The use of PCR to detect Neospora caninum DNA in the blood of naturally infected cows

TitleThe use of PCR to detect Neospora caninum DNA in the blood of naturally infected cows
Publication TypeJournal Article
Year of Publication2004
AuthorsOkeoma, C.M., Williamson N.B., Pomroy W.E., Stowell K.M., and Gillespie L.
JournalVeterinary Parasitology
Volume122
Issue4
Pagination307 - 315
Date Published2004
ISBN Number03044017 (ISSN)
KeywordsAbortion, Veterinary, Animals, Antibodies, Protozoan, article, blood, blood analysis, blood sampling, cattle, Cattle Diseases, Coccidiosis, controlled study, cow, DNA, DNA determination, DNA fragment, DNA isolation, DNA Primers, DNA, Protozoan, female, Fluorescent Antibody Technique, Indirect, gene amplification, gene identification, gene sequence, gestational age, internal transcribed spacer 1, molecular cloning, Neospora, Neospora caninum, nonhuman, parasite identification, PCR, polymerase chain reaction, Pregnancy, protozoal DNA, sequence analysis
Abstract

Twelve 2-year old heifers in their fifth month of gestation when pregnancy tested were used in this study. Six heifers aborted at approximately 4 months of gestation and had blood samples drawn less than 6 weeks after the abortions were identified. Blood samples were also drawn from three sero-positive pregnant and three sero-negative pregnant heifers. DNA was isolated from the samples and a 350 bp fragment of the Nc-5 gene was PCR amplified using primer pair Np21+ and Np6+. Also, the Internal Transcribed Spacer 1 (ITS1) was PCR amplified using Tim 3 and Tim 11 primer pair. The Nc-5 gene fragment was cloned, sequenced and the sequence BLAST-tested. Similarly, the ITS1 product was sequenced and BLAST-tested. The BLAST test results revealed that Neospora caninum DNA was present in these blood samples indicating that polymerase chain reaction can be used in the detection of N. caninum DNA in the blood of sero-positive cows. © 2004 Elsevier B.V. All rights reserved.

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