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The HLA dictionary 1999: a summary of HLA-A, -B, -C, -DRB1/3/4/5, -DQB1 alleles and their association with serologically defined HLA-A, -B, -C, -DR and -DQ antigens.

This report presents serologic equivalents of 90 HLA-A, 190 HLA-B and 145 HLA-DRB1 alleles. The equivalents cover over 70% of the presently identified HLA-A, -B and -DRB1 alleles. The dictionary is an update of the one published in 1997 and now also includes equivalents for HLA-C, DRB3, DRB4, DRB5 and DQB1 alleles. The data summarize information obtained by the WHO HLA Nomenclature Committee, the International Cell Exchange (UCLA), the National Marrow Donor Program (NMDP) and by individual laboratories. In addition, a listing is provided of alleles which are expressed as antigens with serologic reaction patterns that differ from the well-established HLA specificities and that often lack official WHO nomenclature. The provided equivalents will be useful in guiding searches for unrelated donors in which patients and/or potential donors are typed by either serology or DNA-based methods. These equivalents will also serve typing and matching procedures for organ transplant programs where HLA typings from donors and from recipients on waiting lists represent mixtures of serologic and molecular typings. Some guidelines are provided for the use of appropriate WHO HLA nomenclature for serological typings and for generic and allele specific typings obtained with molecular methods. The tables with HLA equivalents and the questionnaire for submission of serology on poorly identified alleles will also be available at the WMDA web page: www:bmdw.org/wmda.

Alleles↗

Validation of DNA-based HLA-A and HLA-B testing of volunteers for a bone marrow registry through parallel testing with serology.

A total of 42,160 individuals were typed for HLA-A and HLA-B by both serology and PCR-based typing. The HLA assignments included all of the known serological equivalents. The majority of the individuals (99.9%) were from U.S. minority population groups. The serologic typing was performed between 1993 and 1997 at the time of recruitment for the National Bone Marrow Program (NMDP) registry. The polymerase chain reaction (PCR)-based typing was carried out in two phases. In phase I, DNA typing was performed by PCR using sequence-specific oligonucleotide probes (PCR-SSOP) or PCR using sequence-specific primers (PCR-SSP) without knowledge of the serologic assignments. Discrepancies were identified between the serologic and DNA assignments in 24% of the volunteers (8% of volunteers differed for only HLA-A assignments, 13% for HLA-B, and 3% for both HLA-A and -B) and a potential explanation was assigned each discrepant serology/DNA pair. In phase II, a random sampling scheme was used to select a statistically significant number of individuals for repeat DNA typing from each of these categories. The categories included antigens missed by serology, nonexpressed (null) alleles, PCR amplification failures, misassignment of antigens and nomenclature issues. Only a single individual was found to carry a null allele. DNA-based testing correctly typed nearly 99% of the donors at HLA-A, more than 98% at HLA-B, and more than 97% at both HLA-A and -B validating this methodology for registry typing.

Bone Marrow Examination↗

HLA-AB typing by polymerase-chain reaction with sequence-specific primers: more accurate, less errors, and increased resolution compared to serological typing.

Until recently, serological typing has been the primary technique used for HLA class I analysis. But because of limitations, molecular-typing techniques have replaced or supplemented the microlymphocytotoxicity test. It has been assumed that HLA class I serological typing was more accurate than serological HLA-DR typing; the latter has been shown to have 10-25% errors. But several studies have shown that HLA-AB typing was poorer than expected, and error frequencies between 5-25% were reported. This study systematically investigated the accuracy of HLA class I serological AB typing in healthy, bone-marrow registry donors, necrokidney donors, kidney-transplantation patients (on waiting lists), and haematological disorder patients. Genomic HLA class I typing, which uses polymerase-chain reaction with sequence-specific primers (PCR-SSP), gave discrepant results in 3-24% of the patients, compared to serological typings. The highest error rate (24%) was found among haematological disorder patients. Among the kidney waiting-list patients and necrokidney donors, 11% discrepancies were found. In the consecutively typed bone-marrow donors group, 3% errors were found. But among those with only one detected HLA-A specificity, 12% discrepancies were found, and among donors with only one detected HLA-B specificity, 19% errors were found. Based on these results, we recommend that patients with haematological disorders should be typed using genomic techniques. In investigations of bone-marrow registry donors and kidney patients, in which only one serological specificity is found, additional typing by genomic methods should be done.

