[Detection of Brugia malayi larva in mosquito vector by PCR and PCR-ELISA].
OBJECTIVE
To develop sensitive, specific, simple assays for the detection of Brugia malayi larva in mosquito vectors.


METHODS
The optimum conditions for PCR and PCR-ELISA were determined. With dissected larvae, detection limits and as well as specificities of the two assays were worked out. The optimized assays were then tested with infected and non-infected mosquito vectors, Anopheles sinensis.


RESULTS
Both PCR and PCR-ELISA detected DNA equivalent to a single L1-stage larva(L1). While the actual detecting limits of the two, as estimated and titrated respectively, reached to 1/10 and 1/100 of a L1. When the two assays were used to detect the larvae isolated from 113 infected A. sinensis, all gave specific bands and positive reactions. However, trials on direct amplifications with crude extracts of infected mosquito vectors consistently failed to give specific bands upon electrophoresis and negative results in PCR-ELISA as well.


CONCLUSIONS
Both PCR and PCR-ELISA were preliminarily established for the detection of isolated B. malayi larva in mosquito vector, which proved to be sensitive, specific and easy to manipulate.
Liu
Y
Y
Institute of Parasitic Diseases, Chinese Academy of Preventive Medicine, Shanghai 200025.
Sun
D
D
Xue
H
H
D000818:Animals; D000852:Anopheles / Q000469:parasitology*; D017178:Brugia malayi / Q000302:isolation & purification*; D018509:DNA, Helminth / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D007303:Insect Vectors / Q000469:parasitology*; D007814:Larva; D016133:Polymerase Chain Reaction
D018509:DNA, Helminth