Skip to main content
Top
Published in: Journal of Parasitic Diseases 3/2014

01-09-2014 | Original Article

Development of enzyme linked immunosorbent assay (ELISA) for the detection of root-knot nematode Meloidogyne incognita

Authors: J. Kapur-Ghai, M. Kaur, P. Goel

Published in: Journal of Parasitic Diseases | Issue 3/2014

Login to get access

Abstract

Root-knot nematodes (Meloidogyne incognita) are obligate, sedentary plant endoparasites that are extremely polyphagous in nature and cause severe economic losses in agriculture. Hence, it is essential to control the parasite at an early stage. For any control strategy to be effective, an early and accurate diagnosis is of paramount importance. Immunoassays have the inherent advantages of sensitivity and specificity; have the potential to identify and quantify these plant-parasitic nematodes. Hence, in the present studies, enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of M.incognita antigens. First an indirect ELISA was developed for detection and titration of anti-M.incognita antibodies. Results indicated as high as 320 K titre of the antisera. Finally competitive inhibition ELISA was developed employing these anti-M.incognita antibodies for detection of M.incognita antigens. Sensitivity of ELISA was 10 fg. Competitive inhibition ELISA developed in the present studies has the potential of being used as an easy, rapid, specific and sensitive diagnostic tool for the detection of M.incognita infection.
Literature
go back to reference Atkinson HJ, Harris PD, Halk EJ, Novitski C, Leighton-Sands J, Nolan P, Fox PC (1988) Monoclonal antibodies to the soya bean cyst nematode, Heterodera glycines. Ann Appl Biol 112:459–469CrossRef Atkinson HJ, Harris PD, Halk EJ, Novitski C, Leighton-Sands J, Nolan P, Fox PC (1988) Monoclonal antibodies to the soya bean cyst nematode, Heterodera glycines. Ann Appl Biol 112:459–469CrossRef
go back to reference Bellafiore S, Shen Z, Rosso MN, Abad P, Shih P, Briggs SP (2008) Direct identification of the Meloidogyne incognita secretome reveals proteins with host cell reprogramming potential. PLoS Pathog 4:e1000192PubMedCentralPubMedCrossRef Bellafiore S, Shen Z, Rosso MN, Abad P, Shih P, Briggs SP (2008) Direct identification of the Meloidogyne incognita secretome reveals proteins with host cell reprogramming potential. PLoS Pathog 4:e1000192PubMedCentralPubMedCrossRef
go back to reference Blok VC, Powers CO (2009) Biochemical and Molecular identification. In: Perry RN, Moens M, Starr JL (eds) Root-knot Nematodes. CABI, Wallingford, pp 98–118CrossRef Blok VC, Powers CO (2009) Biochemical and Molecular identification. In: Perry RN, Moens M, Starr JL (eds) Root-knot Nematodes. CABI, Wallingford, pp 98–118CrossRef
go back to reference Calvo E, Flores-Romero P, Lopez JA, Navas A (2005) Identification of proteins expressing differences among isolates of Meloidogyne incognita. (Nematoda: meloidogynidae) by nano-liquid chromatography coupled to ion-trap mass spectrometry. J Proteome Res 4:1017–1021PubMedCrossRef Calvo E, Flores-Romero P, Lopez JA, Navas A (2005) Identification of proteins expressing differences among isolates of Meloidogyne incognita. (Nematoda: meloidogynidae) by nano-liquid chromatography coupled to ion-trap mass spectrometry. J Proteome Res 4:1017–1021PubMedCrossRef
go back to reference Carneiro RMDG, Almeida MRA, Carneiro RG (1996) Enzyme phenotypes of Brazilian populations of Meloidogyne incognita. Fund Appl Nematol 19:555–560 Carneiro RMDG, Almeida MRA, Carneiro RG (1996) Enzyme phenotypes of Brazilian populations of Meloidogyne incognita. Fund Appl Nematol 19:555–560
go back to reference Chitwood DJ (2003) Research on plant-parasitic nematode biology conducted by the United States Department of Agriculture-Agricultural Research Service. Pest Mgmt Sci 59:748–753CrossRef Chitwood DJ (2003) Research on plant-parasitic nematode biology conducted by the United States Department of Agriculture-Agricultural Research Service. Pest Mgmt Sci 59:748–753CrossRef
go back to reference Curtis RHC, Al-Hinai MS, Diggines AER, Evans K (1997) Serological identification and quantification of Heterodera avenae from processed soil samples. Nematologica 43:199–213CrossRef Curtis RHC, Al-Hinai MS, Diggines AER, Evans K (1997) Serological identification and quantification of Heterodera avenae from processed soil samples. Nematologica 43:199–213CrossRef
go back to reference Davies KG, Lander EB (1992) Immunological differentiation of root-knot nematodes (Meloidogyne spp.) using monoclonal and polyclonal antibodies. Nematologica 38:353–366CrossRef Davies KG, Lander EB (1992) Immunological differentiation of root-knot nematodes (Meloidogyne spp.) using monoclonal and polyclonal antibodies. Nematologica 38:353–366CrossRef
