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Accuracy of the serological ELISA test compared with the polymerase chain reaction for the diagnosis of cytomegalovirus infection in pregnancy.

CONTEXT: The most frequently used methods for detecting antibodies are the indirect immunofluorescence test and the enzymatic immunoassay (ELISA). The polymerase chain reaction is a molecular biology technique in which the production of large amounts of specific DNA fragments is induced from very low concentrations of complex substrates aloowing the detection of very low amounts of viral particles. OBJECTIVE: To assess the accuracy of serological/ELISA tests in comparison with the polymerase chain reaction in maternal blood to diagnose cytomegalovirus infection. DESIGN: A descriptive study was performed. SETTING: High-risk outpatient clinic of Campinas University (Unicamp). PARTICIPANTS: We selected 243 pregnant women. All of them had been indicated for blood sampling because of suspicions of cytomegalovirus infection and also because of other infections. MAIN MEASUREMENTS: The group was tested for cytomegalovirus. Serological tests were run and compared to the polymerase chain reaction, which was considered to be the gold standard. Status analyses were done using Fisher's exact test, via the SAS software. RESULTS: The previous cytomegalovirus infection rate was 94.6%. The main reasons for inclusion in the study were fetal nervous system malformation (25.5%), maternal toxoplasmosis (25.5%) and Rh isoimmunization (14.8%). Only two women were included because of positive serological immunoglobulin M test for cytomegalovirus. The sensitivity and specificity of the serological tests were 94% and 6% for immunoglobulin G. CONCLUSION: Serological tests had lower sensitivity in comparison with the polymerase chain reaction test when diagnosing cytomegalovirus infection. The consequences of positive polymerase chain reaction and negative immunoglobulin M in women remain unknown.

Adult↗

How reliable are the 14C-urea breath test and specific serology for the detection of gastric Campylobacter?

Detection of gastric Campylobacter by the 14C-urea breath test and serology were correlated to biopsy culture in 25 unselected outpatients referred for gastroscopy. All the 17 culture-positive patients had positive 14C-urea breath test, and 16 had positive serology. Of eight culture-negative patients, six patients had negative breath test and seven negative serology. A high degree of reproducibility was found when two subsequent breath tests were performed in 11 healthy volunteers. The breath test values obtained at 10 min showed a strong correlation (r = 0.97, p less than 0.001) to the accumulated values within 30 min. Breath sampling once, 10 min after intake of 2.5 microCi 14C-urea, seems sufficient for the detection of gastric Campylobacter. The 14C-urea breath test correlates well with biopsy culture and provides a sensitive tool for the detection of gastric Campylobacter. Serology also corresponds well with biopsy culture and should provide a useful tool for epidemiologic studies.

Adult↗

Febrile illnesses resulting in hospital admission: a bacteriological and serological study in Jakarta, Indonesia.

A study of 741 Indonesian patients with fever was carried out in order to determine what serious febrile illnesses are prevalent in Jakarta. All patients were hospitalized primarily because of fever and were studied by bacteriological and serological methods. Bacteremia due to Salmonella typhi (150 cases), S. enteritidis (36 cases), or both (2 cases) was common in both children and adults. One S. enteritidis isolate was chloramphenicol resistant. Serological evidence of Salmonella infection was found in 130 additional cases without bacteremia. Serological evidence of arbovirus infection (94 cases) was common in children. Malaria was found in 12 adults, most of whom were probably infected outside Jakarta. Little serological evidence was found for rickettsial, leptospiral, Brucella, Toxoplasma gondii or a number of other infections. Clinical signs and symptoms in the febrile patients studed were generally nonspecific, and laboratory results reported were very helpful in establishing more accurate diagnoses.

Arbovirus Infections↗

Serologic evidence of previous Campylobacter jejuni infection in patients with the Guillain-Barré syndrome.

