scholarly journals Sensitive Enzyme-Linked Immunosorbent Assay for Detection of Antibodies Against Typhus Rickettsiae, Rickettsia prowazekii and Rickettsia typhi

1977 ◽  
Vol 6 (2) ◽  
pp. 101-110
Author(s):  
Sidney Halle ◽  
Gregory A. Dasch ◽  
Emilio Weiss
Keyword(s):  
Immunosorbent Assay ◽  
Complement Fixation ◽  
Differential Susceptibility ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rickettsia Prowazekii ◽  
Rickettsia Typhi ◽  
Titration Curves ◽  
Ether Extraction ◽  
Human Sera

An enzyme-linked immunosorbent assay (ELISA) has been developed for the titration of rickettsial antibodies in human and animal sera. Two preparations of soluble typhus-group antigens were obtained from Rickettsia typhi and Rickettsia prowazekii by ether extraction: a standard antigen from infected yolk sacs (YS antigen) and one free of yolk sac contaminants from Renografin-purified rickettsiae (PR antigen). Rabbit, mouse, and guinea pig sera were obtained by immunization with viable purified R. typhi or R. prowazekii . Human sera were obtained from individuals who had recovered from laboratory infections with either typhus rickettsia months or years previously. Goat-derived anti-immunoglobulins were conjugated to alkaline phosphatase with glutaraldehyde. Although the PR and YS antigens gave equivalent antibody titers in the complement fixation test, the PR antigen was clearly superior in the ELISA. With this antigen, the titration curves of all antisera were linear over a wider range of serum concentrations and the titers were higher than with the YS antigen. With YS and PR antigens, ELISA titers were higher than those obtained by complement fixation by one and two orders of magnitude, respectively. In human sera, immunoglobulin G and immunoglobulin M antibodies were demonstrated by their respective anti-immunoglobulins and by differential susceptibility to ethanethiol. ELISA titers showed some type specificity, whereas none was observed in complement fixation tests. The ELISA is highly sensitive, reproducible, and easily adaptable to the various requirements of clinical and research laboratories.

scholarly journals Lateral-Flow Assay for Rapid Serodiagnosis of Human Leptospirosis

2001 ◽  
Vol 8 (1) ◽  
pp. 166-169 ◽  
Author(s):  
Henk L. Smits ◽  
C. K. Eapen ◽  
Sheela Sugathan ◽  
Mariamma Kuriakose ◽  
M. Hussein Gasem ◽  
...  
Keyword(s):  
Positive Result ◽  
Immunosorbent Assay ◽  
Rapid Detection ◽  
Test Line ◽  
Lateral Flow ◽  
Immunoglobulin M ◽  
Lateral Flow Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Immunoglobulin ◽  
Human Sera

ABSTRACT An assay device for the rapid detection ofLeptospira-specific immunoglobulin M (IgM) antibodies in human sera is presented. The sensitivity (85.8%) and specificity (93.6%) of the assay compared well (91.9% agreement) with those of an IgM enzyme-linked immunosorbent assay routinely used in the serodiagnosis of leptospirosis. The sensitivity of the assay varied with the stage of the disease. The assay uses stabilized components and is simply performed by the addition of serum and sample fluid to the sample well of the assay device. The assay is read after 10 min, and a positive result is obtained when staining of the test line is observed.

scholarly journals Comparison of a Commercial Enzyme-Linked Immunosorbent Assay with Immunofluorescence and Complement Fixation Tests for Detection of Coxiella burnetii (Q Fever) Immunoglobulin M

2000 ◽  
Vol 38 (4) ◽  
pp. 1645-1647 ◽  
Author(s):  
Peter R. Field ◽  
Jody L. Mitchell ◽  
Avelina Santiago ◽  
David J. Dickeson ◽  
Sau-Wan Chan ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Coxiella Burnetii ◽  
Complement Fixation ◽  
Q Fever ◽  
Fluorescent Antibody ◽  
Reference Method ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Igm Elisa

A commercially available enzyme-linked immunosorbent assay (ELISA) for the diagnosis of Q fever (PanBio Coxiella burnetiiimmunoglobulin M [IgM] ELISA, QFM-200) was compared to the indirect fluorescent antibody test (IFAT) for C. burnetii IgM and the complement fixation test (CFT). The ELISA demonstrated 92% agreement with the reference method (IFAT), and gave a sensitivity of 99% (69 of 70 samples) and a specificity of 88% (106 of 121). Specificity can be increased with confirmation by IFAT. CFT was found to have a specificity of 90% (107 of 119), although it was lacking in sensitivity (73%; 51 of 70). No cross-reactivity was observed in the ELISA with serum samples from patients with mycoplasma (n = 6), chlamydia (n = 5), or legionella (n = 4) infections, although 2 of 5 patients with leptospirosis and 1 of 4 samples containing rheumatoid factor (RF) demonstrated positive results in the ELISA. Results indicate that the performance of the PanBio C. burnetii (Q fever) IgM ELISA (F = 187) is superior to that of CFT (F = 163), and consequently the ELISA should be a useful aid in the diagnosis of acute Q fever.

scholarly journals Enzyme-linked immunosorbent assay for chlamydial antibodies

1977 ◽  
Vol 6 (5) ◽  
pp. 507-510
Author(s):  
V J Lewis ◽  
W L Thacker ◽  
S H Mitchell
Keyword(s):  
Immunosorbent Assay ◽  
Chlamydial Infection ◽  
Complement Fixation Test ◽  
Complement Fixation ◽  
Chlamydia Psittaci ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Sera ◽  
History Of ◽  
Species Specific ◽  
Formalin Fixed