Alleles↗

A comparison of peripheral blood coculture versus 18- or 24-month serology in the diagnosis of human immunodeficiency virus infection in the offspring of infected mothers. Women and Infants Transmission Study.

The Women and Infants Transmission Study (WITS) has established virologic definitions of human immunodeficiency virus (HIV)-infected and uninfected children that have been widely used but never formally compared with serologic definitions of infection. Data from the offspring of HIV-infected women in the WITS with frequent HIV cultures during the first year of life and with HIV serology at 18 and/or 24 months of age were analyzed. Seventy-seven infants were HIV-infected and 430 uninfected by both virologic and serologic criteria. Thirteen were virologically infected (> or = 2 positive cultures) but either seronegative or serologically indeterminate. All but 1 of these had clinical HIV disease at the time of analysis. In this pediatric cohort, children defined as infected by virologic criteria often (13/90) had negative or indeterminate serology despite symptoms of HIV disease. Results suggest that serology at 18-24 months has high specificity but poor sensitivity. It should not be considered the reference standard in identifying HIV infection in perinatally exposed children.

Blotting, Western↗

Serological study of responses to selected pathogens causing respiratory tract infection in the institutionalized elderly.

In a prospective 2-year study, serological responses to selected pathogens were analyzed in 224 episodes of fever attributable to respiratory tract infection (51.8%) or of unknown source (48.2%) in 131 residents of two long-term-care facilities. A serological response was identified in 45 episodes (20.1%): Chlamydia pneumoniae (14 episodes), Haemophilus influenzae type b (1), influenza virus type A (14), respiratory syncytial virus (RSV;2), parainfluenza virus type 3 (7), C. pneumoniae and H. influenzae (3), C. pneumoniae and influenza virus type A (2), C. pneumoniae and RSV (1), and C. pneumoniae and parainfluenza virus type 3 (1). No serological responses to Chlamydia psittaci, Chlamydia trachomatis, parainfluenza virus types 1 and 2, influenza virus type B, or Mycoplasma pneumoniae were seen. Vaccination did not affect the duration of fever in those residents with serologically confirmed influenza A. Serologically confirmed C. pneumoniae infection was detected in 9.4% of all febrile episodes. Serological responses to a second agent were detected in 33% of the patients with C. pneumoniae infections, and these dual infections were associated with an underlying malignancy (P = .02). C. pneumoniae should be recognized as a potential pathogen when choosing empirical antimicrobial therapy for respiratory tract infection in residents of long-term-care facilities.

Aged↗

The diagnostic value of streptococcal serology in early arthritis: a prospective cohort study.

OBJECTIVE: To evaluate the diagnostic value of streptococcal serology in adult early arthritis patients in discriminating between post-streptococcal reactive arthritis (PSRA) and arthritis with other causes. METHODS: The antistreptolysin-O (ASO) and anti-DNase B tests were performed at baseline in 366 consecutive, newly referred early arthritis patients. After 1 yr of follow-up the patients were classified according to international classification criteria and were evaluated for the presence of persistent arthritis. The outcome measures were the predictive value of streptococcal serology for the diagnosis of PSRA and the ability of this serology to discriminate at the first visit between the self-limiting and persistent forms of arthritis. RESULTS: With a positive serological result, the probability of having PSRA increased from 2 to 9%, whereas the probabilities of having rheumatoid arthritis or undifferentiated arthritis continued to be high (23 and 29%). The serological tests did not discriminate between the self-limiting and persistent forms of arthritis. The major Jones criteria apart from arthritis were not observed. CONCLUSION: Streptococcal serology has no diagnostic value in adult early arthritis patients in whom major Jones criteria other than arthritis are not present.

Adult↗

DNA homologies and serologic relationships among ureaplasmas from various hosts.

