In search of a potential diagnostic tool for molecular characterization of lymphatic filariasis

Mohd Saeed 1 , 2 , Mohd Adnan 3 , Saif Khan 3 , Eyad Al-Shammari 3 , and Huma Mustafa 4
  • 1 Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, University of Ha’il, Ha’il, PO Box 2440, Kingdom of Saudi Arabia
  • 2 Department of Biosciences, Integral University, Lucknow, Uttar Pradesh, 226026, India
  • 3 Department of Clinical Nutrition, College of Applied Medical Sciences, University of Ha’il, Ha’il, PO Box 2440, Kingdom of Saudi Arabia
  • 4 Council of Science and Technology, Vigyan Bhawan, Lucknow, 226018, Uttar Pradesh, India


Lymphatic filariasis (LF) is a chronic disease and is caused by the parasites Wuchereria bancrofti (W. bancrofti), Brugia malayi (B. malayi) and Brugia timori (B. timori). In the present study, Setaria cervi (S. cervi), a bovine filarial parasite has been used. Previously, it has been reported that the S. cervi shares some common proteins and antigenic determinants with that of human filarial parasite. The larval stages of filarial species usually cannot be identified by classical morphology. Hence, molecular characterization allows the identification of the parasites throughout all their developmental stages. The genomic DNA of S. cervi adult were isolated and estimated spectrophotometrically for the quantitative presence of DNA content. Screening of DNA sequences from filarial DNA GenBank and Expressed Sequence Tags (EST’s) were performed for homologous sequences and then multiple sequence alignment was executed. The conserved sequences from multiple sequence alignment were used for In Silico primer designing. The successfully designed primers were used further in PCR amplifications. Therefore, in search of a promising diagnostic tool few genes were identified to be conserved in the human and bovine filariasis and these novel primers deigned may help to develop a promising diagnostic tool for identification of lymphatic filariasis.

  • Alasaad S., Pascucci I., Jowers M.J., Soriguer R.C., Zhu X.Q., Rossi L. 2012. Phylogenetic study of Setaria cervi based on mitochondrial cox1 gene sequences. Parasitology Research, 110, 281-285

  • Bockarie M.J., Molyneux D.H. 2009. The end of lymphatic filariasis? British Medical Journal (BMJ), 338, 1686

  • Braga C., Dourado M.I., Ximenes R.A.D.A., Alves L., Brayner F., Rocha A., Alexander N. 2003. Field evaluation of the whole blood immunochromatographic test for rapid bancroftian filariasis diagnosis in the northeast of Brazil. Revista do Instituto de Medicina Tropical de São Paulo, 45, 125-129

  • Cancrini G., Kramer L.H. 2001. Insect vectors of Dirofilaria spp. In: Simon F., Genchi C. (eds) Heartworm infection in humans and animals. Universidad de Salamanca, Spain, 63-82

  • Chu B.K., Hooper P.J., Bradley M.H., McFarland D.A., and Ottesen E.A. 2010.The economic benefits resulting from the first 8 years of the Global Programme to Eliminate Lymphatic Filariasis (2000-2007). PLoS Neglected Tropical Diseases, 4, 708

  • Cooper R.D. 1998 Preservation of anopheline mosquitoes for DNA probe analysis. Joumal of the American Mosquito Control Association, 14, 58-60

  • Dissanayake S., Piessens W.F. 1991. Detection of amplified Wuchereria bancrofti DNA in mosquitoes with a non-radioactive probe. Molecular and Biochemical Parasitology, 45, 49-56

  • Dixit V, Pati AK, Gupta AK, Bisen PS, Prasad GB. 2009. Filarial infection is resisted differentially by subjects having different blood group phenotypes. Journal of Clinical Laboratory Analysis, 23, 86-91. DOI: 10.1002/jcla.20317

  • Farid H.A., Hammad R.E., Hassan M.M., Morsy Z.S., Kamal I.H., Weil G.J., Ramzy R.M.R. 2001. Detection of Wuchereria bancrofti in mosquitoes by the polymerase chain reaction: a potentially useful tool for large scale control programmes. Transactions of the Royal Society of Tropical Medicine and Hygine, 95, 29-32

  • Favia G., Lanfrancotti A., Della Torre A., Cancrini G., Coluzzi M. 1997. Advances in the identification of Dirofilaria repens and Dirofilaria immitis by a PCR-based approach. Parassitologia, 39, 401-402

