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PubMed1999

Molecular, serological and population studies of the alleles and products of HLA-B*41.

The HLA-B41 specificity was first identified over 25 years ago and, although both serological and biochemical studies have suggested its subdivision, it is only recently that two HLA-B*41 alleles (B*4101 and B*4102) have been identified and sequenced. We designed three oligonucleotide primers, combined in two mixtures to define these alleles by PCR using sequence-specific primers (PCR-SSP) in a random normal population of 9,464 HLA-A, B, DR, DQ typed Northern European Caucasoid donors from the Welsh Bone Marrow Donor Registry. The HLA-B41 phenotype frequency was 0.835%, and of the 79 HLA-B41 subjects 22 (27.85%) were B*4101 and 57 (72.15%) were B*4102. The phenotype frequencies of B*4101 and B*4102 were 0.232 and 0.602%, respectively, and the gene frequencies were 0.00116 and 0.00301, respectively. Formal two-locus linkage disequilibrium (LD) analysis demonstrated significant associations between B*4101 and A30 and DR11, and between B*4102 and A66 and DR13. LD analysis of three loci showed significant associations between B*4101, DR7, DQ2 and B*4101, DR11, DQ7 (DQB1*0301/0304) and between HLA-A3, B*4102, DR13; A66, B*4102, DR7; A66, B*4102, DR13; B*4102, DR13, DQ6 and B*4102, DR13, DQ7. Examination of the HLA phenotypes (including HLA-C) of the B*41 subjects, together with the LD analysis findings, suggested four different HLA haplotypes bearing B*4101 and five B*4102 haplotypes. The most frequent B*4101 haplotype was: HLA- A30 or other A allele, Cw*1701, B*4101, DRB1*1102, DQB1*0301 and the most freqent B*4102 haplotype was: A*6601 or A3 or other A allele, Cw*1701, B*4102, DRB1*1303, DQB1*0301. In addition, the well-known association of A66 with B41 was between A*6601 and B*4102, and although both B*41 alleles were commonly in association with Cw*1701, B*4101 was found with Cw*07. One-dimensional isoelectric focusing (1D-IEF) analysis of HLA-B proteins from 2 B*4101 and 2 B*4102 subjects clearly showed that the B41.1 and B41.2 1D-IEF variants, identified in the 10th International Histocompatibility Workshop, corresponded to B*4101 and B*4102 products, respectively. Serological titration studies, with 59 lymphocytotoxic pregnancy antisera, containing an HLA-B41 component and stimulated by up to five different HLA-B specificities, were unable to differentiate the two groups of B*41 subjects. This suggests that partition of the HLA-B41 specificity will not normally be achieved by classical serological methods. It is suggested that the previous alleged serological subdivision of HLA-B41 was founded on the unwitting use of antisera detecting the HLA-Cw*17 products.

Alleles↗
PubMed1992

Serological identification of HLA-B13 subtypes.

A serological approach is used to confirm subtypes of HLA-B13 originally observed by one-dimensional isoelectric focusing (1D-IEF). Sixty anti-B13 alloantibody sera were screened against Chinese panel cells. Two clusters of sera showing distinct reactive patterns were identified. One is a shorter reactive pattern than the other. Using these serological reaction patterns, the B13 antigen can be divided into two subtypes, B13.1 and B13.2, in the Chinese population. These serological subtypes appear to correlate well with the 1D-IEF patterns of B13 subtypes. The serological subtyping is also in agreement with the differences in nucleotide sequence previously determined to exist in B13 antigen subtypes. Family studies show that both B13.1 and B13.2 segregate as HLA-B locus alleles. Gene frequencies for B13.1 and B13.2 were 0.0676 and 0.0612, respectively, in our study population of 337 southern Han Chinese.

Alleles↗
PubMed1991

Serological profiles as prognostic clues for progressive systemic scleroderma: the Italian experience.

