Application of local products R-phycoerythrin and monoclonal antibody as a fluorescent antibody probe to detect Entamoeba histolytica trophozoites.
Southeast Asian J Trop Med Public Health
;
1996 Jun; 27(2): 297-303
Article
in English
| IMSEAR
| ID: sea-35681
ABSTRACT
R-phycoerythrin (R-PE) was extracted from red algae, Gracilaria fisheri from Pattani Province, Thailand, with 50 mM sodium phosphate buffer, pH 7.0, followed by precipitation with 30-50% final concentration of saturated ammonium sulphate solution at 0 degree C. The precipitate was further purified by DEAE-cellulose (DE-52) column chromatography. The purified R-PE showed a single band of Mr 240 kDa by polyacrylamide gel electrophoresis with the maximum absorption and maximum fluorescence emission at 565 nm and at 573 nm respectively, and the OD ratio of 565 to 280 nm was 6.7. The IgG fraction of a murine monoclonal antibody (Eh208C2-2 MIgG) raised against trophozoites of HM-1 IMSS strain of pathogenic E. histolytica was conjugated with purified R-PE by using the heterobifunctional compound N-succinimidyl3-(2-pyridyldithio)propionate (SPDP). The conjugate was shown by direct immunofluorescent antibody (DIFA) assay to stain specifically both the culture-derived and stool-derived E. histolytica trophozoites.
Full text:
Available
Index:
IMSEAR (South-East Asia)
Main subject:
Phycoerythrin
/
Thailand
/
Humans
/
Immunoglobulin G
/
Antibodies, Protozoan
/
Reproducibility of Results
/
Sensitivity and Specificity
/
Fluorescent Antibody Technique, Direct
/
Dysentery, Amebic
/
Entamoeba histolytica
Type of study:
Diagnostic study
Country/Region as subject:
Asia
Language:
English
Journal:
Southeast Asian J Trop Med Public Health
Year:
1996
Type:
Article
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