原位杂交技术鉴定马尔尼菲青霉

Identification of Penicillium Marneffei by in Situ Hybridization Technique

目的 探讨原位杂交技术 (ISH)特异性检测马尔尼菲青霉 (Pm)感染组织标本的可行性。方法 以Pm的rDNA区间种特异性序列作为特异性探针 ,通过聚合酶链反应 (PCR)扩增合成并以地高辛标记 ,与 12例临床确诊的Pm感染组织标本中的靶DNA杂交。结果 12例Pm感染标本中全部阳性 ,其它种属真菌感染组织均为阴性。结论 ISH技术可以灵敏、特异地检测出组织中的Pm ,适用于今后临床组织标本中Pm感染的检测。

Objective To evaluate the applicability of in situ hybridization (ISH) technique in the specific diagnosis of clinical tissue specimens infected with penicillium marneffei(Pm).Methods Twelve formalin-fixed and paraffin-embeded tissue specimens from patients with Pm infected were studied by an ISH technique, The specific probe was selected from a Pm rDNA region, synthesized and digoxigenin labeled by means of polymerase chain reaction (PCR).Results Positive staining of Pm was observed in all 12 tissue specimens infected with Pm,but none of positive staining occurring in other genus or species fungi had been found.Conclusion The ISH is a sensitive and specific technique for identification of Pm infection, It could be applicability to clinical molecular diagnosis of Pm infection.