PCRによる犬の尿中レプトスピラ検出法の検討および臨床例への応用  [in Japanese] PCR for Detecting Leptospira in Canine Urine and Its Clinical Applications  [in Japanese]

Access this Article

Author(s)

Abstract

犬の尿中レプトスピラの検出感度を, First PCRとBooster PCRについて比較した.<I>Leptospira</I> icterohaemorrhagiaeを混入した蒸留水および犬の尿から, 加熱処理により抽出したDNAサンプルを用いてFirst PCRを, またFirst PCRで増幅されたPCR産物をDNAサンプルとして100倍に希釈し, 12,000rpmで2分間遠沈後Booster PCRを行った. 検出された最少菌数は, First PCRでは蒸留水中では1.6×10<SUP>3</SUP>個/m<I>l</I>, 尿中では1.6×10<SUP>4</SUP>個/m<I>l</I>であったが, Booster PCRでは蒸留水中および尿中ともに1.6×10個/m<I>l</I>であり, 検出感度が上昇した. 臨床例において, 発症後1週間以内の尿で両PCR法により本菌の存在が認められ, 従来の説よりも早期にレプトスピラが尿中に排出されることが示唆された. PCRによる尿中レプトスピラの検出が早期診断に有用であることが示された.

A comparative study was made for evaluating the detection sensitivity of first PCR and booster PCR for <I>Lep-tospira</I> in canine urine. Heat treatment was used to extract DNA samples from distilled water and canine urine, both containing <I>Leptospira</I> icterohaemorrhagiae. These samples were then submitted to first PCR. In addition, PCR products amplified by first PCR were used as DNA samples that were diluted (1: 100), centrifuged at 12, 000 rpm, and submitted to booster PCR. In first PCR, minimum numbers of <I>Leptospira</I> were 1.6×10<SUP>3</SUP>/m<I>l</I> in distilled water and 1.6×10<SUP>4</SUP>/m<I>l</I> in canine urine. In booster PCR, however, the figure was 1.6×10/m<I>l</I> for both distilled water and urine. This suggests that booster PCR is more sensitive than first PCR. In clinical cases, both PCR assays detected <I>Leptospira</I> in canine urine within one week after manifestation of signs sug-gestive of leptospirosis. This indicates that <I>Leptospira</I> appear in the urine earlier in the course of the disease than had previously been thought. PCR assays of <I>Leptospira</I> in canine urine were shown to be useful in the early diagnosis of leptospirosis.-Key words: antibody, dog, Lettos/ira, PCR, urine.

Journal

  • Journal of the Japan Veterinary Medical Association

    Journal of the Japan Veterinary Medical Association 54(4), 288-292, 2001-04-20

    Japan Veterinary Medical Association

References:  17

Cited by:  1

Codes

  • NII Article ID (NAID)
    10008760689
  • NII NACSIS-CAT ID (NCID)
    AN00191857
  • Text Lang
    JPN
  • Article Type
    Journal Article
  • ISSN
    04466454
  • NDL Article ID
    5754874
  • NDL Source Classification
    ZR22(科学技術--農林水産--畜産)
  • NDL Call No.
    Z18-352
  • Data Source
    CJP  CJPref  NDL  J-STAGE  JASI 
Page Top