The available data on the serologic and genomic relationships among the established and unspeciated Ureaplasma species and serovars isolated from various hosts can be summarized as follows. Ureaplasma urealyticum (human) is composed of 14 serovars separated into two genomic clusters. Ureaplasma diversum (bovine) is antigenically complex, has three serologic clusters and requires the three representative antisera to identify all U. diversum strains. The nonhuman primate strains form four serologic groups, and each serogroup is composed of strains isolated from primates belonging to one of four distinct zoologic primate families. The ovine-caprine strains have two serologic clusters. Canine strains form four serologic clusters but serovars 1 and 2 are closely related by DNA homology. Avian strains belong to one serogroup with two genomic clusters. The DNA homology data indicate that phenotypic information alone, including antigenic serotypic data, is not always adequate for species designation among the ureaplasmas and that comparative analyses of the genome provide invaluable data for establishing new species. Although there are only two established species, the published data support the contention that the nonhuman primate, ovine-caprine, canine, feline and avian ureaplasmas are genomically and phenotypically distinct from each other based on the serologic, polyacrylamide gel electrophoresis protein and DNA cleavage patterns and DNA homology studies and that these ureaplasmas from various hosts may represent new species or subspecies within the genus.

Animals↗

Routine serologic screening for syphilis in hospitalized patients: high prevalence of unsuspected infection in the elderly.

BACKGROUND AND OBJECTIVES: To evaluate the potential yield of routine serologic testing for syphilis in hospitalized patients. STUDY DESIGN: Sera from consecutive patients admitted to an inpatient general medical service of a Baltimore hospital were tested using the RPR and FTA-ABS tests for syphilis. Two hundred ninety-one (149 males and 142 females) of 344 patients (85%) hospitalized during the study period participated, with ages ranging from 17 to 98 years. Seventy-seven percent of participants were African-American. Results of serologic tests for syphilis and human immunodeficiency virus were correlated with questionnaire responses. Associations were sought between results of serologic testing and demographic characteristics, medical problems, and a history of syphilis. RESULTS: No patient had an admitting diagnosis of syphilis. Thirty-seven patients (13%) had both reactive RPR and FTA-ABS tests. Although serologic evidence of syphilis was significantly associated with age older than 50 years and with a history of syphilis, 59% of patients with both reactive RPR and FTA-ABS tests did not give a history of syphilis. Ten percent of patients were HIV seropositive; however HIV seropositivity was not significantly associated with serologic evidence of syphilis. CONCLUSION: Unsuspected latent syphilis was common in this population. Routine serologic screening for syphilis should be considered in some hospitals.

Adolescent↗

HLA-DR and DQ typing by polymerase chain reaction using sequence-specific primer mixes reduces the incidence of phenotypic homozygosity (blanks) over serology.

Because of the inherent difficulties in allele assignment with HLA-DR serological typing, in 1993 our organ procurement organization-based HLA laboratory replaced serology with the molecular method of polymerase chain reaction using sequence-specific primer mixes (PCR-SSP) to type for DR and DQ at a resolution level equivalent to that of serologically defined antigens. In this study, we compared the incidence of DR blanks, where allocative homozygosity occurred, and graft outcome during our serology epoch (1987-1993) with that of our molecular epoch (1993-1996). The incidence of DR blanks by PCR-SSP (17.0%; 138/1101) was significantly lower (P<0.005) than in the serology epoch (21.5%; 569/2647). Although DQ is not a component of the allocation algorithm, the incidence of blanks in the molecular era (21.9%; 196/895) was 46% lower (P<0.001) than in the serology epoch (40.8%; 931/2277). Graft survival in 163 cadaveric renal transplant recipients for whom molecular DR allocation occurred (patient and donor were molecularly typed) showed that PCR-SSP typing had no significant effect on 2.5-year graft survival for patients mismatched for 0 (97%), 1 (90%), or 2 (94%) HLA-DR antigens (P=0.4; log-rank). In conclusion, molecular typing lowered the rate of DR and DQ blanks, but molecular matching for HLA DR and DQ did not influence graft outcome at 2.5 years.

Blood Grouping and Crossmatching↗

Serological viral testing of cadaveric cornea donors.