OBJECTIVE: To determine if patients with the Guillain-Barré syndrome are likely to have had Campylobacter jejuni infection before onset of neurologic symptoms. DESIGN: A case-control study. SETTING: Several university medical centers. PATIENTS: Case patients met clinical criteria for the Guillain-Barré syndrome between 1983 and 1990 and had a serum sample collected and frozen within 3 weeks after onset of neurologic symptoms (n = 118). Disease controls were patients with other neurologic illnesses (n = 56); healthy controls were hospital employees or healthy family members of patients (n = 47). MEASUREMENTS: Serum IgA, IgG, and IgM antibodies to C. jejuni were determined by enzyme-linked immunosorbent assays. Assays were done in a blinded manner. RESULTS: Optical density ratios > or = 2 in two or more immunoglobulin classes were seen in 43 (36%) of patients with the Guillain-Barré syndrome and in 10 (10%) of controls (odds ratio, 5.3; 95% CI, 2.4 to 12.5; P < 0.001). Increasing the optical density ratio or the number of immunoglobulin classes necessary to yield a positive result increased the strength of the association. The number of patients with the Guillain-Barré syndrome who had positive serologic responses was greatest from September to November (P = 0.02). Male patients were three times more likely to have serologic evidence of C. jejuni infection (P = 0.009); the proportion of patients with the syndrome who had a positive serologic response increased with age. CONCLUSIONS: Patients with the Guillain-Barré syndrome are more likely than controls to have serologic evidence of C. jejuni infection in the weeks before onset of neurologic symptoms. Campylobacter jejuni may play a role in the initiation of the Guillain-Barré syndrome in many patients.

Adolescent↗

[Influence of standardization upon the results of serological analysis of toxoplasmosis (author's transl)].

It is the purpose of this paper to determine whether the decline of the serologically verified acute Toxoplasma gondii infections observed since the late sixties and found in literature has been caused by epidemiological factors or whether the standardization of serological methods (10) introducted in 1966/66 for the determination of antibodies specific to Toxoplasma gondii has been responsible. The paper is based on material of the prospective study, "Pregnancy and child development" supported by the Deutsche Forschungsgemeinschaft (DFG). Mainly six laboratories did the serological analysis for antibodies against T. gondii from 1964 to 1970. The following results were obtained: 1. Laboratories using the standardization observed a decline of the highly positive dilution starting in 1968 (Fig. 1, Table 1). 2. The proportion of the samples positive in the dye test did not change (Fig. 1). 3. The proportion of sera reacting in the complement fixation test (CFT) with a dilution of 1:5 increased. But after introduction of the standardization more sera were analyzed by both the dye and complement fixation tests than before (Table 2, Fig. 2). 4. The proportion of sera found positive by the complement fixation test increased slightly up to 1968 (Fig. 3). 5. Differences in the age structure (Fig. 4) or dietary habits of the group (Fig. 5) did not cause the changes in titer. 6. Therefore the standardization was introduced a considerable proportion of the sera showing dye test results of more than 1:1000 reacted negatively to the complement fixation test (Table 4). These cases were probably not acute infections. 7. When the standardization was introduced the difference between results of first and second analysis decreased for sera with titers smaller then 1:256 (increased of reliability) (Table 5, Fig. 7). 8. Complement fixation titer show an analogous behavior. (Table 7 and 8). The results detained above show that the decrease, observed since 1969, of the cases of acute Toxoplasma infections verified by serological methods is not caused by epidemiological but rather by methodical factors.

Adult↗

Reducing the endoscopic workload: does serological testing for Helicobacter pylori help?

