Detection of Paragonimus heterotremus in experimentally infected cat feces by antigen capture-ELISA and by DNA hybridization.
An antigen capture enzyme-linked immunosorbent assay (antigen capture-ELISA) and DNA hybridization technique were developed and evaluated for their application in the detection of Paragonimus heterotremus infection in experimentally infected cats. An IgG fraction prepared from serum of a rabbit immunized with P. heterotremus excretory-secretory (ES) products was used as the capture antibody. An IgG1 monoclonal antibody specific to the 22- and 31.5-kDa ES products of P. heterotremus was used as the antigen probe. As little as 0.24 ng of the ES products could be detected by this technique. A specific P. heterotremus DNA probe derived from the P. heterotremus genomic DNA library containing 1,500 base pairs was used in a dot-blot hybridization assay for the detection of parasite DNA. The radioactively labeled probe could detect DNA released from as few as 2 P. heterotremus eggs. Both ELISA and DNA hybridization were found to have 100% specificity, with sensitivities of 73.7% and 100%, respectively.
Maleewong
W
W
Department of Parasitology, Faculty of Medicine, Khon Kaen University, Thailand.
Intapan
P M
PM
Wongkham
C
C
Wongratanacheewin
S
S
Tapchaisri
P
P
Morakote
N
N
Chaicumpa
W
W
D000818:Animals; D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth / Q000032:analysis* / Q000276:immunology / Q000502:physiology; D002415:Cats; D018509:DNA, Helminth / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005243:Feces / Q000737:chemistry / Q000469:parasitology*; D005260:Female; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009693:Nucleic Acid Hybridization; D010237:Paragonimiasis / Q000175:diagnosis / Q000469:parasitology*; D010238:Paragonimus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D011817:Rabbits; D012680:Sensitivity and Specificity
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D018509:DNA, Helminth