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[Study of serological and biological markers in viral hepatitis. 157 hemophiliacs].

We observed for a two years period 157 hemophiliacs (138 with hemophilia A whose 13 were severe and 19 with hemophilia B whose 13 were severe) and we studied the incidence of liver dysfunction and the role played by HB and non-A, non-B, viruses. Whereas 32 patients not related had no evidence of serological HB virus markers (by radioimmunoassay), 88 (70,4 %) among the 135 hemophiliacs with large or small exposure to blood products were "HB positive". 90,9 % were positive for anti-HBs and anti-HBc antibodies and only two patients had persistent antigenemia. These results appeared independent of the kind of treatment (factor VIII or factor IX concentrates). Six among 17 children born since 1974, when the antigen was detected by RIA, had the serological HB virus markers, showing that this method is not sufficient to completely eliminate the HB virus. However the amount of viruses injected is too small to induce an acute hepatitis and rather produces specific antibodies which protect hemophiliacs against reinfection. An elevated level of serum transaminases (SGPT) was observed in 9,4 % of non treated hemophiliacs, 15,1 % of treated hemophiliacs with no serological markers of HB virus and 27,7 % of treated hemophiliacs "HB positive". This shows that the use of concentrates and the occurring of HB virus in the patients are not the only factors producing liver dysfunction. The role of non-A, non-B viruses has been recognized in 7 patients out of 9 with transient elevation of serum transaminase levels, by Trepo with an immunodiffusion technique.

Acute Disease↗

Evaluation of six commercially available kits using purified heterophile antigen for the rapid diagnosis of infectious mononucleosis compared with Epstein-Barr virus-specific serology.

BACKGROUND: Novel commercial kits based on antibody reactivity to purified heterophile antigens have recently been introduced for the diagnosis of Epstein-Barr (EB) virus-associated infectious mononucleosis (IM). It is important to determine possible improvements in the performance and reliability of such tests for the diagnosis of IM. OBJECTIVE: To evaluate the reliability of six commercially available kits for the rapid diagnosis of IM in comparison to EB-virus-specific serology. STUDY DESIGN: In total, 100 sera, 53 from patients with serologically verified primary EB virus infection and 47 from EB-virus-immune or -susceptible patients, were used to evaluate the six rapid test kits: Monolatex, Mono-Latex, Mono-Lex (latex agglutination-based kits), Mono-Plus, IM-Check and Clearview IM (solid-phase-based kits). EB-virus-specific serologies including detection of viral capsid antigen IgM and IgG and EB nuclear antigen-1 IgG, were used as reference methods. RESULTS: Compared with the reference methods, the sensitivities and specificities of the heterophile antibody test kits were 70-92% and 96-100%, respectively. IM-Check had a low sensitivity and was difficult to read. The remaining kits performed well. CONCLUSION: Monolatex, Mono-Latex, Mono-Lex, Mono-Plus and Clearview IM can be recommended for the confirmation of EB-virus-associated infectious mononucleosis. Clearview IM combined a high sensitivity and specificity with very simple one-step solid-phase-based procedure.

Adolescent↗

Evaluation of epidemiological and serological predictors of human immunodeficiency virus type-1 (HIV-1) infection among high risk professional blood donors with western blot indeterminate results.