Ninety-one patients with progressive systemic sclerosis have been examined both clinically and serologically in order to have a better prognostic insight. Three main serological profiles have been isolated. The patients with anticentromere antibodies (ACA) represented one third of the cases, developed skin sclerosis rather later and rarely exhibited ankyloses and ulcerations. The esophagus was commonly involved while the lung, heart and kidneys were not. ACA-positive patients were not identified with the CREST syndrome, as the latter disclosed other profiles with the same frequency. Patients with anti-Scl-70 antibody represented one fourth of the cases and had the fastest progression, developing sclerosis in less than 5 years after the onset of Raynaud's phenomenon. Ankyloses and lung fibrosis, as well as joint, heart and kidney involvement, were found in most of them. Patients with anti-SSA/Ro antibodies were uncommon, but corresponded to a severe subset, having a fast progression and a constant involvement of the lung. Probably due to the rougher definition of their serology, patients with antinuclear, antispeckle-patterned and anti-Ku antibodies or without any detectable antibody could be defined less easily and corresponded to an intermediate position between ACA- and anti-Scl-70-positive patients. Though it is probably premature to trust it completely, a serological classification may provide the prognostic clues clinical classifications cannot.

Adult↗
PubMed1990

Significance of false-positive serologic tests for histoplasmosis and blastomycosis in an endemic area.

False-positive serologic tests for histoplasmosis (H) and blastomycosis (B) are common in populations from endemic areas. In order to determine the significance of false-positive test results, we reviewed the final diagnoses of all patients whose sera were submitted to our laboratory for radioimmunoassay (RIA) and immunodiffusion (ID) during a 3-yr period. Of the 263 patients whose sera were examined, 29 (11%) had H or B; 41 (17.5%) of the remaining 234 patients had false-positive test results. Of these 41 patients, 31 were positive for H alone, and 10 had antibodies to both H and B. All three patients with false-positive ID tests for histoplasmosis also had positive titers (greater than or equal to 1:16) on RIA. No patient had a false-positive ID result for blastomycosis. The percentage of patients in each of five major diagnostic categories with and without false-positive serologic tests was similar (p greater than 0.05). The majority of patients had pulmonary infections, almost half of which were granulomatous infections other than H or B; this reflects the clinical indications for requesting fungal serologic tests. A positive fungal serology is not useful in suggesting the presence of a pulmonary disease other than H or B in patients from an endemic area suspected of having a pulmonary mycosis.

Blastomycosis↗
PubMed1995

The role of serology in the diagnosis of Helicobacter pylori infection in children.

The aim of this study was to evaluate the diagnostic accuracy of serologic tests and to verify whether specific IgG antibody can be used in the diagnosis of Helicobacter pylori infection in children without endoscopy. Serologic and endoscopic examinations were performed in 310 children (1 month-15 years) with gastrointestinal symptoms. Serologic outcomes were compared with the results of rapid urease test, histologic examination, and culture. The prevalence of H. pylori infection was 21.0% and the seropositivity of H. pylori was 24.2% in these symptomatic children. Serologic examination for diagnosis of H. pylori showed a sensitivity of 94.9% and a specificity of 92.4%, diagnostic reliability equal to the rapid urease test and histologic examination in children. The serum IgG test is a useful tool for the diagnosis of H. pylori infection, especially in children.

Adolescent↗
PubMed2001

Detection of chlamydial antibody by fetal serology--an aid to the diagnosis of ovine abortion.

Two serological tests (indirect immunofluorescence and enzyme-linked immunosorbent assay) were developed for the detection of fetal antibody to Chlamydia psittaci. Fetal blood and thoracic fluid from 126 field cases of suspected ovine chlamydial abortion were examined using both tests. Placenta and fetal tissues (lung, liver, and kidney) from the same animals were also examined by the following conventional diagnostic methods: isolation in McCoy cells, detection of chlamydial lipopolysaccharide (LPS), modified Ziehl-Nielsen staining, and direct fluorescent antibody staining of chlamydia in frozen cryostat sections. Seventy cases were positive by fetal serology, and of these, 68 were also positive by isolation and/or LPS detection. The remaining 56 cases had negative fetal serology, and of these, 39 were positive by isolation and/or LPS detection. Results indicate that fetal serology, although less sensitive than either isolation in McCoy cells or detection of chlamydial LPS antigen, may be of particular use when placenta is not available.

