消光型蛍光プローブを用いたリアルタイムＰＣＲ法による水中のクリプトスポリジウムの定量および種別判定手法の開発

DOI

書誌事項

タイトル別名
  • Development of Quantification and Genotyping methods for Cryptosporidium in water by Quenching Probe PCR followed by RFLP

説明

A new method was developed to quantify Cryptosporidium in water. Quenching Probe PCR (QProbe-PCR) technique could successfully amplify approximately 1280bp of Cryptosporidium 18S rDNA from a sample with as low as 60 [oocysts/tube] of Cryptosporidium parvum bovine genotype. QProbe-PCR showed high accuracy and high sensitivity compared to Real Time PCR with TaqMan probe.<BR>QProbe-PCRh as an advantaget hat the PCR products can be applied for molecular characterization. Arestriction fragment length polymorphism (RFLP) technique was used to distinguish Cryptosporidium species and genotypes. Five species (C. parvum bovine genotype, C. parvum human genotype, C. meleagridis, C. felis and C. muris) could be distinguished by the RFLP with restriction enzymes Ssp I, Vsp I and Sty I. The Sty I successfully differentiated C. muris calf genotype (also known as C. andersoni) and C. muris mouse genotype. Database-based analysis revealed that 8 species out of 10 could be distinguished by RFLP with these three restriction enzymes.<BR>QProbe-PCR-RFLP techniques can provide information on the genotype as well as the quantity of Cryptosporidium from the same sample. This technique can be a useful tool for waterborne risk assessment of Cryptosporidiosis.

収録刊行物

キーワード

詳細情報 詳細情報について

  • CRID
    1390282679089503232
  • NII論文ID
    130003949381
  • DOI
    10.11532/proes1992.41.311
  • ISSN
    1884829X
    13415115
  • 本文言語コード
    ja
  • データソース種別
    • JaLC
    • CiNii Articles
  • 抄録ライセンスフラグ
    使用不可

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