OBJECTIVE: Serological screening for Helicobacter pylori (H. pylori) in young (< or = 45 years) dyspeptic patients has been used to avoid oesophago-gastro-duodenoscopy (OGD). We used serology to identify seronegative and seropositive patients without sinister symptoms and combined approaches of avoiding OGD in both groups. We aimed to determine the reduction of OGD in this group. DESIGN: Prospective study on the treatment of 232 patients with dyspepsia. SETTING: Six hundred and fifty bed district general hospital serving rural Northamptonshire, UK. INTERVENTIONS: Two hundred and thirty-two patients referred by local general practitioners for OGD were offered serology. Symptom severity was scored using a questionnaire. One hundred and eleven seronegative patients received symptomatic treatment, 105 seropositive patients received triple therapy for 1 week. Sixteen patients with equivocal results were offered OGD. Patients were followed up after 6 months. MAIN OUTCOME MEASURES: Severity of dyspepsia symptoms and proportion of patients returning for OGD. RESULTS: Fifteen equivocal patients underwent OGD, one refused. Forty-six patients (33 seronegative, 13 seropositive) had persisting symptoms and underwent OGD. Mean symptom severity was reduced significantly in equivocal (P<0.01), seronegative (P<0.001) and seropositive (P<0.001) patients. Fewer seronegative patients were symptom-free at follow up compared to seropositive patients (n = 15 (16%) vs n = 48 (51%); P<0.001), 171 patients avoided OGD, a 74% reduction. CONCLUSIONS: Use of H. pylori serology in the management of young dyspeptic patients without sinister symptoms can reduce the OGD workload by 74%, decreasing the length of time that older patients, who are at greater risk of malignant disease, may have to wait for OGD.

Adolescent↗

[A comparative study of serological, monoclonal antibody and DNA typing in identifying HLA-A].

OBJECTIVE: A double-blind study was carried out to evaluate the accuracy and reliability of PCR-SSP assay in comparison with serology and monoclonal antibody(mAb) typing in identifying HLA-A alleles in Southern Chinese population. METHODS: A total of 296 samples were entered into the study, including 143 unrelated kidney donors and 153 recipients. HLA-A typing was performed by standard two-stage microlymphocytotoxicity assay, one-step mAb typing and PCR-SSP typing. RESULTS: All samples were successfully typed by PCR-SSP. Reproducibility was 100%. The results were confirmed by a panel of standard DNAs and a double-blind typing of UCLA tissue typing lab. However, mAb typing(for Asian) in 149 samples showed 2.7% misassignment including 1 antigen being incorrectly interperted and 3 of 26 "blanks" turning out to be definable alleles by DNA typing. Serological discrepancy rate was 15.6% in 147 samples consisting of 8 antigens being incorrectly interpreted, 13 "blanks" turning out to be definable alleles and 2 heterozygotes turning out to be homozygotes by DNA typing. CONCLUSION: HLA-A typing by PCR-SSP proved to be a rapid and accurate technique, suitable for clinical application with a greater precision than serology. In large scale screening, mAb typing (for Asian) is recommended. Antigens of "blanks" or "difficult" by serology or mAb typing should be retyped by DNA typing.

Alleles↗

Empiric treatment based on Helicobacter pylori serology cannot substitute for early endoscopy in the management of dyspeptic rural black Africans.

BACKGROUND: Evidence that chronic gastric Helicobacter pylori (HP) infection is an aetiological factor in dyspepsia, peptic ulcer disease, gastric carcinoma and lymphoma has led to the suggestion that all serologically positive dyspeptic patients should be treated empirically with antibiotics to eradicate the infection, without endoscopic diagnosis. The following study was performed to determine whether such a policy would prove to be of benefit in rural Africa, where endoscopic facilities are lacking and infection rates high. METHODS: Four district clinics were visited and 97 consecutive patients with persistent upper gastro-intestinal symptoms studied. After history-taking and physical examination, a blood sample was taken for HP serology (IgG anti-HP EIA) and endoscopy was performed. RESULTS: In comparison with similar studies in westernised countries HP was considerably more common (80%), and similar to that reported for the background population (83-86%), but peptic ulceration (17%) and gastric cancer (1%) were not. HP status and antibody levels failed to predict the presence of serious disease; patients with 'alarm' signs (78%), cancer (78%) and peptic ulcers (81%) had similar seropositivity rates to patients with non-ulcer dyspepsia (81%). Interestingly, many patients with distal oesophagitis were seronegative (40%). Haemoglobin concentrations and nutritional status were similar in HP-positive and negative patients. On the basis of published decision analysis strategies, empiric treatment of HP-positive patients with uncomplicated dyspepsia could be expected to produce symptomatic relief in 50% of cases, but would have delayed the diagnosis of 3 cases of cancer if patients over the age of 45 were included. CONCLUSION: The lack of association between HP serology and upper gastro-intestinal disease indicates that serological investigation cannot substitute for endoscopy in the management of black Africans with dyspepsia, and that empiric anti-HP therapy cannot be justified.

