寡核苷酸探针RNA原位杂交法检测肺孢子虫的研究

Detection of Pneumocystis carinii on the lung paraffin section by RNA in situ hybridization

目的探讨RNA原位杂交法在肺孢子虫病理检测中的应用。方法采用小亚单位RNA位点来源的寡核苷酸探针、地高辛加尾标记、Wistar雌性大鼠皮下注射地塞米松建立肺孢子虫肺炎动物模型,取肺组织制备石蜡标本进行RNA原位杂交,并将结果与瑞氏-吉姆萨染色法进行比较。结果 RNA原位杂交法于感染后第3周检到虫体,呈游离分布;7～8周时检测到大量包囊;9～10周时滋养体大量出现;组织内肺孢子虫的检出率(32/32)高于瑞氏-吉姆萨染色法(25/32)。结论 RNA原位杂交法是一种敏感特异的肺孢子虫病理检测方法。

Objective To investigate the possible application of RNA in situ hybridization for the detection of Pneumocystis carinii. Methods The Pneumocystis carinii infected Wistar rat model was established. Rats inoculated with Pneumocystis carinii were sacrificed 3, 4, 5, 6, 7, 8, 9 and 10 weeks after injection with Dexanethasone, and the paraffin sections of lung were prepared to perform the in situ hybridization with Dig-labeled oligonucleotide probe complememtary to small subunit rRNA of Pneumocystis carinii. Results It was found that the specific positive results were observed on the paraffin sections 3 weeks after injection.All the 32 paraffin sections detected by in situ hybridization demonstrated positive results,while only 25 sections showed positive results by Wright-Giemsa＇s staining. Conclusion The RNA in situ hybridization is a sensitive and specific method for the detection of Pneumocystis carinii in pathological sections.