BACKGROUND: Cornea graft recipients are exposed to viral transmission from the donor. Cadaveric donor serum is often of poor quality and frequently yields falsely positive results in serological assays that may result in the graft being needlessly discarded. OBJECTIVE: We examined the influence of the time of blood collection after death, and the macroscopic aspect of serum, on serological test results in cadaveric cornea donors. METHODS: Five hundred sixty-five consecutive cadaveric cornea donors were systematically tested for serological markers of human immunodeficiency virus type 1 and 2, human T-cell leukemia virus type 1, hepatitis B and hepatitis C viruses (HCV). We studied the influence of the macroscopic aspect of the donor's serum and the time of blood collection after death on the results of serological testing and on the subsequent decision to use or discard the graft. RESULTS: Twenty-one and a half percent of corneas were rejected on the basis of virological test results. We found significant relationships between the macroscopic aspect of serum at the time of testing and: (i) a positive, equivocal or discrepant result of immunoassays, for all markers except anti-HCV antibodies, (ii) non acceptance of cornea grafts, and (iii) the time of blood sampling after death. CONCLUSIONS: The macroscopic aspect of postmortem blood samples is the best predictor of the specificity of serological testing in cornea donors. Serological results should be interpreted with care when serum is macroscopically abnormal, and cadaveric donors should not be sampled more than 12 hr after death.

Cadaver↗

Comparison of two serological methods and a polymerase chain reaction-enzyme immunoassay for the diagnosis of acute respiratory infections with Chlamydia pneumoniae in adults.

Chlamydia pneumoniae is a common respiratory tract pathogen. Serological methods currently used for the diagnosis of C. pneumoniae infection lack specificity, give ambiguous results from a single serum sample and often provide only a retrospective diagnosis. A prospective study was undertaken to assess whether PCR could be a useful addition to the serological techniques routinely practised for diagnosis. This study investigated 68 adult patients with a diagnosis of acute respiratory infection. Acute and convalescent serological determination of antibodies to C. pneumoniae were performed by means of an rELISA test and a micro-immunofluorescence (MIF) test. Nasopharyngeal aspirates or bronchoalveolar lavage specimens and bronchial aspirates obtained from the 68 patients were evaluated by PCR-enzyme immunoassay (PCR-EIA) for the presence of C. pneumoniae and by immunofluorescence assay and cell culture for virus identification. Mycoplasma pneumoniae serology was also performed. Eight patients (11.8%) were positive by either rELISA or PCR-EIA, or both, with an infection rate of 5 (18.5%) of 27 in patients with community-acquired pneumonia, 2 (9%) of 22 in asthmatic patients and 1 (5%) of 19 in patients with an exacerbation of chronic obstructive pulmonary disease. Serological evidence of acute infection was found in four of these patients with the rELISA test and in three others with the MIF test. PCR-EIA detected C. pneumoniae DNA in four specimens, but there were concordant results with both rELISA and PCR-EIA in only one patient A positive PCR-EIA was also obtained in a patient who did not show an antibody response in acute serum. The discrepancy between serological and PCR-EIA results reflects the difficulties in routine laboratory diagnosis of C. pneumoniae infection and the necessity for further studies with optimised techniques.

Acute Disease↗

The application of serological techniques to the taxonomy of Arthrobacter and related organisms.

Antisera were raised against rods of 17 named Arthrobacter and Aureobacterium strains. Antigenic relationships between these strains, other soil bacteria and new Arthrobacter isolates from several soils were studied, using agglutination, immunodiffusion, immunofluorescence and ELISA techniques. Many of the named Arthrobacter species had common antigens, and there were also common antigens amongst named Arthrobacter strains and many fresh Arthrobacter isolates. Agglutination, ELISA and immunofluorescence tests revealed greater antigenic differences between the named strains than did immunodiffusion tests. Serological similarities between the 17 named strains and the fresh Arthrobacter isolates were calculated using SJ coefficients. The occurrence of named strains in serogroups based on immunodiffusion data supported the taxonomic scheme for arthrobacters in Bergey's Manual of Systematic Bacteriology. The distribution of soil isolates in serogroups resembled that in groups based on numerical analysis of diverse characters. This makes it possible to use serological tests to locate particular species in soil samples. Arthrobacter atrocyaneus was serologically distinct from other members of the Ar. globiformis/Ar. citreus group and did not cluster with them phenetically. Serological data suggest that Ar. aurescens, Ar. ureafaciens and Ar. histidinolovorans constitute a single species. Although A. simplex and A. tumescens have been placed in the genus Pimelobacter, they are serologically distinct from one another and more closely resemble Ar. globiformis. Aureobacterium strains were serologically distinct from arthrobacters. Micrococcus roseus showed many cross-reactions with several antisera, supporting the placement of micrococci in the same family as arthrobacters.