Abortion, Veterinary↗
PubMed2001

Isolation and serological identification of enteropathogenic Escherichia coli in pasteurized milk in Brazil.

OBJECTIVE: To evaluate the microbiological quality of pasteurized milk commercialized in Rio de Janeiro, Brazil, and determine serologically enteropathogenic Escherichia coli (EPEC) strains in E. coli isolates obtained from milk samples. METHODS: Ninety samples of pasteurized milk - types B and C - of three different commercial brands, purchased in supermarkets and bakeries in Rio de Janeiro, were examined. The amount of total and fecal coliform bacteria was estimated using the Most Probable Number technique. Mesophilic, psychrotrophic, and thermoduric microorganism counts were determined by the Standard Plate Count technique. Isolation and identification of E. coli were carried out using conventional physiological tests. Commercial antisera were used for serological characterization of EPEC. RESULTS: The three milk brands analyzed revealed bacterial counts above the regulated values of the Brazilian government. It was found that among 208 strains of E. coli isolated, 46 (22.1%) were serologically classified as EPEC. The most common EPEC serogroup was O55 (15.2%). CONCLUSIONS: Though recent studies on virulence factors indicate that not all strains serologically classified as EPEC are able to attaching/effacing lesion, it is believed that the isolation of EPEC serogroups from pasteurized milk represent a potential risk for children, as well as an indicative of the presence of other enteropathogens.

Animals↗
PubMed1995

Uses of serology for the diagnosis of contagious bovine pleuropneumonia.

A serological test involves detection of specific changes, induced by a pathogen, in the properties or actions of serum of an infected host. The test may detect the presence in serum of either antibodies to the pathogen (produced by the host) or antigens (i.e. the infecting agent itself and/or its components). The many serological tests which have been developed for the diagnosis of contagious bovine pleuropneumonia (CBPP) are classified into two groups on the basis of this distinction. To date, no single serological test is able to detect all stages of the disease. Thus the choice of serological test (or combination of tests) will depend on the specific aim of the investigation. Meanwhile, a sensitive, specific and simple 'pen-side' test for the diagnosis of all forms of CBPP is still lacking.

Animals↗
PubMed2002

Serologic evaluation of five unvaccinated heifers to detect herds that have cattle persistently infected with bovine viral diarrhea virus.

OBJECTIVE: To determine whether serologic evaluation of 5 unvaccinated 6- to 12-month-old heifers is a valid method for identifying herds that contain cattle persistently infected (PI) with bovine viral diarrhea virus (BVDV). ANIMALS: 14 dairy herds with a history of BVDV infection, with health problems consistent with BVDV infection, or at risk for contracting BVDV infection. PROCEDURE: 5 unvaccinated 6- to 12-month-old heifers were randomly selected from each herd. Neutralizing antibody titers for type-I and -II BVDV were determined. A herd was classified as likely to contain PI cattle when at least 3/5 heifers had antibody titers > or = 128. Virus isolation was performed on all cattle to identify PI cattle. Genotype of isolated viruses was determined by nested multiplex polymerase chain reaction. RESULTS: 6 of 14 herds contained PI cattle. Sensitivity and specificity of serologic evaluation of 5 heifers for identifying these herds were 66 and 100%, respectively. In herds that contained PI cattle, the predominant BVDV titer in the tested heifers corresponded to the genotype of the isolated virus. CONCLUSIONS AND CLINICAL RELEVANCE: Serologic evaluation of unvaccinated 6- to 12- month-old heifers is an accurate method for identifying herds containing PI cattle. Both type-I and -II BVDV antibody titers should be determined to prevent herd misclassification. The genotype of BVDV found in PI cattle can be predicted by the predominant neutralizing antibody titers found in tested heifers. Serologic evaluation of 5 unvaccinated heifers can be used to determine whether a herd is likely to contain PI cattle.

