Evaluation of PCR, IEF and ELISA techniques for the detection and identification of potato cyst nematodes from field soil samples in England and Wales

• Published in: Pest management science 2001-11, Vol.57 (11), p.1068-1074
• Main Authors: Ibrahim, Said K, Minnis, Stephen T, Barker, Anthony D P, Russell, Mike D, Haydock, Patrick P J, Evans, Kenneth, Grove, Ivan G, Woods, Simon R, Wilcox, Andrew
• Format: Article
• Language: English
• Subjects: Animals; bait test; Biological and medical sciences; cyst nematodes; DNA; Electrophoresis, Gel, Pulsed-Field - methods; ELISA; England; Enzyme-Linked Immunosorbent Assay - methods; Fundamental and applied biological sciences. Psychology; Globodera pallida; Globodera rostochiensis; IEF; Nematoda - genetics; Nematoda - growth & development; PCN; PCR; Phytopathology. Animal pests. Plant and forest protection; Plant Diseases - parasitology; Polymerase Chain Reaction - methods; Protozoa. Invertebrates; Records, symptoms, damages, economic importance, population surveys; Reproducibility of Results; rITS; Solanum tuberosum - parasitology; Statistics as Topic; Wales
• ISSN: 1526-498X; 1526-4998
• DOI: 10.1002/ps.397

Effective management of potato cyst nematodes (PCNs) requires simple, rapid and accurate identification and quantification of field populations. Soil samples from a survey of 484 fields in potato rotations in England and Wales were used to compare the identification and quantification of PCNs using IEF, PCR, ELISA and bait plant tests. The cyst counts and bait plant test revealed that 64.3% of field samples contained PCNs. Bait plant tests increased the detection rate of PCNs in field samples by 4–6.4%. This means that some infestations are cryptic and would not normally be detected by standard counts. IEF, PCR and ELISA methods distinguished between Globodera rostochiensis and G pallida and were able to register mixed populations; however they were not in full agreement. All methods suggested that G pallida is the dominant species in the field samples tested. The PCR results indicated that 66% of field samples contained pure G pallida, 8% contained pure G rostochiensis and 26% contained mixtures of the two species. Estimates of the relative process times taken per sample in the PCR, IEF and ELISA techniques are given. © 2001 Society of Chemical Industry