Identification of bovine Neospora parasites by PCR amplification and specific small-subunit rRNA sequence probe hybridization

S. Y H O Michael, Bradd C. Barr, Antoinette E. Marsh, Mark L Anderson, Joan D Rowe, Alice F Tarantal, Andrew G Hendrickx, Karen Sverlow, J. P. Dubey, Patricia A Conrad

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

Neospora is a newly recognized genus of pathogenic coccidia, closely related to Toxoplasma gondii, that can cause abortion or congenital disease in a variety of domestic animal hosts. On the basis of the small-subunit rRNA gene sequences of Neospora spp. and other apicomplexa coccidia, oligonucleotide primers COC-1 and COC-2 were used for PCR amplification of conserved sequences of approximately 300 bp in size. A Neospora-specific chemiluminescent probe hybridized to Southern blots of amplification products from Neospora DNA but not to Southern blots with amplified DNA from the other coccidian parasites tested. A Toxoplasma-specific probe whose sequence differed from that of the probe for Neospora spp. by a single base pair was used to distinguish these parasites by specific Southern blot hybridization. The PCR system detected as few as one Neospora tachyzoite in the culture medium or five tachyzoites in samples of whole blond or amniotic fluid spiked with Neospora parasites. In addition, Neospora PCR products were successfully amplified from whole blood and amniotic fluid samples of experimentally infected bovine and rhesus macaque fetuses. These results indicate that this PCR and probe hybridization system could be a valuable adjunct to serology and immunohistochemistry for the diagnosis of Neospora infections in bovine or primate fetuses.

Original languageEnglish (US)
Pages (from-to)1203-1208
Number of pages6
JournalJournal of Clinical Microbiology
Volume34
Issue number5
StatePublished - 1996

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Neospora
Parasites
Polymerase Chain Reaction
Southern Blotting
Coccidia
Toxoplasma
Amniotic Fluid
Fetus
Apicomplexa
DNA Primers
Conserved Sequence
DNA
Domestic Animals
Serology
Macaca mulatta
rRNA Genes
Base Pairing
Primates
Culture Media
Immunohistochemistry

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Identification of bovine Neospora parasites by PCR amplification and specific small-subunit rRNA sequence probe hybridization. / Michael, S. Y H O; Barr, Bradd C.; Marsh, Antoinette E.; Anderson, Mark L; Rowe, Joan D; Tarantal, Alice F; Hendrickx, Andrew G; Sverlow, Karen; Dubey, J. P.; Conrad, Patricia A.

In: Journal of Clinical Microbiology, Vol. 34, No. 5, 1996, p. 1203-1208.

Research output: Contribution to journalArticle

Michael, S. Y H O ; Barr, Bradd C. ; Marsh, Antoinette E. ; Anderson, Mark L ; Rowe, Joan D ; Tarantal, Alice F ; Hendrickx, Andrew G ; Sverlow, Karen ; Dubey, J. P. ; Conrad, Patricia A. / Identification of bovine Neospora parasites by PCR amplification and specific small-subunit rRNA sequence probe hybridization. In: Journal of Clinical Microbiology. 1996 ; Vol. 34, No. 5. pp. 1203-1208.
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