Canine Brugia malayi microfilarial excretory/secretory protein-based antibody assay for the diagnosis of brugian filariasis in dogs
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Brugian filariasis is reported in dogs in Kerala, India. Antibody detection kits are not available worldwide, for detection of Brugian filariasis in dogs. A study was carried out to develop Indirect plate ELISA using excretory secretory antigen isolated from canine brugian microfilariae and compare the sensitivity and specificity with that of blood smear examination. Identification of microfilariae was done by acid phosphatase staining using Naphthol AS-TR method and Polymerase Chain Reaction for Hha 1 repeat sequence. The microfilariae were identified as Brugia malayi. Isolation of brugian microfilariae from canine blood was done by gradient centrifugation method. The isolated microfilariae were maintained in RPMI-1640 media. The pooled media was then concentrated to obtain excretory secretory protein (ESP). This ESP was used to develop Indirect ELISA. The sensitivity and specificity of the plate ELISA developed was 84 and 100 per cent respectively when compared with blood smear examination. This is the first report of successful isolation of ESP from Brugia malayi microfilariae from dogs and standardization of plate ELISA using the antigen.
KeywordsBrugia malayi Dog Kerala Excretory secretory protein (ESP) Plate ELISA
The present paper is a part of MVSc thesis submitted to the Kerala Veterinary and Animal Sciences University and the authors are thankful to the University for the facilities and Grant rendered for the research.
PVS: MVSc scholar performed the research. DC: Major guide for the MVSc Research who formulated the research work and gave instructions in performing the research and helped in preparing the manuscript. UNP: Minor guide for the MVSc research and Head of the Department who provided all guidance for the research. NMU: Minor Guide for the MVSc Research and guided for the research. BL: Minor Guide for the MVSc Research and guided for ELISA standardization. CS: Helped in analyzing the data statistically.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
The project is formulated following the guidelines of Institutional Animal Ethics Committee and got approval from Faculty Research Committee with code number Ad/9/96/MVM/2015/CM. It is certified that all biomaterials required for the research were collected with prior written consent from the owners of the animals.
The article is a part of M.V.Sc. thesis submitted to Kerala Veterinary and Animal Sciences University and submitted with consent from all authors.
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