Animal Husbandry↗
PubMed1981

Chlamydial isolations and serology in Reiter's syndrome.

To assess the possible etiological link between genital infection due to Chlamydia trachomatis (CT) and Reiter's syndrome (RS) 24 men and 2 women with typical RS and 5 women with signs suggestive of RS (SRS) were examined. CT was isolated by irradiated McCoy cell culture technique and chlamydial indirect immunofluorescence serology was applied. Chlamydial isolation from the urethra was positive in 5 patients as well as from synovial fluid of the knee in one man. The serology was positive (titre greater than or equal to 64) in 17 (55%) of the 31 patients, including all 7 women. HLA-B27 antigen was detected in only 15 of 27 patients (55%). All except one of the 12 patients negative for HLA-B27 antigen had positive chlamydial serology. Among the patients were two married couples, both negative for B27 antigen and positive for chlamydial serology. The case report of the one couple, in which the wife had positive urethral chlamydial isolation, is given. Sexually acquired RS might easily be overlooked in women and be misdiagnosed as seronegative arthritis. The classification for RS associated with CT infection is discussed. The simultaneous treatment of the infection in sexual partners is emphasized.

Adolescent↗
PubMed2003

Trypanosoma cruzi in persons without serologic evidence of disease, Argentina.

Current diagnosis of chronic Chagas disease relies on serologic detection of specific immunoglobulin G against Trypanosoma cruzi. However, the presence of parasites detected by polymerase chain reaction (PCR) in patients without positive conventional serologic testing has been observed. We determined the prevalence and clinical characteristics of persons with seronegative results and T. cruzi DNA detected by PCR in a population at high risk for chronic American trypanosomiasis. We studied a total of 194 persons from two different populations: 110 patients were recruited from an urban cardiology clinic, and 84 persons were citizens from a highly disease-endemic area. Eighty (41%) of persons had negative serologic findings; 12 (15%) had a positive PCR. Three patients with negative serologic findings and positive PCR results had clinical signs and symptoms that suggested Chagas cardiomyopathy. This finding challenges the current recommendations for Chagas disease diagnosis, therapy, and blood transfusion policies.

Adolescent↗
PubMed1994

Reverse-SSO hybridization provides an accurate and simple HLA-DR typing--a comparative study with HLA-DR serologic typing.

The HLA-DR molecule is a polymorphic membrane glycoprotein and one of the key molecules causing allograft rejection and graft-versus-host disease in organ transplantation. Serologic typing using DR specific alloantisera has long been used, but several problems have limited its application. The purpose of this study was to establish an efficient reverse-SSO typing system that detects DRB1 and DRB3/B4/B5 alleles on a single membrane. A DR typing membrane was prepared by immobilizing 21 dT-tailed sequence specific oligonucleotides (SSOs) on a nylon membrane and was used in a hybridization assay with digoxigenin-labeled PCR-amplified target DNA. The positive signals were detected on X-ray film using chemiluminescence. A comparison study with serology using DNAs from 105 unrelated individuals demonstrated that the reverse-SSO typing system was superior to serologic typing in terms of accuracy (100% vs 90.5%), simplicity, range of application, rapidity, and cost of the test. These data indicate that the reverse-SSO typing system can replace serology as a routine DR test, and will be useful in time-restricted solid organ transplantation and in selection of an unrelated marrow donor prior to bone marrow transplantation.

Base Sequence↗
PubMed2006

[Identification of infectious agents in cases suspicious of Hantavirus infection but with negative serology].