Adolescent↗

[Serological diagnostic trial of the causative virus of Bell's palsy by anti-herpes virus antibodies in the paired sera].

Substantial evidence obtained through polymerase chain reaction techniques strongly supports that reactivation of latently infected herpes simplex virus (HSV) in geniculate ganglia is the main cause of Bell's palsy. However, serum antibody titers to HSV rarely increase in patients with this disease. This discrepancy may result from the difficulty in detecting a small increase in antibody titers by conventional serological analysis. To detect such a small increase in antibody titer, we defined the significant increase in IgG antibody titers more precisely, and examined the association of HSV or varicella-zoster virus (VZV) with Bell's palsy. From 40 patients with Bell's palsy, paired sera were obtained within the 4th disease day and 2 weeks later. IgG antibodies to HSV, VZV, cytomegalovirus (CMV), or measles virus (MsV) were measured with solid-phase enzyme immunoassay (EIA). IgM antibodies were measured with captured EIA. Each antibody titer was expressed as an EIA-value, which was quantitatively measured by using a calibration curve prepared from the samples containing known titers to the virus in proportion to the logarithm of its titer. An EIA-value ratio (EVR) between paired sera and a corrected EVR was calculated according to the formulae: EVR = [EIA-value in the second serum]/[EIA-value in the first serum]; corrected EVR = [EVR to HSV, VZV or CMV]/[EVR to MsV]. The association with a virus was determined to be positive when the corrected EVR of the virus was beyond the normal range (the logarithmical mean +/- 3 SD of corrected EVRs among 24 healthy controls) while the corrected EVR of the other viruses were within it. Using the corrected EVR, 6 patients were positive: 4 for HSV, 2 for VZV. On the other hand, the conventional serological analysis, which confirms positivity by a 4-fold increase in IgG antibody titer or a demonstration of IgM antibody, disclosed only 2 positive patients (1 for HSV, 1 for VZV). EIA is a very sensitive method of detecting antibodies. Corrected EVRs can exclude a decrease in antibody titers induced by corticosteroids, which are generally used for therapy of Bell's palsy. Moreover, the normal range of corrected EVRs can be defined more precisely than in conventional serological analyses. Our results indicate that some sensitive analysis, such as the corrected EVR method, may make it possible to serologically detect the causative virus of Bell's palsy.

Adult↗

The genital mycoplasmas: serological inquiries.

Serological studies on the genital mycoplasmas (U. urealyticum and M. hominis) are briefly reviewed. Newly developed serological tests from our laboratory have been applied to the studies of mycoplasma strains and antibody responses in patients. The data indicate that genital mycoplasmas are serologically diverse, with at least 11 serotypes of U. urealyticum and 7 of M. hominis. No one serotype predominates in relation to any known association with illness. However, serological and cultural data indicate a strong link between genital mycoplasmas and perinatal morbidity and mortality.

Antigen-Antibody Reactions↗

[The clinical significance of negative serological markers of hepatitis B infection in hepatitis B virus carriers with chronic hepatic disease].

