scholarly journals Optimization of a Human Papillomavirus-Specific Enzyme-Linked Immunosorbent Assay

2002 ◽  
Vol 9 (3) ◽  
pp. 577-582 ◽  
Author(s):  
Kevin L. Karem ◽  
Alysia C. Poon ◽  
Cynthia Bierl ◽  
Rosane Nisenbaum ◽  
Elizabeth Unger
Keyword(s):  
Human Papillomavirus ◽  
Immunosorbent Assay ◽  
Critical Evaluation ◽  
Negative Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Optimal Cutoff ◽  
Optimal Method ◽  
Specific Immunoglobulin ◽  
Human Sera ◽  
Evaluation Techniques

ABSTRACT A strategy was developed for the control, standardization, and critical evaluation of an enzyme-linked immunosorbent assay (ELISA) for the detection of human papillomavirus-specific immunoglobulin G in human sera. Control human sera, polyclonal animal sera, and monoclonal antibodies were used to establish optimal assay parameters, including antigen coating, serum dilutions, and criteria for daily reproducibility, monitoring, and rejection of assays. Three evaluation techniques were used in parallel to define an optimal cutoff absorbance value that yields greater than 93% sensitivity and 98.5% specificity in the assay's ability to discriminate positive and negative control sera. This strategy provides an optimal method by which to determine cutoff absorbance values for ELISA.

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.

Enzyme-linked immunosorbent assay, immunoblot analysis and RAST fluoroimmunoassay analysis of serum responses against crude larval antigens of Anisakis simplex in a Spanish random population

1996 ◽  
Vol 70 (4) ◽  
pp. 281-289 ◽  
Author(s):  
L. García-Palacios ◽  
M.L. González ◽  
M.I. Esteban ◽  
E. Mirabent ◽  
M.J. Perteguer ◽  
...  
Keyword(s):  
Optical Density ◽  
Immunoblot Analysis ◽  
Specific Ige ◽  
Negative Control ◽  
Anisakis Simplex ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Sera ◽  
Specific Ige Antibodies ◽  
Reference Serum ◽  
Diagnostic Criterion

AbstractThe results obtained in a study of seroprevalence by means of ELISA and immunoblot with crude larval extracts of Anisakis simplex using 1008 human sera from Spanish people showing no clinical suspicion of anisakidosis are given. For the evaluation of the results obtained by ELISA the Diagnostic Index (DI) was used, as the ratio between the optical density resulting from the test serum and the optical density of the negative control. Forty-seven sera showed DIs between 1.5 and 2, and 14 sera were greater than 2. After comparison of the immunoblot analysis with the immunorecognition pattern of a human anisakidosis reference serum, a diagnostic criterion could be established for those sera that, at a 1/100 dilution, showed a DI by ELISA greater than 1.5. Seven of 14 selected sera with DIs in ELISA higher than 1.3 showed anti-Anisakis specific IgE antibodies by RAST fluoroimmunoassay.

scholarly journals Detection of Human Papillomavirus Type 31-Neutralizing Antibodies from Naturally Infected Patients by an Assay Based on Intracellular Assembly of Luciferase-Expressing Pseudovirions

2007 ◽  
Vol 15 (1) ◽  
pp. 172-175 ◽  
Author(s):  
Maxime J. J. Fleury ◽  
Antoine Touzé ◽  
Silvia de Sanjosé ◽  
F. Xavier Bosch ◽  
Joellen Klaustermeiyer ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Human Papillomavirus ◽  
Immunosorbent Assay ◽  
Neutralizing Antibodies ◽  
Neutralization Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Highly Sensitive

ABSTRACT The aim of this study was to develop a highly sensitive human papillomavirus type 31 (HPV31) neutralization assay based on the production of pseudovirions carrying luciferase. Neutralizing antibodies against HPV31 were investigated in a set of HPV31 monoclonal antibodies and in women with evidence of HPV31 infection. Neutralizing antibodies were detected in 78% of subjects with a positive enzyme-linked immunosorbent assay.

scholarly journals Enzyme-Linked Immunosorbent Assay To Differentiate the Antibody Responses of Animals Infected with Brucella Species from Those of Animals Infected with Yersinia enterocolitica O9

2003 ◽  
Vol 10 (4) ◽  
pp. 710-714 ◽  
Author(s):  
Janchivdorj Erdenebaatar ◽  
Balgan Bayarsaikhan ◽  
Masahisa Watarai ◽  
Sou-ichi Makino ◽  
Toshikazu Shirahata
Keyword(s):  
Yersinia Enterocolitica ◽  
Immunosorbent Assay ◽  
Brucella Abortus ◽  
Field Trial ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Brucella Species ◽  
Serological Tests ◽  
Human Sera ◽  
Immunosorbent Assays

ABSTRACT Enzyme-linked immunosorbent assays using antigens extracted from Brucella abortus with n-lauroylsarcosine differentiated natural Brucella-infected animals from Brucella-vaccinated or Yersinia enterocolitica O9-infected animals. A field trial in Mongolia showed cattle, sheep, goat, reindeer, camel, and human sera without infection could be distinguished from Brucella-infected animals by conventional serological tests.

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.

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