  • Ghedin E., Wang S., Spiro D., Caler E., Zhao Q., Crabtree J., Allen J.E., Delcher A.L., Guiliano D.B., Miranda-Saavedra D., Angiuoli S.V., Creasy T., Amedeo P., Haas B., El-Sayed N.M., Wortman J.R., Feldblyum T., Tallon L., Schatz M., Shumway M., Koo H., Salzberg S.L., Schobel S., Pertea M., Pop M., White O., Barton G.J., Carlow C.K., Crawford M.J., Daub J., Dimmic M.W., Estes C.F., Foster J.M., Ganatra M., Gregory W.F., Johnson N.M., Jin J., Komuniecki R., Korf I., Kumar S., Laney S., Li B.W., Li W., Lindblom T.H., Lustigman S., Ma D., Maina C.V., Martin D.M., McCarter J.P., McReynolds L., Mitreva M., Nutman T.B., Parkinson J., Peregrín-Alvarez J.M., Poole C., Ren Q., Saunders L., Sluder A.E., Smith K., Stanke M., Unnasch T.R., Ware J., Wei A.D., Weil G., Williams D.J., Zhang Y., Williams S.A., Fraser-Liggett C., Slatko B., Blaxter M.L., and Scott A.L. 2007. Draft genome of the filarial nematode parasite Brugia malayi. Science, 317, 1756-1760

  • Hamlin K.L., Moss D.M., Priest J.W., Roberts J., Kubofcik J., Gass K., Lammie P. J. 2012. Longitudinal monitoring of the development of antifilarial antibodies and acquisition of Wuchereria bancrofti in a highly endemic area of Haiti. PLoS Neglected Tropical Diseases, 6, e1941

  • Hedge E.C., Ridley D.S. 1977. Immunofluorescent reactions with microfilariae: 1. Diagnostic evaluation. Transactions of the Royal Society of Tropical Medicine and Hygine 71, 304

  • Kaushal N.A., Hussain R., Nash E., Ottesen E.A. 1982. Identification and characterization of excretory-secretory products of Brugia malayi, adult filarial parasites. The Journal of Immunology, 129, 338-343

  • Kolaczinski J. H., Onapa A. W., Kabatereine N. B., Ndyomugyenyi R., Kakembo A. S., Brooker S. 2006. Neglected tropical diseases and their control in uganda. Analysis, chapter 4, pp. 27-31

  • Koroma J. B., Bangura M. M., Hodges M. H., Bah M. S., Zhang Y., Bockarie M. J. 2012. Lymphatic filariasis mapping by Immunochromatographic Test cards and baseline microfilaria survey prior to mass drug administration in Sierra Leone. Parasites & Vectors, 5, 10 DOI: 10.1186/1756-3305-5-10

  • Krushna N., Shiny C. 2009 Immunolocalization and serum antibody responses to Brugia malayi pepsin inhibitor homolog (B-33). Microbiology and Immunology, 53, 173-183

  • Kubofcik J., Fink D.L., Nutman T.B. 2012. Identification of Wb123 as an Early and Specific Marker of Wuchereria bancrofti Infection. PLoS Neglected Tropical Diseases, 6, e1930. DOI:10.1371/journal.pntd.0001930

  • Lizotte M.R., Supali T., Partono F. and Williams S.A. 1994. A polymerase chain reaction assay for the detection of Brugia malayi in blood. American Journal Tropical Medicine and Hygiene, 51, 314-21

  • Madathiparambil M.G., Kaleysa K.N., Raghavan K. 2009. A diagnostically useful 200-kDa protein is secreted through the surface pores of the filarial parasite Setaria digitata. Parasitology Research, 105, 1099-1104

  • McCarthy J.S., Lustigman S., Yang G.J., Barakat R.M., Garcia H.H., Sripa B., Willingham A.L., Prichard R.K., Basáñez M.G. 2012.A research agenda for helminth diseases of humans: diagnostics for control and elimination programmes. PLoS Neglected Tropical Diseases, 6, 1601. DOI: 10.1371/journal. pntd.0001601

  • McCarthy J.S., Zhong M., Gopinath R. (1996). Evaluation of a polymerase chain reaction-based assay for diagnosis ofWuchereria bancrofti infection. Journal of Infectious Disease, 173, 1510-4

