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V Pope

Publications and source records attributed to V Pope.

29 records · Page 2Linked to original sources

Effect of heat or formalin treatment of leptospires on antibody response detected by immunoblotting.

Leptospira interrogans serovar icterohaemorrhagiae RGA (RGA), live or heated at 56 degrees C for 15 min or treated with Formalin, was injected into rabbits to prepare hyperimmune serum. The pathogens L. interrogans serovars icterohaemorrhagiae RGA, icterohaemorrhagiae 1, canicola Moulton, grippotyphosa Andaman, hardjo Hardjoprajitno, and pomona Pomona and the nonpathogen Leptospina biflexa serovar patoc Patoc 1 were processed for sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and after electrophoresis they were then transferred to nitrocellulose paper. Antiserum against RGA (live, heat killed, or Formalin killed) was used on one of each of the three blots. Formalin appeared to completely eliminate antibody response to antigens with the molecular weight of 14,000 to 20,000 (14K to 20K) but did expose an antigen at approximately 23K in the pathogens only. This same band had only slight reactivity when antiserum against heat-killed RGA was used. Heating also eliminated cross-reactivity in the 19K to 30K range and partially degraded bands in the 14K to 20K region so that one broad band rather than several discrete bands appeared. The three antiserum specimens cross-reacted with all of the serovars tested, but fewer antigens of grippotyphosa and hardjo reacted with the antisera. Against patoc, reactivity was limited primarily to the flagellar region. The most cross-reactivity was with the antiserum prepared by using live leptospires.

Animals↗

Effect of heat or chemical treatment on leptospiral antigens.

The use of bacterins is one of the primary methods in the control of leptospirosis in domestic animals, especially cattle, swine, and dogs. Bacterins have been made using chemically treated or heat-inactivated organisms. Many of the initial studies indicated that the bacterins gave complete protection, whereas later reports found that some inactivation methods resulted in bacterins that protect against death but not against kidney infection and renal shedding. This raised the possibility that some of the antigens had been altered. To investigate this possibility, we heat or chemically treated Leptospira interrogans serovar icterohaemorrhagiae CF-1, L. interrogans serovar canicola Moulton, L. interrogans serovar grippotyphosa SC4397, L. interrogans serovar hardjo Hardjoprajitno, and L. interrogans serovar pomona MLS. Temperatures used to determine alterations in antigens were 50, 56, 80, 100, and 121 degrees C. The chemicals used were formaldehyde, phenol, and thimerosal. Analysis was done using Laemmli gels and Western blots. We found that heating at 50 or 56 degrees C had the smallest effect on antigens, whereas heating at 80 or 100 degrees C caused the appearance of high-molecular-weight bands on Western blots. Heating at 121 degrees C caused almost all the bands to disappear, in both the stained gel and the Western blot. Both phenol and Formalin treatment altered proteins, whereas thimerosal treatment appeared to have little effect compared with heating at 50 degrees C.

Antigens, Bacterial↗

Unheated serum reagin test as a quantitative test for syphilis.

The unheated serum reagin (USR) test, a standard qualitative test for syphilis, was evaluated for use as a quantitative test for the serodiagnosis of syphilis and as a means of determining the effectiveness of treatment. Tests were performed on 664 syphilitic sera from treated and untreated patients in the primary, secondary, and latent stages of the disease. Also included were 95 false-positive sera and 36 sera from presumably normal donors. The Venereal Disease Research Laboratory slide and rapid plasma reagin 18-mm circle card tests were used as a basis for comparison with the USR test. Results of the USR quantitative test paralleled the Venereal Disease Research Laboratory and rapid plasma reagin card quantitative tests with approximately 84% agreement. The advantages of the USR test make the findings of this study significant in that: (i) less technical manipulation is required to perform the USR test than the Venereal Disease Research Laboratory slide test; (ii) no labor cost is incurred for daily Venereal Disease Research Laboratory antigen preparation; and (iii) the USR antigen is less expensive than the rapid plasma reagin card tests antigen.

Antibodies↗

Evaluation of the microenzyme-linked immunosorbent assay with Treponema pallidum antigen.

Whole-cell sonicates of Treponema pallidum, Nichols strain, were evaluated in an enzyme-linked immunosorbent assay (Elisa) for syphilis, and results were read in a Dynatek Microelisa Reader. The antigen was evaluated with sera from patients with syphilis, persons presumed normal, and biological false-positives. Two hundred and ninety-seven sera were tested by the ELISA with T. pallidum antigens, the Venereal Disease Research Laboratory (VDRL) slide test, the fluorescent treponemal antibody absorption (FTA-Abs) test, and the microhemagglutination assay for T. pallidum antibodies (MHA-TP). The results of all of the tests were compared. The ELISA, with 89.3% sensitivity, was less sensitive than the VDRL (93.3%) and FTA-Abs (100.0%) tests but more sensitive than the MHA-TP (76.0%). THe ELISA was considerably more sensitive in primary syphilis than the MHA-TP. Specificity was as follows: ELISA, 98.5%; FTA-Abs test, 97.8%; MHA-TP, 98.2%; and VDRL test, 92.7%. The ELISA has good potential as a confirmatory test in the serodiagnosis of syphilis.

Antibodies, Bacterial↗

Blinded proficiency testing of FTA-ABS test.