An enzyme-linked immunosorbent assay (ELISA) detected chlamydial antibodies in human sera. The assay antigen produced in cell cultures infected with Chlamydia psittaci was Formalin-fixed to microplates. Single convalescent-phase sera positive for chlamydial antibodies by a complement-fixation test were positive at even higher dilutions by ELISA. Paired sera with diagnostic rises in complement-fixing antibody showed seroconversion by ELISA also. Control sera from persons with no history of chlamydial infection were negative by both tests. Sera from patients with psittacosis or lymphogranuloma venereum were ELISA positive, indicating that the assay with the antigen used in this study is genus specific rather than species specific.

scholarly journals Combined Cell Culture Enzyme-Linked Immunosorbent Assay for Quantification of Poliovirus Neutralization- Relevant Antibodies

2000 ◽  
Vol 7 (6) ◽  
pp. 915-919 ◽  
Author(s):  
A. F. Wahby
Keyword(s):  
Cell Culture ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Humoral Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
Titration Curves ◽  
Microneutralization Assay ◽  
Human Sera ◽  
Reference Serum ◽  
Presence And Absence

ABSTRACT A combined cell culture enzyme-linked immunosorbent assay (CCC-ELISA) was developed for measuring the neutralizing antipoliovirus antibodies in human sera. The binding of different concentrations of each of the three poliovirus types to BGM cells in the presence and absence of a constant dilution from each test and reference serum was measured in the CCC-ELISA. The titers of the viruses neutralized by each serum were measured with the titration curves and used for interpretation of neutralizing titers to the three poliovirus types. Analysis of human sera revealed that the sensitivity and specificity of the CCC-ELISA and the microneutralization assay were comparable. The CCC-ELISA is nonsubjective, rapid, and highly reproducible. Furthermore, the CCC-ELISA could potentially be used as a seroepidemiologic tool for assessment of the humoral response to the cell culture infectious viruses.

scholarly journals Limited Diagnostic Capacities of Two Commercial Assays for the Detection of Leptospira Immunoglobulin M Antibodies in Laos

2006 ◽  
Vol 13 (10) ◽  
pp. 1166-1169 ◽  
Author(s):  
Stuart D. Blacksell ◽  
Lee Smythe ◽  
Rattanaphone Phetsouvanh ◽  
Michael Dohnt ◽  
Rudy Hartskeerl ◽  
...  
Keyword(s):  
Diagnostic Accuracy ◽  
Immunosorbent Assay ◽  
Gold Standard ◽  
Agglutination Test ◽  
Immunoglobulin M ◽  
Microscopic Agglutination Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diagnostic Utility

ABSTRACT The diagnostic utility of immunochromatographic (Leptotek) and enzyme-linked immunosorbent assay (ELISA; Panbio) tests for the detection of Leptospira immunoglobulin M antibodies was assessed in febrile adults admitted in Vientiane, Laos. Both tests demonstrated poor diagnostic accuracy using admission serum (Leptotek sensitivity of 47.3% and specificity of 75.5%: ELISA sensitivity of 60.9% and specificity of 65.6%) compared to the Leptospira “gold standard” microscopic agglutination test.

Comparison of an enzyme-linked immunosorbent assay for quantitation of rotavirus antibodies with complement fixation in an epidemiological survey

1978 ◽  
Vol 8 (3) ◽  
pp. 268-276
Author(s):  
L H Ghose ◽  
R D Schnagl ◽  
I H Holmes
Keyword(s):  
Immunosorbent Assay ◽  
Complement Fixation ◽  
Age Groups ◽  
Enzyme Reaction ◽  
Epidemiological Survey ◽  
Enzyme Linked Immunosorbent Assay ◽  
Marker Enzyme ◽  
Aminosalicylic Acid ◽  
Antibody Titers ◽  
Antibody Levels

The development of a micro-scale enzyme-linked immunosorbent assay (ELISA) with horseradish peroxidase as the marker enzyme for the detection and measurement of human rotavirus antibodies is described. A semipurified preparation of the serologically related simian agent, SA-11 virus, was used as the antigen. Test sera were reacted with antigen-sensitized wells in disposable poly-vinyl microplates. Any attached antibody was detected by the addition of peroxidase-labeled anti-species immunoglobulin (conjugate) followed by assay of the enzyme reaction with its substrate, hydrogen peroxide plus 5-aminosalicylic acid. This micro-ELISA was compared with complement fixation in a seroepidemiological study of the age prevalence of rotavirus antibody in Aboriginal and European populations living in the same outback area in Australia. The ELISA (results read with the naked eye) proved to be approximately 16 times more sensitive than complement fixation. Of Aborigines, 71% had rotavirus complement-fixing antibody, as compared to 45% of Europeans. By ELISA 100% of both populations had rotavirus antibodies. Mean antibody titers in the different age groups were higher in Aborigines than in Europeans. Antibody levels rose steeply throughout the first 20 years of life, remained high during the next 20 years, then increased again at least up to the age of 60 years. The micro-ELISA was practical, simple to perform, and more suitable than complement fixation for large seroepidemiological rotavirus studies. It also has potential for serodiagnosis of the disease, both in the laboratory and in the field.

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