Animals↗

The serology of Bordetella bronchiseptica isolated from pigs compared with strains from other animal species.

The serology of freshly isolated strains of Bordetella bronchiseptica from pigs and some other animal species was studied by the agglutination and agglutinin absorption tests. Differentiation was made between heat stable O-antigens and heat labile K-antigens. All the strains examined contained a common O-antigen (1). In addition, three heat stable antigens were detected. The heat labile antigens of freshly isolated strains were found to be very different. The major antigens 2 and 3 were not found in combination. A common weak K-antigen (1) was found in all strains. The pig strains were found to be serologically identical (0, 1, 2; K 1, 2). The same serotype was isolated from a rabbit. Another rabbit strain was found to be similar to a strain isolated from an SPF-rat (O, 1, 3; K 1, 3, 4). The antigenic pattern found in two cat strains was O 1, 3; K 1, 3. Typing of freshly isolated strains from pigs with monospecific factor sera confirmed that all the strains so far examined were serologically identical. This uniform serology hampers epidemiological studies of porcine bordetellosis by antigenic analysis. The occurrence of serologically different serotypes of B. bronchiseptica is of significance in the diagnostic bacteriology since the final diagnosis must be confirmed by a serological test.

Agglutination Tests↗

Effect of feline immunodeficiency virus infection on Toxoplasma gondii-specific humoral and cell-mediated immune responses of cats with serologic evidence of toxoplasmosis.

Serum samples from 89 cats with serologic evidence of toxoplasmosis were identified by using an enzyme-linked immunosorbent assay (ELISA) that detected Toxoplasma gondii-specific immunoglobulin M (IgM) or T. gondii-specific immunoglobulin G (IgG). Concurrent feline immunodeficiency virus (FIV) infection was detected in 36 cats using an ELISA for detection of FIV-specific IgG. The majority of the cats in both the FIV-seropositive and FIV-seronegative groups were male and > 5 years of age. FIV-seropositive cats were more likely to have T. gondii IgM titers without IgG (P < 0.05) or any T. gondii IgM titer (P < 0.05) than were FIV-seronegative cats. FIV-seronegative cats (1328) had a higher T. gondii IgG geometric mean titer than did FIV-seropositive cats (724) and were more likely to have T. gondii IgG titers > 1:2048 than were FIV-seropositive cats (P < 0.05). Cats with serologic evidence of both T. gondii and FIV infections had persistent T. gondii IgM titers for > 12 weeks. Lymphoblast transformation in response to concanavalin A, T. gondii-specific intracellular antigens, and T. gondii-specific secretory antigens was compared in T. gondii seropositive and FIV-seronegative cats, cats with serologic evidence of T. gondii infection alone, and cats with serologic evidence of concurrent FIV and T. gondii infections. Lymphocytes from all but one cat in the FIV-seropositive group responded to concanavalin A. Whereas lymphocytes from FIV-seronegative cats with serologic evidence of toxoplasmosis responded to T. gondii-specific antigens, four of five of the FIV-seropositive cats with concurrent serologic evidence of toxoplasmosis did not.

Animals↗

Reliability of Helicobacter pylori and CagA serological assays.

