Development of a nested polymerase chain reaction assay for the detection of Eimeria macusaniensis in camelid feces.

TitleDevelopment of a nested polymerase chain reaction assay for the detection of Eimeria macusaniensis in camelid feces.
Publication TypeJournal Article
Year of Publication2012
AuthorsCebra CK, Stang BV, Smith CC
JournalAmerican journal of veterinary research
Volume73
Issue1
Pagination13-8
Date Published2012 Jan
ISSN1943-5681
KeywordsAnimals, Base Sequence, Camelids, New World, Coccidiosis, DNA, Intergenic, DNA, Protozoan, Eimeria, Feces, Female, Male, Molecular Sequence Data, Oocysts, Oregon, Polymerase Chain Reaction
Abstract

OBJECTIVE: To test the ability of a nested PCR assay to detect Eimeria macusaniensis at various stages of infection in alpacas.

ANIMALS: 4 healthy adult alpacas with no detectable E. macusaniensis.

PROCEDURES: Alpacas were inoculated with 2 × 10(4) sporulated oocysts. Serial fecal samples collected during the next 38 days were tested via sucrose flotation and PCR assay.

RESULTS: Oocyst passage was detected via fecal flotation in all 4 alpacas 31 to 35 days after inoculation. Three had positive results for PCR assays on samples obtained 7 to 14 days after inoculation. One alpaca subsequently was removed from the study because of weight loss and inappetence. Two remaining alpacas had positive PCR reactions 28 and 31 days after inoculation, up to 7 days before oocysts appeared in the feces. All fecal samples with positive results for flotation also had positive results for PCR assay.

CONCLUSIONS AND CLINICAL RELEVANCE: The PCR assay was able to detect early (7 to 14 days) and late (28 to 31 days) prepatent infection. These positive results suggested that the assay could have been detecting DNA unassociated with oocysts or detecting shedding earlier than has been previously recognized. The gap between the early and late detection periods may not be evident in alpacas receiving a larger or continuous inoculum, as might occur with natural infection. Use of a PCR assay for analysis of fecal samples may be valuable for detection of E. macusaniensis during the prepatent period, thus aiding in the identification and control of infected animals.

DOI10.2460/ajvr.73.1.13
Alternate JournalAm. J. Vet. Res.