OBJECTIVE: To assess the clinical significance of cases who were negative for hepatitis B serological markers but were found incidently to be HBV carriers with HBV replicative infection. METHODS: In 12 out of the 50 biopsy proved cases, the hepatitis B serological markers and anti-HCV were all negative and ALT level also normal in repeated assays. Serum HBV DNA and HCV RNA of all the cases were analysed by PCR. Liver specimens were examined by immunohistochemistory for HBsAg and HBcAg. The pathological changes were classified into 4 grades and 4 stages, and Knodell HAI was also recorded. RESULTS: Five cases initially diagnosed as HBV carriers showed high viral replication (4 cases had loads of 10(4) approximately 10(5) copies/ml, the other 1 had < 10(4) copies/ml). The periportal and/or intralobular inflammation and fibrosis were G1 and S1. The virus load of the 4 cases initially suspected of chronic hepatitis were even higher (2 case > 10(6) copies/ml, 2 case > 10(5) copies/ml). On the contrary, 3 cases initially suspected of liver cirrhosis showed low replication (all less than 10(4) copies/ml), but their histologic changes were more advanced, 2 cases of G4 and S4, 1 case of G3 and S4. Their HAI were 17, 17 and 11, respectively. CONCLUSIONS: HBV carriers with normal level of ALT and negative serological HBV markers are not very uncommon. They often have a high replicative infection. Some of the chronic hepatitis patients with normal ALT and negative serological HBV markers have more active viral replication. The clinical significance of this kind of patients should not be underestimated. It may be the cause of some cases of cryptogenic cirrhosis and hepatocellular carcinoma of unknown etiology.

Adult↗

Treponemal serologic tests; experiences of the Bacteriology Laboratory, California State Department of Public Health.

In a study of the relationship of clinical impression regarding syphilis and age, sex and pregnancy status to treponemal serologic test reactivity, it was noted that in diagnostic "problem cases" the standard lipid serologic test titers did not differentiate between syphilitic and biologic false positive reactors. Preliminary data indicated that heroin addiction may be a source of biologic false reactions and that pregnant women with standard serologic test reactivity have a lower treponemal reactivity rate than other women with lipid serologic reactivity.

Bacteriology↗

[Interpretation of questionable syphilis serologies].

Serologic tests are essential for the diagnosis of syphilis. In symptomatic patients, serodiagnosis confirms the clinical diagnosis. During the asymptomatic stage, interpretation of serologic tests is more difficult, either because of low serologic titres or because of discrepant serologies. Questioning of the patient will help to find a history of syphilis within the previous months or years, an adequate therapy or no treatment at all. Another issue that must be mentioned is the impossibility to differentiate syphilis and non-sexually acquired treponemal infections.

Diagnosis, Differential↗

[Comparative studies of serological typing and HLA-A, B antigen genotyping with PCR using sequence-specific primers].

OBJECTIVE: To evaluate the accuracy of PCR with sequence-specific primers (PCR-SSP) for HLA-I genotyping and analyze the causes of the errors occurring in the genotyping. METHODS: DNA samples and were obtained from 34 clinical patients, and serological typing with monoclonal antibody (mAb) and HLA-A and, B antigen genotyping with PCR-SSP were performed. RESULTS: HLA-A and, B alleles were successfully typed in 34 clinical samples by mAb and PCR-SSP. No false positive or false negative results were found, and the erroneous and missed diagnosis rates were obviously higher in serological detection, being 23.5% for HLA-A and 26.5% for HLA-B. Error or confusion was more likely to occur in the antigens of A2 and A68, A32 and A33, B5, B60 and B61. CONCLUSIONS: DNA typing for HLA-I class (A, B antigens) by PCR-SSP has high resolution, high specificity, and good reproducibility, which is more suitable for clinical application than serological typing. PCR-SSP may accurately detect the alleles that are easily missed or mistaken in serological typing.

Antibodies, Monoclonal↗

Herpes simplex virus serology in an asymptomatic patient.

