دورية أكاديمية
A PCR-ELISA for the identification of cyathostomin fourth-stage larvae from clinical cases of larval cyathostominosis.
العنوان: | A PCR-ELISA for the identification of cyathostomin fourth-stage larvae from clinical cases of larval cyathostominosis. |
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المؤلفون: | Hodgkinson JE; Department of Veterinary Parasitology, Liverpool School of Tropical Medicine, Pembroke Place, L3 5QA, Liverpool, UK. jhodgkin@liverpool.ac.uk, Lichtenfels JR, Mair TS, Cripps P, Freeman KL, Ramsey YH, Love S, Matthews JB |
المصدر: | International journal for parasitology [Int J Parasitol] 2003 Oct; Vol. 33 (12), pp. 1427-35. |
نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. |
اللغة: | English |
بيانات الدورية: | Publisher: Elsevier Science Country of Publication: England NLM ID: 0314024 Publication Model: Print Cited Medium: Print ISSN: 0020-7519 (Print) Linking ISSN: 00207519 NLM ISO Abbreviation: Int J Parasitol Subsets: MEDLINE |
أسماء مطبوعة: | Publication: Oxford : Elsevier Science Original Publication: Oxford, New York, Pergamon Press. |
مواضيع طبية MeSH: | Genes, Helminth*, Strongyle Infections, Equine/*diagnosis , Strongylida/*genetics, Animals ; Blotting, Southern/methods ; Enzyme-Linked Immunosorbent Assay/methods ; Female ; Horses ; Larva ; Male ; Oligonucleotide Probes ; Polymerase Chain Reaction/methods ; Reproducibility of Results |
مستخلص: | We report the use of six oligoprobes designed from intergenic spacer region sequences to identify fourth-stage larvae (L4) of the tribe Cyathostominae. Oligoprobes were designed for identification of the following species: Cylicocyclus ashworthi, Cylicocyclus nassatus, Cylicocyclus insigne, Cyathostomum catinatum, Cylicostephanus goldi, and Cylicostephanus longibursatus. A seventh probe was designed as a positive control to identify all these members of the Cyathostominae. The intergenic spacer region was amplified by PCR using conserved primers. Initially, three oligoprobes were used in Southern blot analysis. To facilitate high-throughput identification, these and a further four oligoprobes were developed for use in a PCR-ELISA. All probes were validated for their ability to detect cyathostomin PCR products in the PCR-ELISA, using DNA from morphologically identified adult parasites. Initially, 712 L4 were isolated from the diarrhoeic faeces from horses (n=17) with clinical larval cyathostominosis. PCR products from 522 of these L4 were subjected to analysis, with 413 L4 being identified as one of the aforementioned species. With reference to individual species analysis, 28.5% of the 522 L4 were identified as C. longibursatus, 25.7% as C. nassatus, 15.9% as C. ashworthi, 7.3% as C. goldi and 1.7% as C. catinatum. No L4 were identified as being C. insigne species. When L4 within faeces from individual horses were compared, no sample was found to comprise parasites of one species. The least number of species identified in a single sample was two. This study suggests that clinical larval cyathostominosis is predominantly caused by mixed-species infections. |
المشرفين على المادة: | 0 (Oligonucleotide Probes) |
تواريخ الأحداث: | Date Created: 20031007 Date Completed: 20040330 Latest Revision: 20190827 |
رمز التحديث: | 20240628 |
DOI: | 10.1016/s0020-7519(03)00140-1 |
PMID: | 14527525 |
قاعدة البيانات: | MEDLINE |
تدمد: | 0020-7519 |
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DOI: | 10.1016/s0020-7519(03)00140-1 |