中华皮肤科杂志 ›› 2009, Vol. 42 ›› Issue (9): 622-624.

• 论著 • 上一篇    下一篇

以S2-R2为引物PCR检测石蜡切片中的孢子丝菌

王东1,刘晓明2,张振颖3   

  1. 1. 沈阳市第七人民医院
    2. 大连医科大学附属第一医院皮肤科
    3. 大连医科大学附属第一医院皮肤科教研室
  • 收稿日期:2008-09-08 修回日期:2009-02-05 出版日期:2009-09-15 发布日期:2012-03-21
  • 通讯作者: 王东 E-mail:wangdong19820210@126.com
  • 作者简介:
  • 基金资助:
    国家自然科学基金资助项目(编号)

Detection of Sporothrix schenckii in paraffin-embedded tissue with a primer pair S2-R2

  • Received:2008-09-08 Revised:2009-02-05 Online:2009-09-15 Published:2012-03-21

摘要: 目的 探讨应用特异性引物PCR方法检测石蜡切片中孢子丝菌的可行性。方法 选取30份大连及10份长春地区临床疑诊孢子丝菌病的石蜡切片标本,采用改良的微波脱蜡、液氮研磨-CTAB破壁法提取DNA,以特异性引物S2-R2进行PCR扩增,与真菌培养结果进行比对。结果 30份大连地区标本中22份见阳性PCR扩增产物(73.33%)。真菌培养阳性的22份标本中20份PCR出现阳性扩增产物(91%),真菌培养阴性的8份标本中2 份PCR出现阳性扩增产物。10份长春地区标本7份见阳性PCR扩增产物(70%)。结论 以S2-R2为引物的 PCR适用于孢子丝菌病石蜡切片中病原菌的检测。

关键词: 孢子丝菌病, 孢子丝菌属, DNA引物

Abstract: Objective To explore the feasibility of detecting Sporothrix schenckii (S. schenckiii) in paraffin-embedded tissue by polymerase chain reaction (PCR) using a specific primer. Methods A total of 30 paraffin-embedded tissue samples were collected from patients with suspected sporotrichosis in Dalian and 10 samples from those in Changchun. DNA was extracted from these samples via modified microwave- based deparaffinage and gringding in liquid nitrogen and incubation with CTAB. PCR was performed with a specific primer pair S2-R2 targeting the chitin synthase 1 gene of S. schenckii, and the results of PCR were compared with those of culture. Results Expected DNA fragments of S. schenckii were amplified from 22 (73.33%) of 30 samples from Dalian and 7 (70%) of 10 samples from Changchun. Characteristic colony of S. schenckii grew from 22 samples from Dalian, of which, 20 were positive for PCR, while in the remaining 8 samples negative for culture, only 2 samples were positive for PCR. Conclusion PCR with the specific primer pair S2-R2 can be used to detect S. schenckii in paraffin-embedded tissue.