BACKGROUND: Indeterminate pattern of results in Western blot (WBI) for human immunodeficiency virus type-1 (HIV-1) infection may represent early HIV-1 infection or may be non-specific in origin. This issue can be resolved by follow up testing upto at least 6 months resulting in psychological distress as well as in high drop out rates among those undergoing investigation pointing out the need for additional parameters that could help in determining the status of HIV-1 infection at the time of initial testing itself in individuals with WBI pattern. OBJECTIVE: The objectives of the present study were: (i) to determine the frequency of HIV-1 infected individuals in a group of professional donors showing WBI patterns in initial testing on the basis of follow up serological studies; (ii) to find out if any HIV related epidemiological or serological characteristics recorded at the time of initial testing could be considered as predictor for HIV-1 infection in WBI specimens; and (iii) to evaluate two alternative serodiagnostic strategies for HIV-1 infection viz. multiple EIAs based on different antigen preparations/principles and a line immunoassay (LIA) employing recombinant antigens in resolving status of HIV-1 infection in specimens showing WBI results at initial testing. STUDY DESIGN: Professional donors with WBI patterns belonging to EIA reactive and EIA nonreactive groups were subjected to study of epidemiological profile, prevalence of sexually transmitted diseases (STD) markers and follow up serological testing for HIV-1 at 6, 12, 24 and 48 weeks intervals to record any seroconversion. The initial and follow up specimens from the donors with initial WBI results were subjected to two EIAs (one based on dot immunoassay using synthetic HIV-1 antigens and other based on microwell EIA using recombinant HIV-1 proteins) as well as to LIA. RESULTS: Professional donors with initial WBI results, from the EIA reactive group had higher proportion of unmarried individuals (90.3%), with history of heterosexual promiscuity (75%) and visit to STD clinics (36.1%) compared with the WBI donors from the EIA nonreactive group (72.7, 42.4 and 12.1%, respectively, P values < 0.001). Prevalence of antitreponemal antibodies was higher in the former group (16.7%) compared with the later group (1.5%, P value < 0.002). Seroconversion was recorded in 4 (7.3%) out of 55 EIA reactive WBI donors from the EIA reactive group that were characterised by high optical density (OD) values in EIA, 'p24 only' pattern of band in WB and positivity by LIA at the time of initial testing. LIA was found to be more reliable test compared with combination of EIAs to determine status of HIV-1 infection in WBI specimens at the time of initial testing. CONCLUSION: The present study points out that parameters like history of heterosexual promiscuity, prevalence of STD markers, high OD values in screening EIA, 'p24' only pattern of bands in WB and positivity by LIA could have individual predictive values for HIV-1 infection in specimens showing WBI pattern of results at initial testing.

AIDS Serodiagnosis↗

Serological evidence of endemic waterborne cryptosporidium infections.

PURPOSE: Cryptosporidium oocysts are commonly detected in surface-derived drinking water, however, the public health significance of these findings is unclear. This study compared the evidence of prior Cryptosporidium infection for people drinking water derived from surface versus ground water sources. METHODS: This study measured serological responses to two Cryptosporidium antigen groups for blood donors from two midwestern United States cities with different drinking water sources: filtered and chlorinated river water receiving agricultural and domestic sewage upstream versus chlorinated water from an underground aquifer. Initial and nine-month paired serological responses to two Cryptosporidium antigen groups were compared. RESULTS: Initially, donors from the surface water city had a higher relative prevalence (RP) of a serological response (54% vs. 38%, RP = 1.39 (1.21,1.60)). Donors with a detectable baseline response who resided in the surface water city had a higher relative risk (RR) of an increased intensity of response on the follow-up blood draw (15/17-kDa, 40/100 vs. 11/100, RR = 3.78 (1.89,7.58)), (27-kDa 38/100 vs. 18/100, RR = 2.07 (1.31,3.25)). Donors with no baseline response to the 15/17-kDa marker who resided in the surface water city also had a higher risk of serconversion (38/100 vs. 14/100, RR = 2.63 (1.78,3.91)). CONCLUSIONS: These rates of both background and drinking water related Cryptosporidium infections are substantially higher than previously estimated, however, the risk of illness from infection may be lower.

Animals↗

Section 1A: Rh serology. Coordinator's report.

One hundred forty-two Rh-specific monoclonal antibodies (Mabs) were evaluated by serology in 27 laboratories. Evaluators were asked to test each Mab at three dilutions in specified serological techniques against normal positive and normal negative phenotype cells, and any Rh variant cells that they had available. Raw data was submitted to the coordinator for overall analysis. Results were analysed by expressing the sum of reaction grades for each Mab with each variant cell as a percentage of the sum of reaction grades of that Mab with normal phenotype cells. Anti-D Mabs were sorted into 23 groups which had the same pattern of reactions with different partial D phenotype cells. Eighteen of these corresponded to previously defined patterns; five were new patterns. Combined with data from the previous workshop, this means that 30 different reaction patterns have been defined. A new nomenclature is introduced for numbering the epitopes. Reactions with new variants DNB, DNU and DAR indicated some further subsplits of these patterns. Reactions with Category Va cells indicated that there were five different types of Va cells that could be distinguished serologically with monoclonal antibodies. No patterns of reactivity corresponding to the epitope groups could be observed with the different types of weak D tested. Anti-E Mabs were sorted into 14 groups, and the E variant cells into seven groups.

