A reproducible assay of polymerase chain reaction to detect trinucleotide repeat expansion of Huntington's disease and senile chorea

Mitsunori Watanabe, Koji Abe, Masashi Aoki, Takeshi Kameya, Yasuto Itoyama, Mikio Shoji, Masaki Ikeda, Tomomichi Iizuka, Shunsaku Hirai

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

A simple and reproducible method of polymerase chain reaction (PCR) assay was established to detect trinucleotide repeat expansion for Huntington's disease (HD) using a new DNA polymerase and buffer system. The system consists of an extremely heat stable DNA polymerase (Pfu), and a buffer supplemented with ammonium sulfate and dimethyl sulfoxide. Previous methods to amplify expanded alleles for HD have been very complex in PCR conditions, but the reproducibility was sometimes very low because of repetitive sequences around the primer sequences. With the present method, strong bands for the disease alleles were reproducibly visible in a conventional agarose gel stained with ethidium bromide without using isotopes. Three cases with sporadic HD and a case with senile chorea showed expanded alleles for HD with smaller sizes of the expansion than cases with typical HD. These results showed that the present method provides a simple and reproducible way to detect HD allele, and some cases with sporadic HD and senile chorea had expanded HD alleles.

Original languageEnglish
Pages (from-to)16-18
Number of pages3
JournalNeurological Research
Volume18
Issue number1
DOIs
Publication statusPublished - 1996

Keywords

  • Huntington's disease
  • Pfu DNA polymerase
  • nonradioactive
  • polymerase chain reaction
  • senile chorea

ASJC Scopus subject areas

  • Neurology
  • Clinical Neurology

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