  • McReynolds L.A., DeSimone S.M., Williams S.A. 1986. Cloning and comparison of repeated DNA sequences from the human filarial parasite Brugia malayi and the animal parasite Brugia pahangi. Proceedings of the National Academic of Science, USA 83, 797-801

  • Melrose W. 2004. Lymphatic filariasis: A review 1862-2002 (pp. 1-80). Warwick Educational Publishing

  • Molyneux D.H., Taylor M.J. 2001 Current status and future prospects of the Global Lymphatic Filariasis Programme. Current Opinion in Infectious Diseases, 14, 155-159

  • Mustafa H., Srivastava N., Kaushal D.C., Kaushal, N. A. 1996. Analysis and potential of excretory-secretory antigens of Setaria cervi for immunodiagnosis of human filariasis. Indian Journal of Experimental Biology, 34, 508-512

  • Nanduri J., Kazura J.W. (1989). Clinical and laboratory aspects of filariasis. Clinical Microbiology Reviews, 2, 39-50

  • Paily K.P., Hoti S.L., Das P.K. 2009. A review of the complexity of biology of lymphatic filarial parasites. Journal of Parasitic Diseases, 33, 3-12

  • Pandiaraja P., Arunkumar C., Hoti S. 2010. Evaluation of synthetic peptides of WbSXP-1 for the diagnosis of human lymphatic filariasis. Diagnostic Microbiology and Infectious Disease, 68, 410-415

  • Poole C.B., William, S.A. 1990. A rapid DNA assay for the speciesspecific detection and quantification of Brugia in blood sample. Molecular and Biochemical Parasitiology, 40, 129-36

  • Praphathip E., Phaik-Eem L., Hoi S.Y. 2013. The raffles bulletin of zoology 2013 Supplement No.29: 99-109

  • Rahman A.R., Hwen-Yee C., Noordin R. 2007. Pan LF-ELISA using BmR1 and BmSXP recombinant antigens for detection of lymphatic filariasis. Filaria Journal, 6, 10. DOI:10.1186/ 1475-2883-6-10

  • Siridewa K., Karunanayake E.H., Chandrasekharan N.V., Abeyewickreme W., Franzen L., Aslund L., Pettersson U. 1994. Cloning and characterization of a repetitive DNA sequence specific for Wuchereria bancrofti. The American Journal of Tropical Medicine and Hygiene, 51, 495-500

  • Srinivasan L., Mathew N., Muthuswamy K. 2009. In vitro antifilarial activity of glutathione S-transferase inhibitors. Parasitology Research, 105, 1179-1182

  • Steel C., Kubofcik J., Ottesen E.A., Nutman T.B. 2012. Antibody to the Filarial Antigen Wb123 Reflects Reduced Transmission and Decreased Exposure in Children Born following Single Mass Drug Administration (MDA). PLoS Neglected Tropical Diseases, 6, e1940. DOI:10.1371/journal.pntd.0001940

  • Thompson J.D., Gibson T.J., Higgins D.G. Multiple sequence alignment using ClustalW and Clustal X. Current Protocols in Bioinformatics. 2002; Chapter 2 (Unit 2 3).(chapter 4)

  • Walther M., Muller R. 2003. Diagnosis of human filariases (except onchocerciasis). Advances in parasitology, 53, 149-193

  • Weil G., Curtis K., Fischer P., Won K. 2011.A multicenter evaluation of a new antibody test kit for lymphatic filariasis employing recombinant Brugia malayi antigen Bm-14. Acta Tropica, 120, 1-9. DOI:10.1016/j.actatropica.2010.04.010.A

  • WHO. 2014 Lymphatic filariasis Fact sheet Updated March 2014. (

  • WHO. 2005. Sixth meeting of the Technical Advisory Group on the Global Elimination of Lymphatic Filariasis, Geneva, Switzerland, 20-23 September 2005. The Weekly Epidemiological Record, 80, 401-408

  • Williams S.A., DeSimone S.M., McReynolds L.A. 1988. Speciesspecific oligonucleotide probes for the identification of human filarial parasites. Molecular and Biochemical Parasitology, 28, 163-169

  • Zhong, M., McCarthy, J.S., Bierwert, L., Lizotte-Waniewski M., Chanteau, S., Nutman T.B., Ottesen E. and Williams S.A. (1996). A polymerase chain reaction assay for detection of the parasite Wuchereria bancrofti in human blood samples. The American Journal of Tropical Medicine and Hygiene, 54, 357-63

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