Antibodies, Monoclonal↗

A serological evaluation of 1979-1982 Kenyan foot-and-mouth disease type SAT 2 viruses.

Serological evaluations of foot-and-mouth disease type SAT 2 viruses isolated in Kenya between 1979 and 1982 were performed using the two-dimensional microneutralization test. Nine field isolates of epizootiological significance were compared with four vaccine viruses. The results obtained identified Tan 5/68 as the most appropriate reference vaccine virus strain since it had the broadest serological spectrum. Potent Tan 5/68 vaccines would be expected to provide adequate protection against the contemporary SAT 2 field viruses. In the case of K183/74, which also was shown to have a broad spectrum with viruses isolated in Kenya, the results show that the 1982 isolate from central Kenya was significantly divergent (r less than 1.00 at P = 0.01) and warranted tactical revaccination for its control. The study highlighted the fact that strain R1215 which had been isolated from the oesophageal-pharyngeal swabs of asymptomatic carrier cattle had a narrow serological spectrum suggesting that such viruses could be unsuitable as vaccine for the national campaign.

Animals↗

Biochemical and serological investigations on clinical isolates of klebsiella.

A series of 925 clinical isolates of klebsiella was examined by serological and biochemical typing. To perform serological typing (capsular swelling) 77 capsular antisera were prepared, tested against the type strains and grouped in 13 pools. With this serotyping method 80% of the cultures were typable and 63 distinct types could be recognized. All strains were typable biochemically by means of the numerical coding system of the API-20E system supplemented by digits derived from 15 additional conventional biochemical tests. With the API-20E system 24 different biotypes could be distinguished whereas the combination of API-20E and the 15 additional tests produced 93 biotypes. Maximum discrimination of strains was achieved by the combination of serological and biochemical typing (256 bioserotypes). The reproducibility, typability and discriminating power of the biotyping system was not inferior to serotyping. For epidemiological purposes biotyping can replace serotyping of Klebsiella species, especially in laboratories less well equipped.

Antibodies, Bacterial↗

Epidemiology of canine leishmaniasis: a comparative serological study of dogs and foxes in Amazon Brazil.

The paper describes a longitudinal field study of canine leishmaniasis in sympatric domestic dog and crab-eating fox (Cerdocyon thous) populations. Dogs were studied in house-to-house surveys, and foxes were studied by live-trapping and radio-telemetry. Because serological tests (IFAT in this case) for leishmaniasis are often of uncertain sensitivity and specificity, we draw conclusions comparatively. Both cross-sectional (age-prevalence) and longitudinal analyses indicate that incidence in dogs was highest in the dry season. Seasonal changes in the age-prevalence relationship for dogs suggest that serological conversion and recovery rates decline with prior exposure to infection, where 'recovery' may be due to loss of a positive antibody titre or death from leishmaniasis. The mean incidence in dogs was higher in the rural than in the urban population and higher in hunting dogs than pet dogs. Cross-sectional and longitudinal analyses of fox serological data suggest that foxes remain positive for longer than dogs on average, either because detectable antibody is more persistent or because they experience a lower mortality rate due to leishmaniasis.

Animals↗

The incidence of hepatitis C virus positive serological test results among cornea donors.

Purpose. We have noticed that the incidence of positive serological tests of hepatitis C virus (HCV) among cornea donors in our eye bank is higher than expected. The purpose of this study was to determine the incidence of these positive findings and identify the contributory risk factors.Methods. All corneas procured between June 1993 and June 1997 were included in this retrospective study. In all cases a routine work-up, including serological testing of donors' sera and evaluation of the procured corneas, was performed prior to corneal transplantation. Donors found to be positive for HCV antibodies were compared to our general population of corneal donors, with respect to the demographic, serological and ophthalmological data obtained during the transplantation work-up.Results. All 851 corneas procured from 438 donors were included. Antibodies to HCV were found in 29 donors (6.6%). Following positive test results, 57 corneas (6.7%) were discarded. The time from donors' death to cornea procurement was significantly longer among HCV-positive patients than in the general donor population (12.3 vs. 9.3 h, p<0.0003). No other differences were detected between the two groups.Conclusion. Delay in harvesting of donated corneas may give rise to false positive HCV-antibodies test results, which may be partially responsible for the high rate of positive findings among cornea donors, with consequent tissue wastage.