In 1976 and 1977, 28 state laboratories participated in a blinded proficiency testing study of syphilis serologic tests. The test considered of a battery of 50 duplicate reagin-positive sera; 12 specimens were obtained from syphilitic patients. The median reproducibility scores for the VDRL and FTA-ABS tests were the same, 92%, and the scores for the two tests were correlated. Given the relatively low prevalence of untreated syphilis in the United States today, serologic results that are highly predictive of syphilis are obtainable only from the most proficient laboratories, if unselected sera are tested. Reactivity in the VDRL and FTA-ABS tests does not prove the diagnosis of syphilis. Physicians must use clinical judgment to preselect patients for serologic testing.

Laboratories↗

Center for Disease Control Diagnostic Immunology Proficiency Testing Program results for 1978.

Data from about 1,000 laboratories participating in the Diagnostic Immunology portion of the 1978 Center for Disease Control Proficiency Testing Program provided information dealing with laboratory performance and trends in testing protocols. Ninety specimens were distributed in scheduled quarterly and semiannual shipments, and five additional specimens were provided in a special survey. The specimens offered both qualitative and quantitative challenges for a wide variety of analytes which included syphilis serology, rheumatoid factor, bacterial agglutinins, hepatitis B surface antigen, immunoglobulins and other serum proteins, infectious mononucleosis, rubella, toxoplasma, antinuclear antibodies, and streptococcal exoenzymes. This paper summarizes the results of the 1978 program.

Agglutinins↗

Center for Disease Control Diagnostic Immunology Proficiency Testing Program results for 1977.

A summary of the yearly data accumulated and analyzed in the diagnostic immunology portion of the Center for Disease Control 1977 Proficiency Testing Program provides information on the trends in testing protocols from over 900 participating laboratories. One hundred and fourteen specimens prepared by the Center for the Disease Control were distributed quarterly or in special surveys. The specimens chosen provided feedback from a broad range of tests commonly performed in diagnostic immunology laboratories and included those for rubella antibodies, hepatitis B surface antigen, bacterial antibodies, rheumatoid factor, immunoglobulins and other serum-specific proteins, syphilis serology, and carcinoembryonic antigen. This summary provides an overview of the trends observed since last year's report.

Centers for Disease Control and Prevention, U.S.↗

Center for disease control diagnostic immunology proficiency testing program results for 1976.

Over 900 laboratories participated in the Diagnostic Immunology portion of the 1976 Proficiency Testing Program, which was provided by the Center of Disease Control under the authority of the Clinical Laboratories Improvement Act of 1967. One hundred specimens prepared by the Center for Disease Control for analysis were distributed on a quarterly schedule or in special surveys. Feedback from participating laboratories included over 37,500 qualitative and 33,000 quantitative responses, which were analyzed to determine individual laboratory proficiency levels. In addition, information supplied by participants in each survey helped to delineate trends in testing protocols. The specimens chosen for analysis called for a broad range of tests commonly performed in diagnostic immunology laboratories, including those for rubella antibodies, hepatitis B surface antigen, bacterial antibodies, rheumatoid factor, immunoglobulins and other serum-specific proteins, and carcinoembryonic antigen. A summary of the data analysis is provided so that the laboratories can improve their overall performance levels.

Agglutination Tests↗

Four serologic tests for syphilis: results with comparison of selected groups of sera.

The specificity of the fluorescent treponemal antibody-absorbed (FTA-Abs) test was assessed for 17 sera from syphilitic patients that were nonreactive in the Treponema pallidum immobilization (TPI) test but reactive in the FTA-Abs test. Thirty-three other sera from syphilitic patients and 19 sera from nonsyphilitic individuals were also examined by fluorescent treponemal and microhemagglutination Treponema pallidum (MHA-TP) tests and by the enzyme-linked immunosorbent assay (ELISA). Specific absorptions of sera with calf thymus DNA or Treponema pallidum biotype Reiter (Reiter treponemes) were performed. In quantitative immunofluorescence assays (IFA) with antihuman IgG and IgM conjugates, results were similar to those for reactive sera from a control group. Results of both the MHA-TP and ELISA tests supported the specificity of the FTA-Abs test; reactivity in the latter was not removed by specific absorption either with calf thymus DNA or with Reiter treponemes. This evaluation suggests a format for serodiagnosis in cases in which test results are discrepant.

Enzyme-Linked Immunosorbent Assay↗

Testing umbilical cords for funisitis due to Treponema pallidum infection, Bolivia.

To establish the frequency of necrotizing funisitis in congenital syphilis, we conducted a prospective descriptive study of maternal syphilis in Bolivia by testing 1,559 women at delivery with rapid plasma reagin (RPR). We examined umbilical cords of 66 infants whose mothers had positive RPR and fluorescent treponemal antibody absorption tests. Histologic abnormalities were detected in 28 (42%) umbilical cords (seven [11%] had necrotizing funisitis with spirochetes; three [4%] had marked funisitis without necrosis; and 18 [27%] had mild funisitis), and 38 [58%] were normal. Of 22 umbilical cords of infants from mothers without syphilis (controls), only two (9%) showed mild funisitis; the others were normal. Testing umbilical cords by using immunohistochemistry is a research tool that can establish the frequency of funisitis due to Treponema pallidum infection.

Antibodies, Bacterial↗