Background serological assays for Helicobacter pylori are commonly used without knowledge of reliability. This information is needed to define the ability of serological tests to determine either new cases of infection or loss of infection in longitudinal studies. We evaluated the reproducibility and the interrelationships of serological test results for H. pylori and cytotoxin-associated gene product A (CagA) enzyme-linked immunoassays within a subset of participants in a population-based study. Stored samples from 1,229 participants in the third U.S. National Health and Nutrition Examination Survey were replicate serologically tested for H. pylori and CagA. Overall disagreement was 3.4% between duplicate tests for H. pylori (or 2.3% if equivocal results were disregarded). Six percent of samples positive on the first test had an immune serum ratio at least 30% lower on repeat testing. The odds ratio for H. pylori seropositivity on retesting was 2.8 (95% confidence interval [CI] = 1.8 to 4.5) when CagA serology was positive versus when it was negative. CagA antibody was found among 47.8% of H. pylori-equivocal and 7.0% of H. pylori-negative samples. CagA-positive yet H. pylori-negative samples were more likely to occur among Mexican Americans (odds ratio, 5.2; 95% CI = 2.4 to 11.4) and non-Hispanic blacks (odds ratio, 5.5; 95% CI = 2.3 to 13.0) than among non-Hispanic whites. Relying on repeated H. pylori serological tests over time to determine infection rates may result in misinterpretation due to limits in test reproducibility. CagA testing may have a role in verifying infection.

Adult↗

SEROLOGY AND TRANSDUCTION IN STAPHYLOCOCCAL PHAGE.

Dowell, C. E. (The University of Texas, Dallas) and E. D. Rosenblum. Serology and transduction in staphylococcal phage. J. Bacteriol. 84:1071-1075. 1962.-A triply lysogenic strain of Staphylococcus aureus was shown to carry a serological group B phage capable of transduction. Three typing phages (53, 80, 42D), either belonging to serological group B or having a close association with it, were also shown to have transducing ability. A rapid screening method was used to isolate two new transducing phages, both of which belonged to serological group B. Propagating strain 42B/47C was found to carry a transducing phage that was neutralized by both group B and group F antisera. Nine other phages belonging to serological groups other than group B did not have generalized transducing ability, nor did three group B typing phages that were atypical in their calcium requirement. It was postulated that transducing ability is associated with staphylococcal phages of serological group B and with related phages of group F.

Journal Article↗

Cell wall composition and serological reactivity of Histoplasma capsulatum serotypes and related species.

The composition of the cell walls of strains of Histoplasma capsulatum that were reactive in the complement-fixation test and that reacted with fluorescein-tagged specific anti-H. capsulatum serum were compared with the cell walls of H. capsulatum and Histoplasma duboisii that did not react in these serological tests. The cell walls of the serologically reactive strains of H. capsulatum were much more susceptible to the action of proteinases and to chitinase than were the serologically nonreactive strains, but were less susceptible to hydrolysis by 3% NaOH or 2 n H(2)SO(4). The walls of the serologically reactive strains had higher chitin, amino acid, and hexosamine contents, but they were lower in a "lactic acid" component that was released by alkaline hydrolysis of the nonreactive strains. The cell walls of strains of H. duboisii were similar to the walls of the serologically nonreactive strains of H. capsulatum. Chemical analyses of the cell walls of the various strains clearly support the serological typing of H. capsulatum and H. duboisii strains reported by Kaufman and Blumer.

Animals↗

Serological and virological characterization of clinically diagnosed cases of measles in suburban Khartoum.

Measles continues to be a major childhood disease in terms of global morbidity and mortality. In the main areas of its endemicity the only available means of diagnosis are based on clinical criteria: the presence of a maculopapular rash and fever accompanied by cough, coryza, and/or conjunctivitis. We have studied 38 clinically diagnosed cases of measles in Khartoum, Sudan, by means of serology, reverse transcriptase PCR (RT-PCR) on throat swabs and virus isolation from lymphocytes. On the basis of serology, 28 patients were diagnosed as having an acute measles virus (MV) infection, while in 10 cases the clinical symptoms proved to have other causes. It was shown that in cases with low serum immunoglobulin M (IgM) levels, an additional measurement of IgG or virus-neutralizing antibodies was necessary to discriminate between patients with an acute MV infection sampled during an early stage of the disease and patients who had experienced an MV infection in the more distant past. The serological laboratory diagnosis was validated by an MV-specific RT-PCR: for all confirmed measles cases tested a fragment of the correct size which hybridized with a third MV-specific primer could be amplified, while all serologically negative cases were also RT-PCR negative. MV could be isolated from 17 out of 23 of the serologically confirmed cases, demonstrating that virus isolation is less reliable as a diagnostic tool than serology or RT-PCR. This study stresses the urgent need for a rapid diagnostic field test for measles.

Adolescent↗