Journal Article↗

Differences in serological IgA responses to recombinant baculovirus-derived human papillomavirus E2 protein in the natural history of cervical neoplasia.

Infection with certain types of human papillomavirus (HPV) presents a high risk for the subsequent development of cervical intraepithelial neoplasia (CIN) and cervical carcinoma. Immunological mechanisms are likely to play a role in control of cervical HPV lesions. The HPV E2 protein has roles in virus replication and transcription, and loss of E2 functions may be associated with progression of cervical neoplasia. Accordingly, it is of interest to monitor immune responses to the E2 protein, and previous studies have reported associations between serological reactivity to E2 peptide antigens and cervical neoplasia. In order to investigate serological responses to native, full-length E2 protein, we expressed HPV-16 E2 proteins with and without an N-terminal polyhistidine tag using the baculovirus system. Purified HPV-16 E2 protein was used to develop enzyme-linked immunosorbent assays to detect serological IgG and IgA responses in cervical neoplasia patients and controls. We found that serum IgA levels against the E2 protein were elevated in CIN patients relative to normal control subjects but were not elevated in cervical cancer patients. Moreover, there appeared to be a gradient of response within cervical neoplasia such that the highest antibody levels were seen in lower grades of neoplasia up to CIN 2, whereas lower levels were observed in CIN 3 and still lower levels in cervical carcinoma. These findings suggest that the IgA antibody response to E2 may associate with stage and progression in cervical neoplasia.

Adult↗

Serological response to Helicobacter pylori in gastric and non-gastric cancer.

1. We aimed to evaluate the seroprevalence of Helicobacter pylori (H. pylori) in gastric cancer, non-gastric cancer and outpatients by ELISA and iso-electric focusing, and to compare histology and serology for H. pylori in gastric cancer and outpatients. 2. In 124 patients with gastric cancer, 78 patients with non-gastric cancer and 110 outpatients, H. pylori seroprevalence was assessed by ELISA and isoelectric focusing. Gastric cancer and outpatients underwent endoscopy with biopsies. 3. Seroprevalence by ELISA was significantly higher in gastric cancer compared with non-gastric cancer (84% versus 56%, P < 0.001) but not with outpatients (84% versus 74%). Iso-electric focusing detection of H. pylori was comparable to ELISA: 85, 51 and 75% in gastric cancer, non-gastric cancer and outpatients respectively. Oligoclonal iso-electric focusing was significantly more frequent in gastric cancer compared with non-gastric cancer and outpatients: 69% versus 45 and 46% respectively, P < 0.01. The reliability of H. pylori detection by antral biopsy was significantly lower in gastric cancer compared with outpatients: 36% versus 74% (P < 0.001). In gastric cancer, ELISA and iso-electric focusing were significantly more reliable than histology in H. pylori detection (84 and 85% versus 36% respectively) (P < 0.001). 4. Serological immune response to H. pylori in gastric cancer, non-gastric cancer and outpatients seems different both quantitatively and qualitatively; serology was more reliable than histology in detection of H. pylori in gastric cancer.

Adult↗

Variable number of pig MHC class I genes in different serologically defined haplotypes identified by a 3'-untranslated region probe.

