Real-time PCR for sexing Schistosoma mansoni cercariae

Frédéric D. Chevalier, Winka Le Clec'h, Ana Carolina Alves De Mattos, Philip T. Loverde, Timothy J.C. Anderson

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

The gender of cercarial larvae can only be determined using molecular methods. End point PCR methods that amplify repetitive markers on the W chromosome of the female (ZW) parasites have been developed, but sometimes results are ambiguous or incorrect. To more effectively distinguish sexes, and to determine why end point PCR can be incorrect, we quantified the W6 repeat sequence and a specific Z chromosome gene using real-time PCR. The ratio between copy number of W6 and a Z chromosome marker unambiguously identifies gender: females have higher ratios (421-4371) than males (0-21). However, some males have low numbers of W6 elements in their genome, and qPCR demonstrated significantly higher W6/Z marker ratios for male genotypes giving ambiguous end point PCR results compared with males giving clear end point results. The quantitative PCR sexing method developed will be particularly useful where reliable sexing of cercariae is critical, for example when staging genetic crosses.

Original languageEnglish (US)
Pages (from-to)35-38
Number of pages4
JournalMolecular and Biochemical Parasitology
Volume205
Issue number1-2
DOIs
StatePublished - Jan 1 2016

Keywords

  • Cercariae
  • Schistosome
  • Sexing methods
  • qPCR

ASJC Scopus subject areas

  • Parasitology
  • Molecular Biology

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    Chevalier, F. D., Le Clec'h, W., Alves De Mattos, A. C., Loverde, P. T., & Anderson, T. J. C. (2016). Real-time PCR for sexing Schistosoma mansoni cercariae. Molecular and Biochemical Parasitology, 205(1-2), 35-38. https://doi.org/10.1016/j.molbiopara.2016.03.010