Journal of Parasitic Diseases

, Volume 43, Issue 4, pp 549–553 | Cite as

Canine Brugia malayi microfilarial excretory/secretory protein-based antibody assay for the diagnosis of brugian filariasis in dogs

  • Poojary Vineeta Sadarama
  • Deepa ChirayathEmail author
  • Usha Narayana Pillai
  • N. Madhavan Unny
  • Bindu Lakshmanan
  • C. Sunanda
Original Article


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.


Brugia 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.

Author contributions

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.

Ethical statement

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.

Informed consent

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.


  1. Ambily VR, Pillai UN, Arun S, Pramod S, Jayakumar KM (2011) Detection of human filarial parasite B. malayi in dogs by histochemical staining and molecular techniques. Vet Parasitol 181:210–214CrossRefGoogle Scholar
  2. Bomfim MRQ, Ko A, Koury MC (2005) Evaluation of the recombinant LipL32 in enzyme-linked immunosorbent assay for the serodiagnosis of bovine leptospirosis. Vet Mic 109(1):89–94CrossRefGoogle Scholar
  3. Chalifoux L, Hunt RD (1971) Histochemical differentiation of Dirofilaria immitis and Dipetalonema reconditum. J Am Vet Med Assoc 158(5):601–605PubMedGoogle Scholar
  4. Chenthamarakshan V, Reddy MVR, Harinath BC (1996) Diagnostic potential of fractionated Brugia malayi microfilarial excretory/secretory antigen for bancroftianfilariasis. Trans R Soc Trop Med Hyg 90(3):252–254CrossRefGoogle Scholar
  5. Chirayath D, Alex PC, Usha NP (2015) Microscopic characteristics of canine filariasis in Thrissur district of Kerala. Indian Vet J 92:81–82Google Scholar
  6. Chirayath D, Alex PC, Usha NP, George S, Ajithkumar S, Panicker VP (2017) Identification of Brugia malayi in dogs in Kerala, India. Trop Biomed 34:804–814Google Scholar
  7. Kumari S, Lillibridge CD, Bakeer M, Lowrie RC, Jayaraman K, Philipp MT (1994) Brugia malayi: the diagnostic potential of recombinant excretory/secretory antigens. Exp Parasitol 79(4):489–505CrossRefGoogle Scholar
  8. Marley SE, Lammie PJ, Eberhard ML, Hightower AW (1995) Reduced antifilarial IgG4 responsiveness in a subpopulation of microfilaremic persons. J Infect Dis 172(6):1630–1633CrossRefGoogle Scholar
  9. Noordin R, Shenoy RK, Rahman RA (2003) Comparison of two IgG4 assay formats (ELISA and rapid dipstick test) for detection of brugian filariasis. Southeast Asian J Trop Med Public Health 34(4):768–770PubMedGoogle Scholar
  10. Rahmah N, Khairul AA, Tengku ARH, Zurainee MN, Noor A’Shikin A, Fadzillah A, Maimunah A, Ashraful HJ (1998) Use of antifilarial IgG4-ELISA to detect Brugia malayi infection in an endemic area of Malaysia. Trop Med Int Health 3(3):184–188CrossRefGoogle Scholar
  11. Srivastava P, Bhattacharjee J, Dhariwal A, Krishnamoorthy K, Dash A (2014) Elimination of lymphatic filariasis-current status and way ahead. J Commun Dis 46(2):85–94Google Scholar
  12. Xie H, Bain O, Williams SA (1994) Molecular phylogenetic studies on Brugia filariae using Hha I repeat sequences. Parasite 1(3):255–260CrossRefGoogle Scholar

Copyright information

© Indian Society for Parasitology 2019

Authors and Affiliations

  • Poojary Vineeta Sadarama
    • 1
  • Deepa Chirayath
    • 1
    Email author
  • Usha Narayana Pillai
    • 1
  • N. Madhavan Unny
    • 1
  • Bindu Lakshmanan
    • 2
  • C. Sunanda
    • 3
  1. 1.Department of Veterinary Clinical Medicine, Ethics & Jurisprudence, College of Veterinary and Animal Sciences, MannuthyKerala Veterinary and Animal Sciences UniversityPookodeIndia
  2. 2.Department of Veterinary Parasitology, College of Veterinary and Animal Sciences, MannuthyKerala Veterinary and Animal Sciences UniversityPookodeIndia
  3. 3.Department of Statistics, College of Veterinary and Animal Sciences, PookodeKerala Veterinary and Animal Sciences UniversityPookodeIndia

Personalised recommendations