To estimate the number of porcine class I major histocompatibility genes, a short class I cDNA probe from the 3'-untranslated region was developed to be used in restriction fragment length polymorphism analysis. Six clones isolated from a pig spleen cDNA library were sequenced from their 3'-untranslated region. Three different transcripts were identified, one probably derived from the class I PD7 locus and two showing highest homology to the PD1 and the PD14 genes, respectively. Class I typing was performed both by restriction fragment length polymorphism and serology. Segregation of class I haplotypes was followed in one three-generation family (European Wild Boar x Large White: Swedish Yorkshire) and in six two-generation families (Duroc, Yorkshire and Chester White), for a total of 266 pigs. Twenty different class I haplotypes were identified either with restriction fragment length polymorphism and/or serological typing. Furthermore, previously unpublished serological haplotypes H62, H67 and H68 were identified. Two to seven polymorphic and three monomorphic fragments were detected in different restriction fragment length polymorphism haplotypes indicating that the number of class I genes in the investigated haplotypes varies.

Animals↗

Serological diagnosis of Trypanosoma rangeli infected patients. A comparison of different methods and its implications for the diagnosis of Chagas' disease.

Venous blood from 65 Panamanian schoolchildren living in an area endemic for both Trypanosoma cruzi and T. rangeli were screened for the presence of these parasites. Trypanosoma rangeli were isolated and cultured from four individuals. Serological tests of all 65 sera were performed, including immunohaemagglutination (IHA), indirect immunofluorescence assay (IF) and ELISA using both T. rangeli and T. cruzi as antigens, as well as T. cruzi synthetic peptides in an ELISA assay. Results indicated a higher immunoreactivity to T. rangeli preparations than to T. cruzi within the studied population, which could be divided into four 'serological responder' groups. Interestingly, the panel of SAPA and other T. cruzi synthetic peptides were not useful in the discrimination of patients. Furthermore, patients from whom parasites had been isolated could not be distinguished from those of two other groups. Significant immunoreactivity to T. cruzi preparations was displayed in all responder sera, despite total lack of evidence of infection with this parasite. The immunobiological significance of T. rangeli infection is unclear, but these data indicate that it is a compounding problem in the accurate diagnosis of pathological T. cruzi infection by serological analysis. The relationship of these cohabiting species, in respect to infection outcome and immunological activation, is discussed.

Amino Acid Sequence↗

Vaccine-modified measles in previously immunized children in Accra, Ghana: clinical, virological and serological parameters.

Despite rapidly increasing measles immunization coverage, epidemics of measles occurred from January to March 2000 in some parts of Accra, the capital of Ghana. 44 cases of acute measles were diagnosed at three health facilities during the outbreaks, which we examined clinically and serologically. The peak incidence occurred among 6-12-year-olds, clinical symptoms were milder than the typical symptoms of measles, and fever was significantly less common. None of the cases developed complications and all recovered completely. Thirty-eight (86.4%) were tested serologically; IgM antibodies were detected in 73.7% and IgG antibodies in 84.2% during the acute phase. Milder symptoms in a significant number of cases with measles IgG antibodies suggest that these are vaccine-modified measles, attributable to waning antibodies and low circulation of wild type virus in an area of high vaccine coverage. Serological confirmation will be required for accurate diagnosis, if measles is to be eradicated or kept under control. It also seems likely that multiple dose immunization schedules will be needed in the future to maintain protective antibody levels and to protect children against measles in Ghana. This will eliminate the frequent outbreaks of measles involving immunized children.

Child↗

Structural and serological studies on a new 4-deoxy-d-arabino-hexose-containing O-specific polysaccharide from the lipopolysaccharide of Citrobacter braakii PCM 1531 (serogroup O6).

The O-specific polysaccharide of Citrobacter braakii PCM 1531 (serogroup O6) was isolated by mild acid hydrolysis of the lipopolysaccharide (LPS) and found to contain d-fucose, l-rhamnose, 4-deoxy-d-arabino-hexose and O-acetyl groups in molar ratios 2 : 1 : 1 : 1. On the basis of methylation analysis and 1H and 13C NMR spectroscopy data, the structure of the branched tetrasaccharide repeating unit of the O-specific polysaccharide was established. Using various serological assays, it was demonstrated that the LPS of strain PCM 1531 is not related serologically to other known 4-deoxy-d-arabino-hexose-containing LPS from Citrobacter PCM 1487 (serogroup O5) or C. youngae PCM 1488 (serogroup O36). Two other strains of Citrobacter, PCM 1504 and PCM 1505, which, together with strain PCM 1531, have been classified in serogroup O6, were shown to be serologically distinct from strain PCM 1531 and should be reclassified into another serogroup.

