爪白癬からの起因菌同定における培養法とPCR-RFLP法の比較検討  [in Japanese] Comparative Study between Culture and PCR-RFLP Analysis on Identification of the Causative Agent of Tinea Unguium  [in Japanese]

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Abstract

当科外来を受診した爪白癬患者100例から得られた罹患爪の一部をサブローブドウ糖寒天培地に培養し, 27℃にて, 最長2ヵ月間観察した。残りの爪サンプルを凍結, 超音波破砕しDNAを抽出, リボゾームDNA上のITS領域を用いたPCRをおこない, 得られたPCR産物を<i>Mva</i> I, <i>Hin</i> f Iの各制限酵素で消化し, 切断パターンを分析した. また, 爪サンプルの重量を測定し, PCR-RFLP法の結果と比較した. 結果は培養法の同定率が20%にとどまったのに対し, PCR-RFLP法では73%であった. 爪サンプルの重量とPCR-RFLP法での同定結果との間には有意差は認められなかった. サブローブドウ糖培地を用いた培養による爪白癬の起因菌同定に際し, 菌のviabilityの問題から培養成功率の低さが指摘されていたが, 爪サンプルから直接行うPCR-RFLP法は迅速かつ成功率の高い同定法であると思われる.

Background: To identify the pathogenic fungi of dermatophytosis, restriction fragment length polymorphism (RFLP) analysis of PCR amplified ribosomal DNA including internal transcribed spacers (ITS) has been established in Japan. Our purpose was to evaluate the usability of PCR-RFLP analysis to identify the causative agent of tinea unguium directly from a nail sample.<br>Method: Samples of tinea unguium from 100 nails were collected and cultured on Sabouraud's glucose agar and observed for 2 months. DNA was extracted from these samples, and the PCR product was digested with restriction enzymes <i>Mva</i> I and <i>Hin</i>f I. Weight of the samples was determined.<br>Result: Sensitivity of PCR-RFLP analysis (73%) was higher than that of culture (20%) showing that PCR is more advantageous for identification of the causative agent of tinea unguium. Sensitivity of PCR-RFLP did not depend on weight of the nail sample.

Journal

  • Nippon Ishinkin Gakkai Zasshi

    Nippon Ishinkin Gakkai Zasshi 47(1), 11-14, 2006-01-30

    The Japanese Society for Medical Mycology

References:  11

Cited by:  3

Codes

  • NII Article ID (NAID)
    10017242449
  • NII NACSIS-CAT ID (NCID)
    AN10166867
  • Text Lang
    JPN
  • Article Type
    Journal Article
  • ISSN
    09164804
  • NDL Article ID
    023253058
  • NDL Call No.
    Z19-348
  • Data Source
    CJP  CJPref  NDL  J-STAGE 
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