HTLV-I infection. Samples of serum and of peripheral blood mononuclear cells were obtained from 50 patients with spastic myelopathy and nine with T-cell neoplasia. The serum samples of serum were tested for the presence of HTLV-I antibodies by means of an enzyme-linked immunosorbent assay (ELISA). Positive results were confirmed by Western blot (WB) assay. DNA was extracted from the peripheral blood mononuclear cells and a PCR performed by use of two primer pairs from the env and pol regions of the proviral genome. Seven of the 50 patients with myelopathy and none of those with T-cell neoplasia had detectable antibodies to HTLV-I and all seven were PCR positive. One patient with an indeterminate WB result and one who was HTLV-I antibody-negative, both with myelopathy, were also PCR-positive. In this study, PCR was found to be more sensitive than serology for the diagnosis of HTLV-I infection. PCR should therefore be considered for selected HTLV-I antibody-negative patients with unexplained spastic myelopathy or T-cell neoplasia.

Adolescent↗

Evaluation of humoral response to tumor antigens using recombinant expression-based serological mini-arrays (SMARTA).

Screening of expression cDNA libraries derived from human neoplasms with autologous sera (SEREX) is an established method for defining antigens immunogenic in individual cancer patients. Although the majority of SEREX-derived cDNA clones encode autoantigens, some of them represent shared cancer antigens with cancer-related serological profiles. Routine evaluation of multiple SEREX-derived clones in serological assays using panels of allogeneic sera from cancer patients is an important step towards defining disease parameters of diagnostic and prognostic significance. Here we show how the seroreactivity of multiple SEREX-derived antigens can be simultaneously evaluated using a rapid semi-quantitative protocol of allogeneic screening, which we call SMARTA (serological mini-arrays of recombinant tumor antigens).

Antibodies, Neoplasm↗

Prevalence of Chlamydia pneumoniae in the atherosclerotic plaque of patients with unstable angina and its relation with serology.

BACKGROUND: Chlamydia pneumoniae has been associated with coronary artery disease by both seroepidemiological studies, and by direct detection of the micro-organism in atherosclerotic lesions. This bacteria could play a potential role in the development of acute coronary events. We examined coronary arteries from patients with unstable angina in order to verify an endovascular presence of C. pneumoniae, and to determine if there is any relationship between serology of acute infection by this pathogen and its presence inside the atherosclerotic plaque of these patients. METHODS: We analysed a total of 76 atherosclerotic plaques obtained from 45 patients who underwent coronary artery bypass surgery. In all patients unstable angina was present within the prior 3 weeks. The presence of C. pneumoniae in the plaque was determined by nested polymerase chain reaction (PCR). Antichlamydial immunoglobulin G (IgG), A (IgA) and M (IgM) was examined by microimmunofluorescence and compared to the PCR result. FINDINGS: DNA of C. pneumoniae was detected in 57 (75%) of 76 atherosclerotic lesions. In most cases (74/76: 97%) a positive IgA, IgM or IgG result was seen. Seven (12%) and 54 (94%) of the 57 PCR positive plaques came from patients with a positive IgM and IgA result, respectively. There was no statistical significant difference between PCR positive and PCR negative plaques in patients with a positive or negative serological result. Clinical characteristics were similarly distributed in patients with and without infected lesions. INTERPRETATION: C. pneumoniae organisms are frequently found in the atherosclerotic lesions of patients undergoing coronary surgery for unstable angina. Neither serological results of acute or recent infection by C. pneumoniae nor clinical characteristics are useful in predicting the individual risk of harbouring C. pneumoniae in the coronary lesions of patients with unstable angina.

Adult↗

[Failure of rapid serology for Helicobacter pylori as diagnostic method in primary care consultation].

OBJECTIVE: To evaluate the validity of a rapid serology test for diagnosing Helicobacter pylori infection in our area. DESIGN: Longitudinal, prospective study. SETTING: Urban primary care centre. MEASUREMENTS: As reference tests for defining the presence/absence of Helicobacter pylori, histology and the urease gastric mucous membrane test (Jatrox(R) test) were used. Both tests can be either positive or negative, without intermediate values. RESULTS: 47 consecutive patients were included. Histology (confirmed with the urease test) showed infection in 38 cases (80.85% prevalence). The results of rapid serology (95% CI) were: sensitivity 0.66 (0.51-0.81), specificity 1 (1-1), positive predictive value 1 (1-1), and negative predictive value 0.41 (0.2-0-61). CONCLUSIONS: In a population with high prevalence of infection, a positive in the rapid serology test confirms infection. However, a negative does not discount infection, which significantly reduces the test's overall diagnostic value. This confirms in the primary care context the findings of hospital-based studies.

Adult↗

Assessment of prescription practices for serological tests for viral hepatitis B and C in the Greater Parisian area in 2002.

