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用于通过DNA扩增检测卡氏肺孢子虫的口咽样本。

Oropharyngeal samples for detection of Pneumocystis carinii by DNA amplification.

作者信息

Wakefield A E, Miller R F, Guiver L A, Hopkin J M

机构信息

Department of Paediatrics, John Radcliffe Hospital, Oxford.

出版信息

Q J Med. 1993 Jun;86(6):401-6.

PMID:8171187
Abstract

Pneumocystis carinii pneumonia is a major complication of T-lymphocyte immune deficiency. Restriction of the disease to the alveolar spaces and failure to culture R. carinii has hindered simple diagnostic methods. We have developed a specific DNA amplification method for P. carinii and shown diagnostic sensitivity and specificity exceeding 95% for pneumocystis pneumonia when applied to bronchoscopic lavage and hypertonic saline induced sputum. We here report application of DNA amplification to simple oropharyngeal samples in 31 HIV-positive patients with respiratory illness. P. carinii-specific DNA was detected in 10 of 18 (56%) patients with pneumocystis pneumonia by ethidium bromide stained gels and 14 of 18 (78%) patients by the more sensitive technique of oligoblotting. P. carinii DNA was not detected in samples from 13 patients with other respiratory diagnoses. An oropharyngeal sample offers a simple specimen for detecting P. carinii by DNA amplification; refinements of technique and calibration may allow its development for accurate diagnostic and epidemiological work.

摘要

卡氏肺孢子虫肺炎是T淋巴细胞免疫缺陷的主要并发症。该疾病局限于肺泡腔且无法培养卡氏肺孢子虫,这阻碍了简单诊断方法的发展。我们开发了一种针对卡氏肺孢子虫的特异性DNA扩增方法,并且在应用于支气管灌洗和高渗盐水诱导痰液时,对肺孢子虫肺炎显示出超过95%的诊断敏感性和特异性。我们在此报告了DNA扩增在31例患有呼吸道疾病的HIV阳性患者的简单口咽样本中的应用。通过溴化乙锭染色凝胶,在18例肺孢子虫肺炎患者中的10例(56%)检测到卡氏肺孢子虫特异性DNA,通过更敏感的寡核苷酸印迹技术,在18例患者中的14例(78%)检测到。在13例患有其他呼吸道诊断疾病的患者样本中未检测到卡氏肺孢子虫DNA。口咽样本为通过DNA扩增检测卡氏肺孢子虫提供了一种简单的标本;技术的改进和校准可能使其发展用于准确的诊断和流行病学工作。

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