Animals↗

Neospora caninum infection and congenital transmission: serological and parasitological study of cows up to the fourth gestation.

In this paper, a parasitological and serological study performed in three cows up to the fourth gestation is reported in order to clarify the extent of vertical propagation and, secondly, in which period of gestation the recrudescence of previous infection occurs. The cows selected for the study delivered healthy but congenitally infected calves in first pregnancy. The parasite was found, by biological tests in Swiss mice, in all the placentas of the three cows examined, during the three subsequent gestations, at calving. The parasite was found, at slaughtering, in the brains of all nine calves born clinically healthy from the three cows as well. The serological profile, performed at monthly intervals on serum of the cows, showed that IgG and IgM increased in the third trimester of gestation; this rise of antibodies was constantly observed during the three gestations and in all three cows. In the calves, the IgG titres increase after colostrum consumption and an IgM peak at birth, were indicative of a late infection. These findings, along with negative results obtained by a serological study conducted simultaneously on 38 cows housed in the same stable as the experimental animals and the negative results obtained in isolating parasite or antibodies from farm dogs, suggest that N. caninum infection can be maintained over several bovine generations and that recrudescing persistent infection, rather than a new infection, explains the Neospora infection of calves.

Animals↗

Epitope specificities and quantitative and serologic aspects of monoclonal complement (C3c and C3d) antibodies.

Twenty-two monoclonal antibodies to human C3c and ten to C3d were obtained by hybridization after the immunization of mice with complement-coated human red cells and/or purified human complement components. C3c antibodies were variable in their agglutination reactions with cells coated with C3 by antibody in vitro; more consistent and potent reactions with these cells were observed with anti-C3d, and all anti-C3d reacted with red cells coated with C3 in vivo. Immunoradiometric assays were used to estimate antibody concentration, affinity, and epitope specificity. The antibody content in ascitic fluids varied from less than 0.1 mg per ml to 5.6 mg per ml. The estimated values of antibody affinities for Sepharose-coupled C3 ranged from 2.8 X 10(6) l per M to 5.0 X 10(8) l per M; on average, IgM antibodies had higher affinities than IgG antibodies. Competitive binding assays showed that the monoclonal antibodies recognized at least seven different epitopes, four on the C3c and three on the C3d fragment of C3. When the results of serologic and quantitative assays were compared, no convincing relationship was found between serologic performance and epitope specificity, antibody concentration, or affinity. IgM antibodies generally gave higher agglutination scores than IgG antibodies, and Ig class was the only useful predictor of serologic efficacy.

Animals↗

Transmission of serologically silent hepatitis B virus along with hepatitis C virus in two cases of posttransfusion hepatitis.

The polymerase chain reaction (PCR) was used to investigate the presence of hepatitis B virus (HBV)-related DNA sequences in blood from three blood donors and two transfusion recipients who developed posttransfusion non-A, non-B hepatitis (NANBH). In the first case, the sole donor was positive for antibody to hepatitis B surface (HBs) and core (HBc) antigens and had elevated alanine aminotransferase (ALT) levels, while the recipient had no HBV serologic markers. Both the donor and the recipient had serologic markers of hepatitis C virus (HCV) and were found positive for HBV DNA and HCV RNA sequences by PCR. The second case involved two donors and one recipient. Serologic tests for conventional HBV markers were negative in all three individuals, but one of the donors had elevated ALT. HBV DNA sequences were detected by PCR in the serum of the recipient and of the donor with high ALT, but not in the serum of the donor with normal ALT. Anti-HCV was detected in the serum of the recipient and of the suspect donor but not in that of the donor with normal ALT. The sequences amplified in the S region and determined after cloning of PCR products for both donor-recipient pairs were indistinguishable from each other and identical to the sequence of the major HBV subtype of adw in the first case and ayw in the second case. Furthermore, for the second case, an identical single-point mutation was found in both the donor and the recipient. These data confirm the transmission of conserved HBV sequences together with HCV in posttransfusion NANBH.(ABSTRACT TRUNCATED AT 250 WORDS)

Alanine Transaminase↗