J Vet Sci.  2017 Dec;18(4):547-549. 10.4142/jvs.2017.18.4.547.

Surveyor assay to diagnose persistent Müllerian duct syndrome in Miniature Schnauzers

Affiliations
  • 1Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for Regenerative Medicine, Dankook University, Cheonan 31116, Korea. shim@dku.edu
  • 2Dream Animal Hospital, Seoul 05570, Korea.
  • 3Institute of Tissue Regeneration Engineering, Dankook University, Cheonan 31116, Korea.
  • 4Department of Physiology, Dankook University School of Medicine, Cheonan 31116, Korea.

Abstract

Persistent Müllerian duct syndrome (PMDS) is a pseudohermaphroditism in males characterized by the presence of Müllerian duct derivatives. As PMDS dogs often lack clinical symptoms, a molecular diagnosis is essential to identify the syndrome in these animals. In this study, a new molecular method using DNA mismatch-specific Surveyor nuclease was developed. The Surveyor nuclease assay identified the AMHR2 mutation that produced PMDS in a Miniature Schnauzer as accurately as that obtained by using the conventional method based on restriction digestion. As an alternative to the current molecular diagnostic method, the new method may result in increased accuracy when detecting PMDS.

Keyword

Miniature Schnauzer; persistent Müllerian duct syndrome; surveyor nuclease assay

MeSH Terms

Animals
Diagnosis
Digestion
Disorders of Sex Development
DNA
Dogs
Humans
Male
Methods
Pathology, Molecular
DNA

Figure

  • Fig. 1 Gross appearance of the uterus and testes surgically removed from a PMDS-affected Miniature Schnauzer. U, uterus; T, testis; V, vagina.

  • Fig. 2 (A) Restriction digestion of AMHR2 from PMDS and normal Miniature Schnauzers. The PMDS dog had 212, 333, and 43 bp DNA fragments, whereas the normal dog had 255, 333, and 74 bp DNA fragments. (B) Surveyor nuclease assay of AMHR2 from PMDS and normal Miniature Schnauzer. The 662 bp heteroduplexed PCR amplicon is cleaved into 451 bp and 211 bp fragments in the PMDS dog, whereas no cleavage was detected in the normal dog. The 451 bp-band restricted by Surveyor nuclease is indicated by an asterisk. (C) C241T mutation detection by direct sequencing of AMHR2 using the PCR product from the PMDS Miniature Schnauzer. The asterisk indicates a C to T conversion.


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