go back to reference Davies KG, Curtis RH, Evans K (1996) Serologically based diagnostic and quantification tests for nematodes. Pestic Sci 47:81–87CrossRef Davies KG, Curtis RH, Evans K (1996) Serologically based diagnostic and quantification tests for nematodes. Pestic Sci 47:81–87CrossRef
go back to reference de Boer JM, Overmars HA, Pomp R, Davis EL, Zilverentant JF, Goverse A, Stokkermans JPWG, Hussey RH, Gommers FJ, Bakker J, Schots A (1996) Production and characterization of monoclonal antibodies to antigens from second-stage juveniles of the potato cyst-nematode Globodera rostochiensis. Fundl Appl Nematol 19:545–554 de Boer JM, Overmars HA, Pomp R, Davis EL, Zilverentant JF, Goverse A, Stokkermans JPWG, Hussey RH, Gommers FJ, Bakker J, Schots A (1996) Production and characterization of monoclonal antibodies to antigens from second-stage juveniles of the potato cyst-nematode Globodera rostochiensis. Fundl Appl Nematol 19:545–554
go back to reference El-Hady MM (2009) Discrimination among Meloidogyne incognita isolates by cellulase activity, protein and DNA fingprinting. Aust J Basic Appl Sci 3:1552–1558 El-Hady MM (2009) Discrimination among Meloidogyne incognita isolates by cellulase activity, protein and DNA fingprinting. Aust J Basic Appl Sci 3:1552–1558
go back to reference Engvall E, Perlmnan P (1971) Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry. 8:871–874 Engvall E, Perlmnan P (1971) Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry. 8:871–874
go back to reference Handoo ZA, Skantar AM, Carta LK, Erbe EF (2005) Morphological and molecular characterization of a new root-knot nematode, Meloidogyne thailandica n.sp. (Nematoda: meloidogynidae), parasitizing ginger (Zingiber sp.). J Nematol 37:343–353PubMedCentralPubMed Handoo ZA, Skantar AM, Carta LK, Erbe EF (2005) Morphological and molecular characterization of a new root-knot nematode, Meloidogyne thailandica n.sp. (Nematoda: meloidogynidae), parasitizing ginger (Zingiber sp.). J Nematol 37:343–353PubMedCentralPubMed
go back to reference Kennedy MJ, Schoelz JE, Donald PA, Niblack TL (1997) Unique immunogenic proteins in Heterodera glycines egg shells. J Nematol 29:276–281PubMedCentralPubMed Kennedy MJ, Schoelz JE, Donald PA, Niblack TL (1997) Unique immunogenic proteins in Heterodera glycines egg shells. J Nematol 29:276–281PubMedCentralPubMed
go back to reference Lima LM, Grossi-se-sa MF, Curtis RHC (2005) Immunolocalisation of secreted-excreted products of Meloidogyne spp. using polyclonal and monoclonal antibodies. Fitopatol Bras 30:629–633CrossRef Lima LM, Grossi-se-sa MF, Curtis RHC (2005) Immunolocalisation of secreted-excreted products of Meloidogyne spp. using polyclonal and monoclonal antibodies. Fitopatol Bras 30:629–633CrossRef
go back to reference Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J BiolChem 193:265–275 Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J BiolChem 193:265–275
go back to reference Palmer HM, Atkinson HJ and Perry RN (1992) Monoclonal antibodies (Mabs) specific to surface expressed antigens of Ditylenchus dipsaci. Fundam Appl Nematol 15:511–515 Palmer HM, Atkinson HJ and Perry RN (1992) Monoclonal antibodies (Mabs) specific to surface expressed antigens of Ditylenchus dipsaci. Fundam Appl Nematol 15:511–515
go back to reference Qiu JJ, Westerdahl BB, Anderson C, Williamson VM (2006) Sensitive PCR detection of Meloidogyne arenaria, M.incognita, and M javanica extracted from soil. J Nematol 38:434–441PubMedCentralPubMed Qiu JJ, Westerdahl BB, Anderson C, Williamson VM (2006) Sensitive PCR detection of Meloidogyne arenaria, M.incognita, and M javanica extracted from soil. J Nematol 38:434–441PubMedCentralPubMed
go back to reference Trudgill DL, Blok VC (2001) Apomictic, polyphagous root-knot nematodes: exceptionally successful and damaging biotrophic root pathogens. Annu Rev Phytopathol 39:53–77PubMedCrossRef Trudgill DL, Blok VC (2001) Apomictic, polyphagous root-knot nematodes: exceptionally successful and damaging biotrophic root pathogens. Annu Rev Phytopathol 39:53–77PubMedCrossRef
go back to reference Xu J, Liu P, Meng Q, Long H (2004) Characterisation of Meloidogyne species from China using isozyme phenotypes and amplified mitochondrial DNA restriction fragment length polymorphism. Eur J Plant Pathol 110:309–315CrossRef Xu J, Liu P, Meng Q, Long H (2004) Characterisation of Meloidogyne species from China using isozyme phenotypes and amplified mitochondrial DNA restriction fragment length polymorphism. Eur J Plant Pathol 110:309–315CrossRef
Metadata
Title
Development of enzyme linked immunosorbent assay (ELISA) for the detection of root-knot nematode Meloidogyne incognita
Authors
J. Kapur-Ghai
M. Kaur
P. Goel
Publication date
01-09-2014
Publisher
Springer India
Published in
Journal of Parasitic Diseases / Issue 3/2014
Print ISSN: 0971-7196
Electronic ISSN: 0975-0703
DOI
https://doi.org/10.1007/s12639-013-0246-0

Other articles of this Issue 3/2014

Journal of Parasitic Diseases 3/2014 Go to the issue
Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.