BACKGROUND: The diagnosis of acute respiratory illness caused by Hantavirus is based in the rapid and progressive clinical course, epidemiological background and the serological confirmation of the virus. When the presence of the virus is not confirmed a differential diagnosis must be made with other infections. Between 1999 and 2001, the Chilean Public Health Institute received 1063 blood samples from patients with a suspicious clinical picture, to study the presence of Andes strain of Hantavirus. In 134 of these samples, the presence of the virus was confirmed. AIM: To study the presence of other infections in sera from patients with suspected Hantavirus acute respiratory illness but serologically negative for Hantavirus. MATERIAL AND METHODS: A retrospective study of 98 serum samples, received at the National Public Health Institute, of patients with negative serology for Hantavirus. The presence of antibodies against influenza virus, Mycoplasma pneumoniae, Leptospira and Streptococcus pneumoniae was determined using Latex techniques. RESULTS: Leptospira was detected in 23 patients, influenza virus in 13, Streptococcus pneumoniae in six and Mycoplasma pneumoniae in one case. Leptospira infections were confirmed in four cases by ELISA determination of IgM antibodies. Influenza virus infection was confirmed in three cases by Hemmaglutation Inhibition Assay. CONCLUSIONS: In suspected cases of Hantavirus acute respiratory infection, but with negative serology, other infectious agents such as Leptospira, influenza virus and Streptococcus pneumoniae, must be sought.

Adolescent↗
PubMed1995

Comparison of the polymerase chain reaction and serology for the detection of canine visceral leishmaniasis.

The polymerase chain reaction (PCR) and serology was evaluated for the diagnosis of canine visceral leishmaniasis in Bahia, Brazil in a study of 125 dogs. The PCR was 100% sensitive in 25 dogs that had Leishmania demonstrated by either culture or hamster inoculation. It was 100% specific for 35 dogs from the northeastern United States, all were PCR negative. However, 22 of 54 Brazilian dogs that were culture-hamster inoculation-negative were positive by PCR. The nature of the PCR product was identified by hybridization with specific Leishmania probes. Whereas the sensitivity of serology in relationship to infection, as determined by hamster or culture assay was more than 80%, sensitivity of serology was only 63% when compared with PCR. These results raise questions about the use of serology to detect Leishmania infection in dogs, and suggest that the PCR might serve as a better gold standard to define Leishmania infection than culture or hamster inoculation.

Animals↗
PubMed1979

Evaluation of three serological tests for the diagnosis of hydatid disease.

A number of techniques is used for the serological diagnosis of hydatid disease; however previous investigations suggest variable degrees or reliability for each test. This study reports an evaluation of the sensitivity and specificity of three commonly used serological tests for hydatid disease; complement fixation (CF), haemagglutination (HA) and the fluorescent antibody (FA) techniques. The results of these serological tests are compared in a series of specimens from 116 hospital patients, 21 of whom were confirmed surgically as having hydatid disease. A similar high degree of sensitivity was found for each of the three serological tests examined. The CF and HA tests gave negative results in two patients with confirmed hydatid disease, and the result of the FA test was negative in only one patient. The HA test was the most specific with no false positive reactions, but only two false positive reactions occurred with the FA test. The CF test was the least specific as the reaction was positive in nine patients without hydatid disease. When the results of both FA and HA tests were considered together, all 21 cases of hydatid disease were detected and false positive reactions occurred in only two patients.

Complement Fixation Tests↗
PubMed1991

Serologic response to treatment of infectious syphilis.

