A multiplex PCR assay for the simultaneous detection and discrimination of the sevenEimeriaspecies that infect domestic fowl
| Autor: | A H Pagotto, M M Furtado, Sandra Fernandez, Angela M. Katsuyama, Arthur Gruber, Alda Maria Backx Noronha Madeira |
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| Rok vydání: | 2003 |
| Předmět: |
Genetic Markers
Fowl Polymerase Chain Reaction Sensitivity and Specificity Eimeria law.invention Apicomplexa law Multiplex polymerase chain reaction Animals Multiplex Poultry Diseases Polymerase chain reaction biology Coccidiosis Genetic Variation Reproducibility of Results DNA Protozoan biology.organism_classification Molecular biology Random Amplified Polymorphic DNA Technique Infectious Diseases Genetic marker Animal Science and Zoology Parasitology Chickens Specific identification |
| Zdroj: | Parasitology. 127:317-325 |
| ISSN: | 1469-8161 0031-1820 |
| DOI: | 10.1017/s0031182003003883 |
| Popis: | This study reports the development of a novel multiplex PCR assay based on SCAR (Sequence-Characterised Amplified Region) markers for the simultaneous diagnosis of the 7Eimeriaspecies that infect domestic fowl. Primer pairs specific for each species were designed in order to generate a ladder of amplification products ranging from 200 to 811 bp. Sensitivity tests for each species were carried out, showing a detection threshold of 1–5 pg, which corresponds approximately to 2–8 sporulated oocysts. Distinct isolates of the 7Eimeriaspecies from different geographical sources were tested and successfully detected by the assay. All the species were amplified homogeneously, whether or not one of them was present in a high quantity, indicating that there was no cross-interference. The assay was also tested with different sources ofTaqDNA polymerase and thermocycler models, confirming the high reproducibility of the reaction. The economy of consumables and labour represented by a single-tube reaction greatly facilitates the molecular diagnosis of a large number of samples, making it appropriate for field epizootiological surveys. We propose the use of this multiplex PCR assay as a rapid and cost-effective diagnostic method for the detection and discrimination of the 7Eimeriaspecies that infect domestic fowl. |
| Databáze: | OpenAIRE |
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