OBJECTIVES: The aim of this study was to ascertain the reasons for the prescription of serological tests for viral hepatitis B (HBV) and C (HCV) in the Greater Parisian area, and to assess standards of prescription with respect to current guidelines. PATIENTS AND METHODS: The population studied comprised patients affiliated to the three main health insurance schemes in the Greater Parisian area, for whom at least one serological test for HBV or HCV was reimbursed on May 14th or 15th, 2002. Data was collected from prescribing and laboratory heads. RESULTS: The sample consisted of 1 046 prescription orders for HBV and/or HCV tests. The mean age of patients was 39 years, and 68% were females. The main medical indications declared by the prescribing physicians were: screening for HBV or HCV (40%), tests for pregnant women (19%), suspected hepatitis B or C (13%), pre- and post-vaccination tests for HBV (7%), medically assisted procreation (6%), follow-up of diagnosed chronic hepatitis (4%). Assessment of the standards of prescription orders showed a lack of compliance with guidelines for 71% of HBV tests, and 56% of HCV tests. CONCLUSIONS: The implementation of guidelines for the prescription of serological tests for HBV and HCV needs to be improved in clinical practice.

Adolescent↗

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↗

Insufficient diagnostic accuracy of imported serological kits for Helicobacter pylori infection in Japanese population.

Although there are many reports of the high diagnostic accuracy of commercially available serologic kits for Helicobacter pylori infection in Western countries, they rarely has been investigated in oriental population. Accordingly we examined their usefulness in 492 Japanese patients with dyspeptic symptoms. Diagnostic accuracy of 4 imported serologic kits (HEL-p TEST, HM CAP, G.A.P IgG, Helico G2) was investigated using the (13)C-urea breath test as the gold standard. When intermediate results were excluded, the sensitivity, specificity and accuracy of these serologic tests ranged from 88.6% to 97.8%, 67.9% to 85.9%, and 87.9% to 91.4%, respectively, which were comparable with reported median accuracy in the Western population. However, there were many intermediate results in these tests, ranging from 5.3% to 23.0%. Their usefulness seemed to be limited in our patient population because of the large number of intermediate results.

Adult↗

Accurate diagnosis of Helicobacter pylori. Serologic testing.

Serologic testing is a useful noninvasive testing strategy for H. pylori. It is particularly useful in areas where the prevalence of H. pylori is high and inexpensive point-of-contact fingerprick tests are used. Sensitive tests are valuable ways of excluding H. pylori infection and can be used in conjunction with a direct test (urease histology culture or breath test) to confirm absence of H. pylori if the two methods are concordant. Serologic testing is more definitive and differentiating if the antigenic epitopes of H. pylori can be differentiated based on the antigenic epitopes that specifically associate with gastric cancer, peptic ulcer, and nonulcer dyspepsia. A study by Kawahara's group reported that Hsp 60 may be involved in the development of mucosa-associated lymphoid tissue based on ELISA. The idea of differentiating antigens for H. pylori may open a new area for use of serologic testing in the diagnostic approach of H. pylori infections.

Antibodies, Bacterial↗

Serologic markers for Chlamydia pneumoniae in asthma.

BACKGROUND: Chlamydia pneumoniae infection has been reported as a possible etiologic agent in asthma, which in primary care settings often appears to be initiated by acute respiratory infections. OBJECTIVE: To determine if serologic markers for C. pneumoniae are associated with adult asthma that first became symptomatic after an acute respiratory illness (asthma associated with infection: AAWI). METHODS: Serum samples from 164 primary care outpatients, mean age 44 years, (68 with AAWI; 36 with atopic, occupational or exercise-induced asthma (non-AAWI); 16 nonasthmatic patients with acute bronchitis; and 44 asymptomatic nonasthmatic controls) were tested for the presence of C. pneumoniae-specific IgG and IgA antibodies. Levels of chlamydial heat shock protein 60 (CHSP60) antibody were also measured. Those positive for CHSP60 were tested for C. pneumoniae-specific IgE antibodies by immunoblotting. RESULTS: Statistically significant differences in IgG and IgA seroreactivity were noted between groups: acute bronchitis and AAWI had the highest levels (93% to 94% IgG seroreactivity, 69% to 75% IgA seroreactivity) whereas non-AAWI and asymptomatic controls had the lowest levels (61% to 84% IgG seroreactivity, 31% to 43% IgA seroreactivity, P < .02 after adjustment for age, sex and smoking). CHSP60 antibodies were significantly more prevalent in AAWI than in non-AAWI (19% versus 3%, P = .02). IgE antibodies against C. pneumoniae 60, 62, and/or 70 kD antigens were detected in 5 of 13 CHSP60 positive AAWI patients. Persistent IgG, IgA, and CHSP60 seroreactivities were noted in all seropositive asthma patients with serial serum samples. CONCLUSIONS: Serologic markers of C. pneumoniae infection were associated with acute bronchitis and with asthma that first became symptomatic following respiratory illness. Serologic responses to C. pneumoniae may be useful in the classification and diagnosis of asthma.

Adult↗

FIV, FeLV, and FIPV: interpretation and misinterpretation of serological test results.

Serological testing is a common method of diagnosis of felina viral infections, including feline immunodeficiency virus (FIV), feline leukemia virus (FeLV), and feline infectious peritonitis virus (FIPV). Infections with these viruses can be difficult to diagnose by clinical signs alone and are sometimes clinically inapparent for months after initial exposure. Serological testing to confirm a tentative diagnosis or as a screening tool for infection can be invaluable. However, serological tests must be used only with a thorough understanding of the mechanisms and abilities of the tests, and with recognition of their potential inadequacies and misinterpretations. This report summarizes the assays available for FIV, FeLV, and FIPV, and discusses merits and pitfalls associated with each test.

Animals↗