OBJECTIVE: To evaluate the serologic response to treatment of patients with infectious syphilis. DESIGN: Historical cohort study of all cases of infectious syphilis in Alberta from 1981 to 1987. PATIENTS: A total of 1090 patients were entered; 857 with primary syphilis, 183 with secondary syphilis, and 50 with early latent disease. Two hundred and eight patients were excluded who either were pregnant, had negative serologic results before treatment, had clinical relapse, were treatment failures, or were lost to follow-up. INTERVENTIONS: All 882 evaluable patients were treated with a recommended antibiotic regimen for infectious syphilis and returned for re-assessment including repeat serologic testing. MEASUREMENTS AND MAIN RESULTS: Seventy-two percent (95% CI, 66% to 77%) and 56% (CI, 43% to 70%) of patients with initial episodes of primary or secondary syphilis had seroreverted according to rapid plasma reagin (RPR) test results by 36 months. A 2- and 3-tube decline was seen by 6 and 12 months in primary and secondary syphilis. Early latent syphilis resulted in only a 2-tube decrease at 12 months. Serologic response was not affected by sex, age, race, or sexual orientation. Patients with their first infection were more likely to experience RPR seroreversal than those with repeat infections. The RPR reversal rates also depended on the pretreatment titer and stage of disease. At 36 months, 24% (CI, 20% to 28%) of patients had nonreactive fluorescent treponemal antibody absorption tests (FTA-Abs), and 13% (CI, 11% to 15%) had nonreactive microhemoglutination tests for Treponema pallidum (MHA-TP). CONCLUSIONS: Adequate therapeutic response for syphilis must be based on illness episode and the pretreatment RPR titer. Treponemal tests can demonstrate seroreversion after 36 months, and a negative treponemal test does not rule out a past history of syphilis.

Alberta↗
PubMed1993

Effects of HIV infection on the serologic manifestations and response to treatment of syphilis in intravenous drug users.

OBJECTIVE: To describe the effects of human immunodeficiency virus (HIV) infection on the serologic manifestations and response to treatment of syphilis in intravenous drug users. DESIGN: Cohort study of intravenous drug users. SETTING: Medical clinic in a hospital-based methadone maintenance treatment program in New York City. PATIENTS: Fifty patients with syphilis, of whom 31 were HIV seropositive and 19 HIV seronegative. MEASUREMENTS: Serologic tests for syphilis and clinical manifestations. RESULTS: Stage of syphilis at presentation was not associated with HIV serologic status. No unusual or fulminant manifestations of early syphilis or neurosyphilis were noted among HIV-seropositive cases. Maximum nontreponemal titers were higher among HIV-seropositive (median, 1:128) than among HIV-seronegative (median, 1:32) patients with syphilis (P = 0.05); this difference was present only among patients with first-episode syphilis. All 26 evaluable, HIV-seropositive patients treated for syphilis responded appropriately, including 13 patients given standard or less-than-standard doses of penicillin. Seven of 43 patients (16%) showed reversion to negative treponemal antibody assay results after treatment for syphilis; this finding was not associated with HIV infection, CD4 count, or stage of syphilis. Low nontreponemal titer was weakly associated with treponemal test reversion. CONCLUSIONS: Infection with HIV did not alter the stage at presentation, clinical course, serologic manifestations, or response to treatment of syphilis in this cohort of intravenous drug users.

Adult↗
PubMed1998

Probable false positive coccidioidal serologic results in patients with cystic fibrosis.

Coccidioidomycosis is an acquired fungal infection that afflicts primarily the respiratory tract. Cystic fibrosis patients who are being treated with glucocorticoids and immunosuppressed organ recipients may be at risk for infection with Coccidiodes immitis or reactivation of latent infection. The diagnosis is best made by demonstration of the organism in pathologic specimens or by culture. Serologic screening is another method that is reliable in most patients. We studied 98 patients who had serologic screening for Coccidiodes immitis performed as part of their evaluation for lung transplantation. This study revealed that approximately 15% of the cystic fibrosis patients screened had putative coccidioidal IgM, in the absence of an IgG response. None of the patients studied had a positive fungal culture for the organism. None of the non-cystic fibrosis patients screened had detectable coccidioidal IgG or IgM. We hypothesize that cystic fibrosis patients may have hyperimmune sera which interferes with serologic screening tests. We would recommend repeat serologic testing and attempts to identify the organism in tissue or by culture to confirm the diagnosis in these patients.

Adolescent↗