BACKGROUND: Genital herpes is one of the most common sexually transmitted infections. While genital herpes can present with self limiting genital lesions, most people with genital herpes simplex virus (HSV) infection are asymptomatic or have unrecognised infection. Use of type specific serology for HSV antibodies can identify these individuals. OBJECTIVE: This article discusses the role and use of HSV serology in asymptomatic patients, including pre- and post-test counselling and interpreting results. DISCUSSION: The indications for HSV serology in an asymptomatic patient are limited. Patients tested for HSV antibodies require careful assessment and counselling. Identifying asymptomatic genital herpes should ideally only be done in cases where there may be clinical benefit. Limitations of type specific HSV serology need to be considered when interpreting the results in low risk patients.

Antibodies, Viral↗

Spurious serologic test results in patients receiving infusions of intravenous immune gammaglobulin.

Intravenous immune gammaglobulin (IVIgG) has a definite role in the modern armamentarium of therapies for immunocompromised patients. Intravenous IgG is manufactured from the plasma of a large number of donors and contains antibodies against numerous infectious agents. Patients who receive infusions of IVIgG may test false-positive for a variety of viral, bacterial, and other infectious diseases when serologic determinations are performed on their blood samples. It is important that laboratories performing serologic testing be aware that patients have been infused with IVIgG, so that spurious laboratory results can be identified. Serologic testing on implicated lots of IVIgG and on the patient's pretreatment sample may aid in correctly identifying a spurious serologic result due to therapy with IVIgG.

Adolescent↗

Demonstration of PGL I antigens in skin biopsies in indeterminate leprosy patients: comparison with serological anti PGL I levels.

The survey of histological findings in leprosy patients from 1985 to March 1988 has been carried out at the Pasteur Institute in Noumea. Histologically, according to Ridley Jopling criteria, 82 patients were classified, 14 as T.T., 12 as B.T., 2 as B.B., 7 as B.L., 4 as L.L.s., 24 as L.L.p. and 19 as Indeterminate leprosy. Histological examination of tissues sections, using a monoclonal anti-PGL I antibody showed PGL I antigen in histiocytic cells of the infiltrate and more rarely in Langerhans cells in 7 cases of indeterminate leprosy. The Ziehl staining method revealed the presence of alcoholo resistant bacilli in only one case. For 10 patients, histological findings were compared with serological results. In 3 cases, the diagnosis was confirmed by the 2 techniques (immunohistology and serology). In 2 cases by only immunohistology or by serology and in 3 cases the diagnosis was not confirmed by either methods. These results showed the interest of the immunohistological and serological methods in indeterminate leprosy, specially in children. A study of household contacts may be of interest.

Adolescent↗

Observations on the interpretation of amoebic serology in endemic areas.

Antibody to Entamoeba histolytica antigen was evaluated in cases of proven and suspected intestinal amoebiasis as well as in extra-intestinal cases from an endemic area. Screening methods included the gel diffusion precipitation test (GD), counter immunoelectrophoresis (CIE) and the indirect haemagglutination test (IHA). Control populations consisting of asymptomatic cyst passers, non-amoebic individuals, patients with giardiasis and cases of enteric fever were also screened using the above tests. All (100%) cases of amoebic liver abscess and 75-80% of hepatic amoebiasis without overt abscess formation could be detected by serology, with good correlation between the tests used. However, the interpretation of serology in cases of proven and suspected intestinal amoebiasis posed two main problems. The presence of low antibody levels even in proven cases and the persistence of antibodies due to past infection was found to reduce the diagnostic efficiency of serology for acute intestinal amoebiasis. There was no statistical difference between intestinal cases and controls when comparing either percentage of cases detected or titre of antibody obtained. The results of this study indicate that serology for acute intestinal amoebiasis can be unreliable; however, it is of undoubted value as an adjunct to clinical diagnosis in cases of hepatic amoebiasis.

Animals↗