Serologically Based Diagnostic and Quantification Tests for Nematodes

Davies, Keith; Curtis, R. H. C.; Evans, K.

doi:10.1002/(sici)1096-9063(199605)47:1<81::aid-ps388>3.0.co;2-2

Cited by 10 publications

(4 citation statements)

References 10 publications

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“…Antibodies, both polyclonal and monoclonal, have been produced for Meloidogyne spp. diagnostics, but the use of antibodies is limited to a few examples as it has, according to Blok & Powers (2009), been superseded by DNA-based diagnostics (Davies et al, 1996;Tastet et al, 2001). In recent years, identification methods that are DNA-based have gained popularity and this has led to a combination of both morphological, biochemical and molecular methods to describe and identify nematodes (Karssen et al, 2004;Castillo et al, 2009).…”

Section: Species Identificationmentioning

confidence: 99%

The threat of root‐knot nematodes (Meloidogyne spp.) in Africa: a review

et al. 2014

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Meloidogyne species pose a significant threat to crop production in Africa due to the losses they cause in a wide range of agricultural crops. The direct and indirect damage caused by various Meloidogyne species results in delayed maturity, toppling, reduced yields and quality of crop produce, high costs of production and therefore loss of income. In addition, emergence of resistance-breaking Meloidogyne species has partly rendered various pest management programmes already in place ineffective, therefore putting food security of the continent at risk. It is likely that more losses may be experienced in the future due to the on-going withdrawal of nematicides. To adequately address the threat of Meloidogyne species in Africa, an accurate assessment and understanding of the species present, genetic diversity, population structure, parasitism mechanisms and how each of these factors contribute to the overall threat posed by Meloidogyne species is important. Thus, the ability to accurately characterize and identify Meloidogyne species is crucial if the threat of Meloidogyne species to crop production in Africa is to be effectively tackled. This review discusses the use of traditional versus molecular-based identification methods of Meloidogyne species and how accurate identification using a polyphasic approach can negate the eminent threat of root knot nematodes in crop production. The potential threat to Africa posed by highly damaging and resistance-breaking populations of 'emerging' Meloidogyne species is also examined.

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Section: Species Identificationmentioning

confidence: 99%

The threat of root‐knot nematodes (Meloidogyne spp.) in Africa: a review

et al. 2014

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“…However, only partial discrimination between adult females of the nematode species could be made. Davies et al (1996) and Lima et al (2005) raised monoclonal antibodies against M.incognita with an aim to identify the species, but they cross reacted with some other species of root knot nematodes. Hence, in the present studies, an attempt has been made to develop ELISA for specific identification of M.incognita.…”

Section: Discussionmentioning

confidence: 99%

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

2013

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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.

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“…G. rostochiensis and G. pallida can be distinguished by morphological features of the cysts but this requires a high degree of skill and is time consuming. A method based on iso‐electric focusing (IEF) of proteins extracted from cysts (Fleming and Marks, 1982) has been used as a diagnostic test, as have immunological methods (Davies et al ., 1996). More recently, PCR‐based test methods have become the method of choice for species identification (Mulholland et al ., 1996; Bulman and Marshall, 1997).…”

Section: Introductionmentioning

confidence: 99%

Determination of relative proportions of Globodera species in mixed populations of potato cyst nematodes using PCR product melting peak analysis

Bates

Taylor

Gans

et al. 2002

Molecular Plant Pathology

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Summary Using a novel approach based on melting peak analysis of PCR products, we have developed a semi-quantitative assay to measure the relative proportions of Globodera pallida and Globodera rostochiensis in a sample of potato cyst nematodes (PCN). The method depends on a competitive multiplex PCR where the products of each species can be separated by their distinct melting temperatures (T(m)). The melting curves of the products are measured by continual fluorescence monitoring in the presence of the intercalating dye SYBR Green I whilst gradually increasing the temperature. Varying the proportion of cysts of each PCN species gave rise to melting curves with different peak heights, which reflected the relative amounts of each DNA in the sample. By calculating the ratio of the melting peak heights at the Tm of each product and comparing it with standards run under the same conditions, it was possible to estimate the proportion of each product in the mixture. Sensitivity was such that 2% of G. pallida cysts in a mixture could be detected. All results predicted by the analysis were confirmed by agarose gel electrophoresis of the PCR products. The method is rapid, reproducible and following DNA extraction, up to 25 samples can be analysed in an hour.

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Serologically Based Diagnostic and Quantification Tests for Nematodes

Cited by 10 publications

References 10 publications

The threat of root‐knot nematodes (Meloidogyne spp.) in Africa: a review

The threat of root‐knot nematodes (Meloidogyne spp.) in Africa: a review

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

Determination of relative proportions of Globodera species in mixed populations of potato cyst nematodes using PCR product melting peak analysis

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